UNIT 18 METAMORPHOSIS

18.1 Introduction
Objectives
18.2 Types of development
18.3 Types of metamorphic changes
18.4 Larval forms in various animal groups
Significance of ecological factors on the evolution, utility and distribution of larval
forms
A catalogue of larval forms in various animal groups
18.5 Metamorphosis in Amphibians
The process of metamorphosis in anurans
The process of metamorphosis in urodeles
Hormones in metamorphosis of amphibia
Interaction of amphibian hormones in the process of metamorphosis
Tissue reactivity
Induction in metamorphosis
Molecular response to thyroid hormones during metamorphosis
Neoteny
18.6 Development, growth and metamorphosis in insects
General process of post-hatching growth in insects
Patterns of metamorphosis
Factors controlling metamorphosis in insects
Organs and hormones involved in insect metamorphosis
Interaction of insect hormones in the process of metamorphosis
Effect of metamorphic hormones on gene expression in moulting and
metamorphosing insects
18.7 Comparison between metamorphosis in amphibians and insects
18.8 Summary
18.9 Terminal Questions
18.10 Answers

18.1 INTRODUCTION
In the earlier units of this course you have read that most animals develop directly,
their zygote undergo multiplication, differentiation and morphogenesis to become a
young one, similar to the adult, except in size and sexual maturity. However, in many
animals groups ranging from the Porifera to the vertebrates deve.lopment from
zygote to adult is often indirect. These animals have one or several intervening
larval stages in their developmental cycle before they develop into an adult by a
sudden dramatic transformation known as 'Metamorphosis'.

The phenomenon of metamorphosis is defined as a 'process during development

which involves a dramatic change in morphology and physiology of the larva, so
that it is transformed into an adult with completely different morphology and
physiology, often for life in a different habitat.
In many such animals undergoing metamorphosis, the larva is usually very different
from the adult. The best known examples are tadpoles of frogs, caterpillar of
butterflies and moths, tadpole larva of ascidians, various larval types of
crustaceans, ciliated uochophores of marine annelids and molluscs etc; In fact s o w
times the difference between the larva and adult is so great that without knowing the
origin of the egg, or without following the young one through its full development, it
would be next to impossible to know that the young and the adult are of the same
species. In the past such differences have sometimes led to the larva and adult of
the same species being assigned to different taxonomic groups. For example the
larva and adult of the axolotl Ambystoma (Urodele--amphibia) and Tribegulians of
blister beetle (Inaecta) till quite recently were mistakenly assigned to different
species. &re than a century ago, only the study of metamorphosis of the tadpole
lawa of ascidians could decide that the ascidians belong to Phylum Chordata.

The changes that occur due to metamorphosis relate often to a change in habitat with
a corresponding change in the organisms' suucture and other features. For example,
in sea urchins, there occurs a change from planktonic to benthic existence, in frogs
and toads from an aquatic to a terrestrial m'bde of life and in insects from a non-flying -
to a flying life.
In the present unit, which deals with metamorphosis, you will be studying this
phenomenon in two diverse animal groups: namely amphibians among the
vertebrates and insects among the invertebrates.
Your study will show that the metamorphic process in animals differ in the
nature of transformation and in the mode of causation, making it impossible to
describe metamorphosis in a generalized manner; however it must be pointed out
that there are certain basic similarities.
In all these animals, development does not stop at hatching but continues during
post embryonic life, and the principles that apply to embryonic development also
apply to post embryonic development. Furthermore, the post development process
in such animals are usually found to be reactivated by specific hormones, which
modify and modulate the timing and duration of the process.

The metamorphic changes involve differential desuuction of certain tissues

'

accompanied by an increase in growth and differentiation of other tissues.

Objectives
After studying this unit, you will be able to:
describe the types of developments and metamorphosis in animals
define the terms metamorphosis, ametabolous, hemimetabolous, holometabolous.
moult, instar, nymph, naiad, stadium, pupa, imago and diapause
enumerate tbe various larval f o q s occurring in different animal groups
\ '
explain the process of amphibian and insect metamorphosis and its conuol by
hormones
explain the interactions of the various hormones which cause metttmorphosis in
amphibians and insects
describe the effect of metamorphic hormones on gene expression in
amphibians and insects

TYPES OF DEVELOPMENT
Different animals have evolvcd different methods of development. These
methods can be broadly classified into two categories (i) direct development (ii)
indirect development.
 Direct development
In some animals whose eggs have little or no yolk, such as the placental mammals,
the embryos develop and grow within the womb of the mother from which they
also obtain nourishment. Once, the offspring has developed to a stage where it
looks like a miniature adult, the mother gives binh to it. This young one
achieves its adult size and sexual maturity after birth over a period of time by
gradual growth.
In animals whose eggs have a large amount of yolk. such as those of the birds and
reptiles, the eggs develop oviparously outside the mother's body. Young ones
resembling the miniature adults hatch out. The young individual then attains
adult size and sexual maturity by gradual growth over a length of time.
Indirect development
In many animals like frog. Amphioxus, Herdmania, insects and many other
invertebrates and vertebrates, the offspring which hatches out from the egg looks
very different from the adult form, and is called a larva. The larva leads an
independent existence for some time that varies from species to species, and
then transforms into a miniature adult by the process of metamorphosis.

18.3 TYPES OF METAMORPHIC CHANGES

The process of metamorphosis involves reactivation of the morphogenetic
processes. The morphogenetic changes as well as the mode of causation of these
changes vary in different animal groups. The degree of changes that occur
during metamorphosis depend on the degree of difference between the larva and the
adult forms. For example, in urodele amphibians (newts) and
hemimetabolous insects (cockroach), larva and adult show a few differences. The
metamorphic changes are relatively less and metamorphosis is said to be gradual
or incomplete. On the other hand, in anuran amphibians (frogs) and
holometabolous insects, where differences between the larva and adult are
enormous, the changes during metamorphosis are extensive and drastic. This type
of metamorphosis is called radical or complete metamorphosis.
The changes during metamorphosis, include those of structural, physiological and
biochemical nature. These are marked by disintegration and atrophy of some
structures, cellular death in some tissues, morphogenesis and differentiation of
certain new structures and remodelling of some others. These changes of
metamorphosis in at least some groups of animals (insects, crustaceans,
amphibians), are known to be controlled by hormones which serve as the causative
agents of metamorphosis.
, SAQ 1
Explain the term (i) direct development (ii) indirect development.

18.4 LARVAL FORMS IN VARIOUS ANIMAL GROUPS

18.4.1 Significance of ecological factors on the evolution, utility

and distribution of larval forms
Environmental factors surround and influence the egg and the young throughout the
Animal ~wdoprnent animal kingdom. These factors affect such vital activities as the place where the eggs
are laid, the amount of food provided for the young during early development,
the various devices acquired for their protection, the specialized modifications for
accomplishing locomotion and the production of those larvae which change into
adult by metamorphosis.
The larvae which undergo metamorphosis present a wide range of variations as
I

compared to the animals which undergo direct development. You are already
aware that usually in animals which undergo metainorphosis the larvae and adult live
in different habitats and consequently in different environments. Metamorphosis
appears essential for those forms in which food and I habits of the adult are
unsuitable for the developing young.
Among the three habitats, namely saline water, fresh water and land, most of the
animals which undergo metamorphosis are found in *e marine waters. This seems to
be in line with the assumption based on evolutionarb evidence that all organisms
originated in the sea and that many types of aninhals develop in a moist
atmosphere or in liquid medium. Futhermore, the transition from saline to fresh
water, or to terrestrial life present great difficulty to the developing young, and so
those animals which live in fresh water or land are Fnore often characterized by
direct development.
The sea medium also provides a more constant environment: the temperature rarely
fluctuates, the sea water rarely dries up or freezes, food is available aplenty at the
surface where most of the metamorphic forms occur.

Metamorphosis is rarely seen in animals occurring in fresh water. This is because

the living conditions of the fresh water environment lacks constancy as compared
to that of the sea. Except in a few cases the fresh water bodies usually lack
-., sufficient depth and area to maintain even a minimum degree of stability. Fresh
water heats and freezes much more rapidly than the sea, and irs stream floods and
recedes. leaving many forms of life to perish. Furthermore, every stream has depths
and narrows where flow is rapid, and flats where flow is slow, as a result of which
the vulnerable larvae find it difficult to maintain themselves. These are some of
the reasons as to why most animals in fresh water have direct development, where the
young and adult are similar and share a similar environment.

The terrestrial environment appears to be the most inhospitable for the

developing larva with fluctuations in temperature, glare of the light, lesser moisture
and easy exposure to enemies. It is due to these reasons that in land forms
development is either direct and fairly simple or greatly specialized with
complicated adaptations and structures for protecting the young during the
immature period. The latter type of development is exemplified by many forms of
insects in which the life cycle is complicated and metamorphosis occurs.

These different environmental factors and habigts are responsible for the lack of
uniformity in the occurrence of larval types in the different groups of animal kingdom.
The particular forms of larvae within a group are of adaptive rather than
taxonomic significance. For example, in those animals hose adults are sessile the
young are active. This activity has two obvious reaso; first the species secures
its dispersal through migration of the young, and second, it extends the range.
This migration is commonly passive but over a period of time it successfully
serves to extend the range of the species, as if the adults themselves were able to
move. Furthermore, to ensure disperal large amount of gametes are produced, as
the chance method of fertilization results in the loss of many sperms and eggs.
Also, all fertilized eggs do not reach maturity as larval mortality is high due to
various physical factors, predators etc. Another example of adaption is observed in
the marine lobster which lives at the bottom region, is negatively phototropic and
feeds on whatever comes its way, unlike irs larvae which are positively
phototropic and live at the surface of the sea where food is abundantly available. In
such a life style it has various structural modifications which change after
metamorphosis into an adult. Similarly, other animals have specialized structures
for their survival suitable to their environment and needs. You should keep in
mind, though, that the larvae would survive only if subsequent changes by
metamorphosis allow the organism to undertake a new mode of life. In other
words, metamorphosis id a necessity for bringing about those adaptations which
enable the organism to live in its permanent environment and reproduce Metamorphosis
successfully.
18.4.2 A catalogue of larval forms in various animal groups
The larval stages and types found in different groups are many. Each larval type
has a different structure and is known by a different name. Table 18.1 lists out the
names of some of the commonly studied larvae which occur in different
animal groups.

Table 18.1 : Examples of larval forms in major animal groups.

S.No. Animal group Larval form

Lower Mesozoa
Sponges amphiblastula (18.la)
Coelentrates planula (18.1b). actinula,
syphistoma, ephyra
Protosomia
Flat worms
Turbellarians miiiiFis larva
Trematodes miricidium, cercaria,
redia, sporocyst
Cestodes onchosphere
Nemertines pilidium
Annelids trochophore (18.2a)
Molluscs trochophore, veliger
Crustaceans nauplius, metanauplius
cypris, zoea, mysis, megalops
Insects
with incomplete metamorphosis Nymphs (18.2b) naiads
with complete metamorphosis maggots, caterpillars,
grubs (18.2b) pupae etc.
Ectoprocts Cyphonauts
Phoronids Actinauoch
Deurostomia
Echinoderms bipinnaria,
pluteus (18.3b).
18.3a) auricula
Hemichordates tornaria (18.3~)
Tunicates tadpole (18.3d)
Cyclostomes ammocoetes (18.3e)
True eels leptocephalus
Salmon alevin, pan, Smolt
Amphibians eft, tadpole
Animal Devdopment A look at the table will make you aware that metamorphosis is widespread in the
animal kingdom. It occurs in many phyla in the invertebrates, ranging from the
sponges (porifera) through the echinoderm and in several chordates. Fig. 18.1, 18.2
and 18.3 show some important larval forms which are often the subject of study.
However, in this unit too, we are not going to describe all the larval f&ms or their
metamorphosis. Instead we will restrict our study only to those aspects of larval life
and metamorphosis, that have been extensively studied by biologists. This unit is
devoted to the study of metamorphosis in amphibians and insects and will expose you
to some of the recent developments in the area of study.

Planula larva
Flagellated WUs

Granular WUB
01
eon-flagellated
WUS (b) i

Mouth I

Reproductive polyp

Vfp.18.1. LA,.. ~ of lower mesozoa

d f o ms
a) r AmphiL!estuia larva of porifera (ii) Adult sponge
b) (1) PPlanlila larva of a hydrozoan coeleatr~lv(il) Adult UBe&

.- - ... -- . -.- r::;?t$q7illar

- Iarvs
- <
. r. L
.
.
,. (^... ... <-" --.. 1
:) ,bJ

Fig $ 8 ",arts*: 'ax ,rs of scp- .+t?,l?aP * p naca;lr-hlr

a* (I I ruehopore I? J or dinn $id :ii: Advanced rogd:, mnrdmurphm!?
bj (i! Nymphs of rmetaborous inse.:ts ( i ~ )Nymphs (J Hemiraetabolous loser.@
t i l l L a&vae o f Holometabaou~idlsecrs
 @.. '3 . y$-y .:...,
.:
.... . .
.

development of a pluteus kna

Norocbord
of ophiumidm cchiwidea
Tailed larva (dl Attached larva dth rerorbcd mil Adult & d m
I
Ammocoete larva of lamprey

Tomaria larva
during rnetatnorpbosis
Toda larva.
I before rnetarn*hoI
Adult

- ..tunnel
um
Homy tooth
Young balaoglossus wonn
with tbree pairs of gill dita
(el

Fig.183: Larval forms of Deurostomea at various stages of metamorphosis

a. Blptnnarh Larva of Asterolden b. Pluteua larva of Echlnoldea c. Tornarln larva of Bnlanoglossus
d. Tadpde Larva of AscldIa 8. Ammomete larva of lampreJ
I
SAQ 2
i) State wbetber the following statements are true or false
(a) In animals which undergo metamorphosis the larvae and adult live in same
habitats and subsequently in same environments. TruePalse
(b) Metamorphosis is more commonly observed among the fresh water and
terrestrial organisms as compared to marine organisms. Truepalse
(c) The terrestrial environment is highly inhospitable for the developing larva.
TruePalse
(d) Metamorphosis essentially brings about those adaptations in organisms that
enable them to live in their permanent environment and reproduces successfully.
TrueFalse
(e) In terrestrial environment insects are the only group that pursue a free larval
life. Truepaise
Animal Development (f) Among the various animal groups metamorphosis has been studied in detail in
insects and amphibians. TrueFalse
ii) Match the following larvae with their adult groups.
a) Trochophore i) Cyclostomes
b) Tornaria ii) Crustacea
c) Bippinnaria iii) Echinoderms
d) Veliger iv) Balanoglossus
e) Zoea v) Annelids
f) Ammocoete vi) Molluscs

18.5 METAMORPHOSIS IN AMPHIBIANS

Metamorphosis is radical in anurans, slight or absent in urodeles. In anuran

amphibians like toads and most frogs, metamorphosis is usually associated with a
transition from an aquatic to a terrestrial or amphibious mode of life. Occasionally
however no transition in mode of life occurs as observed in the larval and adult of the
frog Xenopus laevis and many primitive anurans which remain aquatic throughout
their life. The change in habitat in the frogs and toads also usually results in a change
in their feeding habit. In some like Xlaevis there is no change in food habit since
both larvae and adult ale carnivorous.
Some anurans undergo an abbreviated type of mefamorphosis before hatching, as they
pass through a tailed, gilled tadpole-like stage within the jelly membrane of the egg.
Others undergo direct development by skipping the larval stages totally.
Metamorphosis in urodele amphibians is usually less striking. Some of them
undergo direct development, while others fail to complete their metamorphosis.
The. latter achieve sexual maturity as larvae, as seen in the axolotl larvae of
Ambystoma. This phenomenon is called neoteny, and will be discussed in section
185.8. Some urodeles like salamanders have been observed to undergo two
metamorphosis.
Metamorphosis in both anurans and urodeles essentially involves the activation of the
genomic set underlying the adult organization, which requires for its expression a
minimum mass of tissue that is greater than that of the egg.
The activation is believed to be due to the secretion .of a brain hormone which initiates
metamorphosis. The hormone triggers the degeneration of redundant larval organs and
growth of hitherto quiescent structures which are needed in the adult.
In amphibians the process of destruction and growth are smoothly coordinated, as a
result of which the animal retains its functional integrity throughout metamorphosis
instead of lying dormant as in the case of insects.

18.5.1 The process of metamorphosis in anurans

Metamorphosis is most dramatic in the anurans. In them it represents an intensive
period of growth and developmental changes during which the suucture and function
of almost every organ in the body is radically modified for adaptation to a new way of
life. Unlike other adaptational changes, however, this process is begun and normally
completed in the anticipation of the change of the environment. Some changes seen
during this process involve growth and maturation of the tissues concerned. Other
changes result in the regression and death of organs; the gills and tails which become
redundant in adult life are completely reabsorbed.The most familiar kinds of anurans
are frogs whose eggs develop into Ladpoles, which are limbless, aquatic creatures with
an oval torso and long, finned tail (Fig. 18.4). Metamorphosis transforms these
tadpoles into four-legged, hopping carnivorous adults (Fig.18.4). After metamorphosis
most types of anurans lead a terreslrial existence. returning to the water only to breed.
Toads also exhibit this type of life habit.
Tadpoles have many anatomical and physiological adaptations that distinguish them
from the adult. The fish like tadpole is herbivorous, as a result of which its mouth has
 two horny beaks with rows of horny teeth (Fig. 18.4 ) which help in rasping away plant
tissues. On each side of the head a fold of skin, the operculum, is present which covers
the gill that grow from the lower ends of the visceral arches. Lungs are rudimenlary
and nonfunctional. The epidermis has large pigment cells of different kinds and is
, underlaid by a jelly-like dermis rich in hyaluronic acid. Because of its herbivorous diet
the intestine of the tadpole is long and coiled. The eyes are r&ssed in the head, and
I the visual pigment porphyropsin is present in the retina. Nitrogenous waste products are
I
excreted primarily as ammonia. The red blood cells are produced mainly in the kidney
and the haemoglobin (HbF) present in the tadpoles is different from that of adult (HbA).
In order to transform into adults these tadpoles undergo three distinct phases of
metamorphic changes which are visible morphologically, and have been described by
Etkin as: (i) Premetamorphosis is defined as an initial period of extensive growth but
little developmental change. This is followed by (ii) Prometamorphosis during which
growth continues but conspicuous developmental changes such as the growth of hind
legs also take place. (Fig. 18.4b) Prometamorphosis ends with the emergence of the
forelimbs (Fig. 18.4~)and then the third and final stage of development, (iii) the
Metamorphic climax sets in. This is a comparative brief period in which profound
morphological changes occur very rapidly, the most conspicuous being the complete
resorption of the large muscular tail. (Fig. 18.4d) The tadpole is transformed into an
immature froglet and metamorphosis is completed. (Fig. 18.4e)

Fig.18.4: Metamorphosis in khe frog. a, Tadpole with rudiment of hindlimb in the fonn of a small bud. b,
Tadpole with fully developed hindlimbs; c, d, stages of metamorphosis; e, metamorphosed froglet
with remains of the mil;J, g, k, three stages of hindlimb development (these fPll between the stage
of a and the stage: of b). (From Witrchl 1956) .-

The length of the larval period in anurans vary. In some species, tadpoles produced in
spring metamorphose during early summer. while in others the tadpole stage may last
for a year or more before the premetamorphic transformation begms. The first sign of
change occurs in the growth or premetamorphic period by the appearance of swellings
on each side at the posterior end of the trunk (Fig. 18.4a). These are the hind limbs
which grow slowly during the prometamorphic period.

The growth period is followed by the prometamorphic period during which the length
.of the hindlimb increases very rapidly relative to the growth of torso (Fig 18.4b. f, g.
h). A few days before the end of the prometamorphic period a number of changes
begin, notably a shift in the position of the anus and a thinning of the operculum on
each side, a process that forms translucent, "skin windows" through which the
forelimbs will subsequently erupt.
Animal Development
This is followed by metamorphic climax, as a result of which the the forelimbs grow
and erupt through the operculum, the horsy beaks of the oral region are lost, the
mouth widens and muscular jaws develop. The eyes are repositioned to a higher level
and develop lids (Fig. 1 8 . 4 &~ d). These changes involve a complete remodelling of
the head skeleton and are adaptive to a predatory mode of life, requiring an aerial
sensory input. The epidermis thickens, hardening the skin and the jelly-like dermis
gets replaced by a tougher more librous tissue. Within the skin, the pigment cells get
arranged in such a manner so as to give the adult pattern of colouration. A muscular
tongue used for capturing prey develops, the hyoid cartilages differentiate, the gills are
resorbed, as lungs for pumping air into the body which become fully functional. The
cells of the larval alimentary tract are almost completely sloughed off and essentially
a new and shorter digestive tract develops. Within a week of metamorphic climax the
tadpole transforms from a tadpole to a froglet. (Fig. 18.4e)
After melamorphosis some anurans adopt a terrestrial mode of life, returning to the
water only lo breed. Others spend a considerable amount of time out of water but
usually remain in a moist environment. Table 18.2 summarizes the changes in anuran
metamorphosis.

Table 18.2 : Summary of Metamorphic Changes in Anurans

1. External Features
TissueIOrgan Changes Function

Head Loss of horny beaks to

mouth.
Widening of mouth. Teeth Adaptation to new diet.
develop in upper jaw.
Development of tympanum, Accommodation to aerial .
Repositioning of eyes. sensory input.
Limbs Complkte growth of fore-. Locomotion on land.
and hind-limbs.
Tail Complete resorption. Loss of swimming as
major mode of locomotion.
Skin Pigmentation changes. Protective colouration.
Hardening. Protection against water
loss on land.
2. Internal Features
OrganISystem Changes Functions

Digestive system Development of long

muscular and bifid .
tongue from the floor of Change from herbivorous lo
buccal cavity. carnivorous diet.
Major shortening of gut.
Repositioning of anus.
Respiratory Resorption of gills. Change from water-to air-
system Development of lungs. based respiratory system.
Development of h yoid
cartilages and muscles
for respiration.
Reproductive Development of gonads. Sexual maturity only in
system adult form.

Sense organs
Eye Development of nictitating To protect eyes in the water.
membrane and eye lid,
Nervous Degeneration of Mauthner Denervatisn of degenerating
system cells. tissue.
 Growth of new neurones Innervation of new Metamorphosis
and nerves. structures.
Lateral line Degeneration of lateral line Not required for terrestrial
organ organs life.

3. Biochemical
changes during
anuran metamorphosis.
-
Exe Change in visual pigment
from porphyropsin to
rhodopsin.
Liver Synthesis of urea cycle Change in excretory product
enzymes. from ammonia to urea.
Synthesis of serum Maintenance of homeostasis.
albumin.
Synthesis of cerulo- . Connected with changed
plasmin. iron utilization?
Eryaropoieeic Change from synthesis Lower affinity oxygen
tissue of larval haemoglobin carrier for air-based
to adult haemoglobin. respiration.
Gut Synthesis of hydrolytic Resorption of tissue.
enzymes.
Appearance of peptic Change to digestion of
activity in foregut. animal tissue.
Skin Melanin synthesis. Protective colouration
Induction of Na-K- Maintenance of
ATPase. electrolyte balance.
Serotonin synthesis.
Changes in collagen Changes in mechanical
deposition and breakdown. properties of skin for
terrestrial life.
Tail Synthesis of hydrolytic Resorption of tissue.
enzymes.

In anurans the metamorphic changes are very striking and almost every organ gets
modified as shown in Table 18.2. The changes that occur during metamorphosis can
be (studiedunder two categories:
I
(i) Morphological changes
(ii) Biochemical changes
i) Morphological changes
Thp changes in the structure of amphibians during metamorphosis aregrouped into
thre'e categories and are described as (a) regressive changes, (b) progressive change (c)
constructive changes or remodelling.
a) Regressive changes-These changes involve the gradual reduction and ultimate
disappearance of all those larval structures or organs which become redundant in
adults. Looking at the table 18.2, the structures which regress can be easily
noted. The ventral suckers, external gills, the long tail with fin folds are
reabsorbed during early functional life. The gill clafts are closed; the
peribranchial cavities, the horny teeth and horny lining of the jaws are lost. The
shape of the mouth changes, the cloaca1 tube shortens and gets reduced. The
lateral line organs of the skin of tadpole disappear and some blood vessels are
reduced.
b) Progressive changes-Some organs and structures become functional during
and after metamorphosis. In the anurans these changes are tabulated in Table
18.2 and involve the development of the fore and the hind limbs, the middle ear
in connection with the first pharyngeal pouch (the pouch situated between the
mandibular and hyoid arches), the tympanic membrane supported by the circular
Animal Devdopment tympanic cartilage. The eye protrudes on the dorsal surface of the head and
develops an upper eyelid. The tongue develops from the floor of the mouth.
c) Remodelling -Some structures and organs which occur and function both
before and after metamorphoses, get transformed or remodelled during the
process in order to meet the requirement of the adult mode of life. These
changes, as given in table 18.2, affect primarily @e skin, intestine and brain.
The skin thickens, and becomes glandular by developing.mulricelhpLr mucous
and serous glands. It also develops an outer keratinized layer as well as
characteristic colour and pattern of pigmentation. The brain gets highly
differentiated. The intestine which was long and coiled in the herbivorous
tadpole shortens and straightens out: Other notable changes which occur are the
change in the blood vascular system in order to supply the lungs, the change in
the portal system, the change in the heart as it becomes three chambered from
being two chambered earlier.
(ii) Biochemical changes
Biochemical changes also accompany morphological changes and are quite striking
in anuran metamorphosis. The changes that occur in anurans are summarized in table
18.2 and are given below briefly.

a) The porphyropsin (a complex between the protein opsin and aldehyde of

Vitamin A2) of the eye during metamorphic climax is replaced almost completely
by rhodopsin (a complex between the protein opsin and aldehyde of vitamin Al)
as the visual pigment. In the eye the larval form of a crystallin in the lens is
replaced by an adult a crystallin that differs markedly in electrophoretic
mobility. An adult form of skin keratin replaces the larval form at
metamorphosis. During metamorphosis large quantities of hyaluronidase are
produced. This enzyme eliminates the hyaluronic acid of the larval skin. This
hyaluronic acid is almost replaced by a mixture of other glycosamino glycans.
Hyaluronidase is not present in appreciable quantities in the adult skin. Other
biochemical changes that occur in the skin as a result of metamorphosis are
alteration in the patterns of collagen synthesis and deposition. This results in a
tougher skin, more appropriate for a land dwelling existence.

b) After metamorphosis, the frog begins to excrete the bulk of its nitrogenous waste
as urea rather than ammonia. Urea like ammonia is very soluble in water but
less toxic. As a result after metamorphosis urea can be formed and retained in
the blood and then be excreted by the kidneys with less water loss than would be
required for elimination of an equivalent quanity of nitrogen in ammonia form.
Production of urea requires the activation of the ornithine-urea cycle in the
liver Fig. 18.5. In this cycle carbon dioxide and nitrogen are excreted in the form
of urea. Metamorphic changes thus involve a reorganization of the metabolic
patterns in the liver for production of the appropriate enzymes of the ornithine-
urea cycle. This reorganization starts very early in metamorphosis even before
there is any apparent change in the body form.

c) At metamorphosis the site of erythropoiesis shifts from the liver to

the bone marrow and spleen. This shift is marked by the production of
haemoglobin with different physiological and electrophoretic properties. In Rana
catesbeiana the shift from production of tadpole haemoglobin (HbF) to adult
haemoglobin (HbA) is essentially complete just before tail is resorbed. Also
during metamorphosis there is considerable synthesis of hydrolytic enzymes
involved in the resorption of larval gut and tail.

d) Degrowth occurs when feeding by the larva is suspended during metamorphosis.

The larva at this stages utilizes the reserve food to produce the energy needed for
completing the various metamorphic processes. As a result there is loss of
weight, with the consequence that the miniature frog produced at the end of the
metamorphosis is smaller and lighter than the mature adult. This reduction of
body mass is termed as 'degrowth'. The reduction in body mass is also partly
due to shrinking of some body parts of the larva. For instance during
metamorphosis the head and trunk become smaller and some parts like the tail
and gills are lost.
1
I
I
e) Autolysis causes the disappearance of larval tail, gills, external gills and fin fold.
The process of autolysis is brought about by active movement of amoeboid
macrophages which phagocytose the debris of disintegrating cells. The lysosomal
enzymes of the phagocytes, in particular the cathepsin, show a high rise in
concentration and utilization. The phagocytosed or autolysed material is
reabsorbed and is used in the construction of new organs of adult.

I f) Enzymes during metamorphosis show a remarkable change. There is a change

both in the types and amount of enzymes produced to help in the completion of
metamorphosis. Some larval enzymes not needed in adult are no longer
synthesized, whereas new enzymes needed in the adult begin to be synthesized.
Metabolism of carbohydrates, lipid and nitrogen undergo changes in the adult.
Carbamoyi-phosphate
NH3 1 Synthase Carbamoyl-phosphate
C02
ATP I Ornithine
~arbamo~ltransferase

Omithine Aspartate

Argininosuccinate
Urea
synthetase
Arginase \ Argininosuccinate
/
Ar .@nine
lyase

Carbamoyl-phosphatesynthas
0 Omithine-carbamoyltransferasc
Argininauccinate synrhetsse
Argininosumnate l y a
..

10 IS 17 20'
Stage of development

Flg.18.5 : Development of urea cycle during anuran metamorphosis. (A) The maJor features of the
urea cycle, by which nitrogenous wastes can be detoxified and excreted. (B) Emergence of
urea cycle enzyme activities correlated with metamorphic changes in the frog Rana
cafeabbno. (After Cohen, 1970)

18.5.2 The process of metamorphosis in urodeles

Metamorphosis in urodele amphibians is considerably less drastic. The larval
salamander (a member of genus Ambystoma) for instance has external gills and a long
tail with dorsal and ventral fins (Fig.18.6). The broad mouth, both of the larva and the
adult remains essentially the same. The forelimbs appear earlier than the hind limbs and
both pairs of appendages grow gradually, independent of any metamorphic stimulus.
Metamorphosis essentially involves loss of external gills and tail fin, development of
lungs, closure of gill slits, appearance of eyelids, formation of maxillary bones,
ossification of skeleton, cornification of skin and differentiation of skin glands.

Fig. 18.6: larva of Ambysloma ,a salamander

Animal Development In urodeles, metamorphosis is more gradual on the whole and may take several
weeks. It involves the following changes:
a) Regressive changes: Tail is retained but fin fold disappears. Branchial apparatus gets
reduced. The external gills get reabsorbed and gill clefts close. The visceral skeleton
becomes reduced.
b) Progressive changes
Head shape changes. The eyes develop lid and bulge more on the dorsal sides
of the head. Skin becomes multilayered and cornified. Its pigmentation changes
and the skin glands also become differentiated. The legs and alimentary canal
hardly undergo any change.
After metamorphosis, depending on the species, urodeles may reduce the time spent in
water or they may become land dwellers returning to the water only to breed. Newts
and Salamanders that become terrestrial undergo a second metamorphosis when they
return to the water for breeding.
Second metamorphosis-Some urodeles like newts and salamanders show two major
changes in form, physiology and life habit between birth (or hatching) and adulthood.
For example, the spotted newt Notophthalmus viridescens, exhibits two prominent
metamorphic changes during its lifecycle. This newt hatches out as an olive green
larva with gills, a keeled tail and a well developed lateral line organ system (a system
of receptors sensitive to local deplacement of water). After a few months of growth it
undergoes metamorphosis, whereby the gills and tail fins are resorbed, the lateral line
system becomes nonfunctional and skin becomes rough and dry and orange in colour.
The visual pigment which was mainly porphyropsin in the larva changes to a mixture
of porphyropsin and rhodopsin. The newt is called an eft and it becomes a woodland
dweller for 2 or 3 years, remaining on land and growing to full size.
The eft phase is terminated when the animal undergoes a second metamorphosis
through the action of hormones prolactin and pituitary gonadotropins. These initiate a
drive towards water which is accompanied by such physiological changes as the
functional reinstatement of the lateral line organ system, the reversion of the skin to a
mucous secreting, wet and shiny organ, the restoration of a finned tail, the maturation
of the gonads and the adoption of porphyropsin as the main visual pigment. In this
manner the newt returns to the water, to spawn and remains there throughout its life
which lasts for several breeding seasons.
Some urodeles, like the permanently-gilledsalamanrlcrsappropriately called
'perinnibranchiate', do not undergo metamorphosis in nature and so grow to sexual
maturity as permanently aquatic animals that retain their larval features like external
gills. Thus they are neotenic and paedogenic. Neoteny is a condition in which the
larval characters are retained for prolonged periods of time. Fig. 18.7 shows some
neotenic urodeles. Paedogenesis is the process by which larval individuals reproduce.
In subsection 18.5.8 you will read in more detail about neoteny and paedogenesis.

Flg.18.7: Some neotenic urodelesThese retain their larval features but are sexually mature
(a) Neclurus (b) Proleus (c) Siren ( d ) Pseudobranchus (e) Amphiuma (f) Crylobranchus

SAQ 3
i) Fill in the blanks with suitable words
a) In anurans, melamorphosis is usually associated with a transition from
.......................to ....................... mode of life.
 b) The axolotl larva of Abystoma achieves, sexual maturity as a larvae. This
phenomenon is known as ........................
c) The salamander Notophthalmus viridescens undergoes a .......................
metamorphosis before becoming adult.
d) The initiation of metamorphosis is due to the ;...................... of the
genomic set by a ......................................
e) The three distinct spges of metamorphic changes in anurans are ................
........................................ and ...................... :.......................
f) The three categories of morphological changes occurring during the
metamorphosis of amphibians are ..............: ...............................................
and .............................................. changes.
ii) Indicate the following changes that occur during metamorphosis in amphibians
either as progressive or regressive or remodelling
a) The development of middle ear in connection with the pharyngeal pouch.
b) The change in the shape of the mouth and the shortening and reduction of
the cloaca1 tube.
c) Disappearance of lateral line organs of skin and reduction of blood vessels. -
d) The differentiation of brain.
e) The changes in the portal system and the changes in the vascular system
to supply blood to the lungs.
f) The shortening and straightening of intestine.
g) The conversion of the heart into a three chambered one.
h) 'The development of fore and hind limbs.
i) Closing of gill clefts and loss of horny teeth.
j) Development of tongue from the floor of the mouth.
iii) List any four physiological changes that occur during the transformation of
tadpole into adult frog.

iv) List any two regressive and two progressive changes that occur during the
metarnorphis of urodeles.

18.5.3 Hormones in metamorphosis of amphibia

In amphibians, the changes that occur during metamorphosis are brought about by
hormonal secretions of the thyroid gland. The first indication of this was obtained
in 1912 when Gudematsch reported that frog tadpoles when fed dried and powdered
sheep thyroid underwent precocious or early metamorphosis. Similar results however
were not observed when tadpoles were fed with preparation of other glands. These
experiments made it possible to postulate that thyroid hormones bring about
metamorphosis. In 1918 B.M.Allen observed that removal of thyroid rudiment from
early frog tadpoles, prevented them from undergoing metamorphosis, and caused them
to become giant tadpoles instead. On the other hand, if they were fed with thyroid or
immersed in water containing soluble extracts from thyroid glands they then
proceeded immediately to metamorphosis. (Fig. 18.8)
Similar experiments in urodele amphibians, in particular in axolotl, Ambystoma
mexicanum, also indicated the importance of the thyroid gland in urodele
metamorphosis.
 Animal Development

Fig. 18.8: Hormones and metamorphosis (a) Normai me(amorphic stage (b) Premature metamorphosis
following exposure of young tadpole to thyroxine. (c) Inhibition of metamorphosis foliowing
removal of thyroid or the pituitary gland.
Later studies by W. Etkin 1968 have also shown the importance and role of different
hormones in metamorphosis. He concluded that development is controlled by a
dynamic balance of plus and minus factors or hormones. Furthermore he found that
the spacing of metamorphic events depends on the concentration of thyroid hormone,
while the sequence of events is inherent in the tissues.
18.5.4 Interaction of amphibian harm-mes in the process of
metamorphosis
pzGGEq
Neuro secretory ails
62 All the diverse changes that occur in anurae metamorphosis are brought about by
the interactions of the hormones secreted by the thyroid gland, hypothalmus of brain
and pituitary.
I *

piiGzGq
5.
THYROID STIMULATING FRm
& HORMONE (TSH)
1
~ S S U E ~ YOU now know the principle organs involved in metamorphosis. Let us see how they
function and coordinate during metamorphosis. Etkin, 1968 has summarized
z amphibian metamorphosis as three main events (Fig.18.9) which are as follows:
(1) The pace of metamorphic events depends on the concentration of thyroid
J hormone. As a result, the level of the thyroid hormone in blood and tissues
THYROID HORMONE gradually increases during the last two third of larval life, up to the event of
+OTHER FACTORS,
?
metamorphic climax, after which it drops suddenly.
(2) At the time of metamorphosis the concentration of thyroid hormone in blood
TADPOLE TISSUES and tissues is increased by TSH-RF secreted by the hypothalmus.
(3) The reactivity of different larval tissues to different concentrations of thyroid
Flg.18.9:Controi of release of hormone is inherent (genetically programmed) in the tissue. This means that
thyroxin during different tissues have different critical threshold.
metamorpbd
The hormonal interactions of metamorphosis are summarized in Fig. 18.10 and are as
follows.

PREMETAMORPH OSlS PROMETAMORPHOSIS METAMORPHIC CLIMAX

(dopamine) (dopamine)

Pituitary P~tuitary Rtuitary

I
I
I
I
I
I
I
I

t
Prolactin

Growth of larva Initiation of -m

metamorphosis
\

Fig. 18.10: The hypothalamus-pituitary-thyroid axis during different stages of annron

metamorphosis. As hypothalamus &veiops, it stimulates the pituitary to in&nct thyrdd
hormone secretion and Inhibit prolactin secreUou T, Trliodothyronlnej To thymine;
TSH, thyroid-stimulating hormone; TSH-RF, t b y d d stimulatirt# hormone reieodng factor.
 The thyroid gland secretes a large complex protein thyroglobulin which has a Metamorpnosis
molecular weight of 675000. The thyroglobulin is formed of several molecules of low
molecular weight compounds. These cornponds are tri-iodo-thyronine (T3) and
thyroxine or tetraiodothyronine (T,). As the figure' 18.11 shows in both these
compounds 2 residues of the amino acid, tyrosine are joined together . Three atoms of
iodine in tri-iodothyronine and four atoms of iodine in thyroxine are attached to the
tryosine molecules. During metamorphosis the iodine containing compounds T3 and
T4 are released from the thyroglobulin.

3,5,3! 5' Tetraiodothyoninc (TI) 3,5.3 Triiodothyronine,(T,)

Thyroxine
Flg.18.11: Formulae d thyroxlne (T,)and Trllodothyronlne (T3

Of the two hormones T3 and T4, T4 is the precursor and T3 is the active hormone and
can cause metamorphic changes at a much lower concentration as compared to T4. In
both anurans and urodeles the thyroid in the larva does produce small amounts of T3
and T4 hormones. However these amounts are overbalanced; in other words their
effect is countermanded by the hormones secreted by the anterior pituitary. The
action of the pituitary has been found to be indirect. It secretes two hormones (1) the
thyroid stimulating hormone (TSH) which is secreted in larval amphibians only at the
onset of the metamorphosis (2) a hormone similar or identical to prolactin whose
activity is antagonistic to thyroxine during larval life. This is because prolactin acts
as a larval growth hormone and inhibits metamorphosis. However at the onset of
metamorphosis the concentrations of T3 and T4 hormones increase to such an extent
that the action of prolactin becomes ineffective, thereby causing the tadpoles to
become frogs and the larval newts to become land dwelling efts.

The release of Tj is under the control of the hypothalamus. It integrates the chemical
information received from the body for determining the time of starting
metamorphosis. During the period of larval growth (premetamorphosis)
hypothalamus is underdeveloped, so it exerts little control over the anterior pituitary
(Fig. 18.10). In the absence of hypothalamic regulation the pituitary secretes high
levels of prolactin and little or no thyroid stimulating hormone (TSH). Thus T3 levels
are low and prolactin levels are high. When the hypothalamus develops, its
production of thyroid stimulating hormone releasing factor (TSH-RF) increases
causing an increase in the level of TSH. This causes the thyroid to release more T3
and T4. The concentration of T3 gradually increases until the first changes of
metamorphosis (prometamorphosis) appear. During this period the hindlimbs begin to
enlarge. The increase in T3 and T4 titres as well as hypothalamic secretion, a
substance (most possibly dopamine), inhibit the pituitary synthesis of prolactin. The
ratio of T3 to prolactin thus changes and Tj concentration increases enormously. This
leads to metamorphic climax in which drastic morphological and biochemical changes
.
associated with metamorphosis occur along with partial regression of thyroid gland.
The hypothalamus now diminishes its output of factors, affecting the pituitary
hormones, and a hormonal balance appropriate to the life of the growing froglet ensures.

As you know, in certain urodeles the first metamorphosis results in the development of
land dwelling eft, which after some time undergo a second metamorphosis in order to
be able to return to the water to breed. The second metamorphosis has been shown to
be under the influences of prolactin, presumably resulting from a shift in balance
between pituitary factors prolactin and TSH rather than an activation of one of them.
In other words, the first metamorphosis is induced by a shift in favour of TSH, the
second metamorphosis by a return to predominance of prolactin.

18.5.5 Tissue reactivity

The mechanism of metamorphosis in anurans indicates that a single agent, namely
I thyroxine, evot?.: rr.;rltiple responses in different tissues. In other words, the various
Anlmd Development organs of the body respond differently to a single hormone agent. In addition, the
response of the tissues, though diversified, are specific and can be destructive or
constructive depending on the target tissue. Thus the same stimulus will cause certain
tissues to degenerate and others to grow and differentiate.

Such diverse reaction of tissues is believed to be due to two reasons:

(1) Competence of larval tissues or multiple response. The reactivity of the tissue
of the target organ to the thyroid hormone.
(2) The threshold value of different larval tissues for thyroid hormone
concentration.

(1) Competence of larval tissue or multiple response. The responsiveness of

different larval tissues to thyroxine is markedly different. The tissues of the tail
becomes necrotic and undergo degeneration due to the action of the thyroid hormone,
while the tissues of the limb increase and undergo differentiation. Furthermore
thyroxine causes rapid aging and destruction of R.B.C's of the larva undergoing
metamorphosis. In addition it stimulates the development of cells that synthesize the
haemoglobin of adult frog. The hormone T3 reduces biosynthesis of nucleic acids in
the tail but increases their synthesis in the liver. Furthermore, muscles of the tail
undergo degeneration while those of the trunk remain unaffected.

Such examples clearly indicate that the response and reactivity of larval tissue to the
same hormone or agent differs greatly. In addition it can also be demonstrated
experimentally that the reactivity and response of the target tissues are intrinsic,
specific and independent.

This can be demonstrated by transplanting tail tips of the tadpole to the trunk of
another tadpole undergoing metamorphosis, or by placing the eye of the tadpole in
the tail of a metamorphosing larva (Schwind. 1933, Geigy. 1941). The extra tail
transplanted into the tadpole host trunk is not protected from degeneration and
undergoes necrosis along with the host tail and gets absorbed. The eye retains its
integrity despite the fact that it lies surrounded within the degenerating tail. (Fig. 18.12)

Eye cup transplanted It)),

to regressing tail
Flg.18.12: Organ speclflclty durlng frog metamorphosis. (a) Tail tips regress simultaneously with the hosts
tall even when transplanted to the trunk, whereas eye cups (b) remaln Intact even when
transplanted Into the regrerslng tall.

The degeneration of tissues during metamorphosis thus represents genetically

determined cell death. In humans such programmed degeneration occurs in the
tissues between our fingers and toes. The degeneration of the human tail during the
week four of development resembles the regression of tadpole tail.
Such observations force us to question as to how does the thyroid hormones bring about
diverse metamorphic changes? Do the hormones act directly on all the target tissues or are
their actions mediated by some other hormone/honnones? Finally, how is the changing
thyroxine tiues controlled? Such questions have been answered by experiments which have
clearly shown that the hormone acts directly on the target tissue. Weber (1967) demonstrated
this fact when he cultured excised tadpole tails in the presence of T4. (Fig.18.13) While tails
treated with thyroxine showed regression similar to that occuning in metamorphosis, the
controls (not treated wilh thyroxine) exhibited no regression and remained healthy.
 Metamorphosis

(a) Control Tips (b) Test Tips

6 Days

8 Days

10 Days

12 Days

Flg.18.U: Regression of Isolated tail ends under the Influence of thyroxine (a) control tail t l p cultured In
Holtfelter7ssalt solution for 6,8,10 and 12 days (b) Treat4 tall tips at the same ages os controls
but having thyroxine added to their solution. .
The regression of the tail is believed to occur in four stages. First, protein synthesis
decreases in the treated muscle cells of the tail. Then, there is an increase in the
lysosomal enzymes. Concentrations of Cathepsin D (a protease), RNase, DNase
collagenase, phosphotase, and glycosidases all rise in the epidermis, notochord and
nerve cord cells. Cell death is probably caused by the release of these enzymes into
the cytoplasm. The epidermis helps to digest the muscle tissue, probably by releasing
these digestive enzymes. After this death, macrophages collect in the tail region,
digesting the debris with their own proteolytic enzymes. The result is that the tail
becomes a large sac of proteoltic enzymes (Fig. 18.14). If the epidermis is removed
from the tail tips, the tips will not regress when cultured in ~hyroxine.
2. Threshold value of different larval tissues for the thyroid hormone concentration
One of the major problems of metamorphosis is the coordinalion of developmental
events. The tail should not degenerate till some other means of locomotion, namely the
limbs develop, and the gills should not regress till the animal can use its newly
developed lung muscles.
Kollros (1961) demonstrated the possibility of this coordination which shows that
different larval tissues have different threshold value for thyroxine. In other words the
different larval tissues are sensitive to different concentrations of thyroid hormone.
This model is called the threshold concept.
It has been observed that structures in the tadpole which change early during
metamorphosis are more sensitive to and have low threshold value to thyroxine, than
those which undergo transformation at a later stage. As metamorphosis
commences the thyroid hormone level gradually builds up and different events occur
at different concentrations of the hormone. Experimental studies have revealed that
tadpole pans which have a low threshold respond earlier during metamorphoses than
those parts having a high threshold response. In other words, the threshold value
reflects the order in which metamorphic changes occur in normal development. In
urodeles the bulging of eyes react to the weakest dose of thyroid hormone (minimum
threshold) and so is the first event in metamorphosis. This is followed by reduction of
fin fold and the shortening and disappearance of external gills. After which occurs the
closure of gill clefts and transformation of skin.
Anlmnl Development

Relative length of tail (%)

Flg.18.14: Regression of tadpole tail, in metamorphosing of Xcnopus hcvis LF accomplished by lysosom

PI digestion of cells. As metamorphosis proceeds, the enzyme concentration Increases (the absolute quantity
of enzyme remains constant). Eventually the stub contains almost nothing but lysosomal enzymes
and it falls off.

The administration of high amounts of thyroxine at early stages causes precocious but
distorted sequence of metamorphic changes resulting in death. Tissues. responsive to
low concentrations will not respond, while those responsive to high amount do so
precociously. For example; in the frog tadpoles as hormone concentration increases
tissue responses become progressively more rapid till maximum rates of change are
attained. When the dosage is heavy all metamorphic events start together or crowd
together and the normal sequence of events is disturbed. The destructive processes
being capable of proceeding faster than the constructive processes result in the forelimb
erupting before becoming differentiated; the tail becomes reduced before the legs are
sufficiently developed to take over locomotion-resulting in an abnormal animal which
dies. There is a progressively increasing threshold of the hormones at different stages
of development of a particular organ and during the entire metamorphic process.
18.5.6 Induction in metamorphosis
Some of the morphegenetic changes during metamorphosis are found fo be quite
independent of hormone action. For example, usually during metamorphosis, the skin
of the tail undergoes &generation under hormonal effect. It is observed that when the
skin alone is removed from tail and transplanted onto the tail of another
metamorphosing larva, it does not regress despite the presence of the hormone. But if
the skin is grafted to the trunk along with its underlying muscle in a developing tadpole,
then it regresses. Thus the hormone affects only the muscle directly, which induces
regression or progression of the skin depending upon the induction it receives from its
underlying muscle of tail or trunk respectively. Similarly tympanum is another example
of the induction process and is independent of direct hormone, action as its formation is
induced by the tympanic cartilage.
18.5.7 Molecular response to thyroid hormones during
metamorphos~s
Thyroid hormones can cause existing tissues to break down or can remold the tissues
tq their adult function. The liver cells of the tadpole for example are not destroyed
and replaced during metamorphosis. Instead their structure is remodelled. This
change is accompanied by dramatic increase in ribosomal and messenger RNA
synthesis. The rate of protein synthesis also increases nearly 100 folds within four
hours of thyroid hormone stimulation. Many of the new mRNAs are those which code
for new functions of the liver.
The major increase in protein synthesis appears to come from the transcription of new
mRNAs. As Fig.18.15 shows, carbamoylphosphate synthase (a urea cycle enzyme) is
synthesised after the burst of RNA synthesis. Experiments have demonstrated three
types of molecular response to thyroid honnone. One set of genes increases its low
level of transcription in response to either natural or thyroxine induced
metamorphosis. Another set of genes decreases its rate of transcription. while a hird
set of genes remains unaffected by thyroid hormones. Mori and Coworkers (1979)
have demonstrated that much of the increase in carbamoylphosphate synthase can be
attributed to increased transcription from the gene. Thus metamorphosis appears to
some extent to be controlled at the transcriptional level. Other evidences also indicate
the ability of thyroid glands to regulate gene activity at the level of transcription.
This does not mean hat transcription is the only level of gene regulation, operative
during metamorphosis, but it is obviously an important one.

Nuclear RNA Carbamoyl-

phosphate
,

Cytophasmic RNA

o &albumin

0 12.5
Time after treatment with triiodothyronine (days)

Fig.lll.15: MdcarlPr syntkk~La Rtma ccltrrbiane liver cells after treatment of tadpdes with hllodoth~ronfne.
Fh-slan inacPee In the nudear and tbco In qmplasmk RNA are obeewd before on LnerwesLa Uver-
q d I c pmkh,.espedslly those ol the urea cycle (After Graham and W a 1976)

18.5.8 Neoteny
Urodele metamorphoses has shed light on certain aspect of gene expession and tissue
response during metamorphosis. A number of urodeles undergo delayed, partial or no
metamorphosis. Trituris vulgaris has a tendency to retain a few or more larval
characters in the adults. This is called neoteny. Many salamanders are able to retain
their larval form throughout their lives, becdming sexually mature without undergoing
metamorphosis. The degree to which metamorphosis occurs varies from species to
species. The Mexican axolotl Ambystoma mexicanurn does not undergo
metamorphosis in nature. remaining a sexually mature animal with larval characters
such as external gills and a long tail with dorsal and ventral fins. The animal remains
neotonic as it does not release active TSH to stimulate its thyroid glands. However
when investigatars gave A mexicanwn either thyroid hormone or.TSH hey found that it
metamorphosed into an adult not usually seen (Huxley 1920). (Fig. 18.16)

Other species such as Ambystoma tigrinurn metamorphose only if given cues from the
environment. Otherwise they become neotenic, successfully mating as larvae. Some
perinnibranchiates that remain permanenlly gilled do not even respond to thyroxine or
iodine tteaunent in the laboratory and so remain permanently neotenic. The tissues of
these animals consist remarkably of comparatively enormous cells. as though the larva
had grown to adult size by the process of cell enlargement instead of cell proliferation.
The neotonic species of Necturllr and Siren also remained unchanged despite thyroid
honnone treatment, indicating that h e larval tissues in these cases have lost their
capacity to respond to thyroid hormone. Thus their neoteny is permanent (Frieden
1981). The genetic lesions responsible for neoteny in several species are shown in
Fig.18,17.
De Beer, 1940 and Gould, 1977 have speculated that neoteny is a major factor in
the evoludon of more complex taxa. By retarding development of somatic tissues,
natural selection is given a flexible substrate to act upon. According lo Gould, neoteny
would provide an ecaspe from specialization. Animals can slough off their highly
specialized adult forms, return to the lability of youth, and prepare themselves for new
evolutionary directions.
 Anlmal Development

Thymtmpin releasing
hormone (TSH-RF)
1

+-
Flg.18.16: Metamorphosis Induced In axolotl (a) Normal condltlon of axolotl (b) Anlmal treated with
thyroxlne to Induce metamorphosis.
Thymid
stimulating hormone
SAQ 4
EuFa neotenes 1) Indicate whether the following statements are true or false. .
Thymid a) Tadpoles when fed with dried and powdered thyroid gland from any other
animal will undergo an early and precocious metamorphosis. TrueFalse
I
mymune b) The sequence of metamorphic events depend on the concentration of
Triiodothymnine

1 . thyroid hormone, whereas the spacing of the events is inherent in the

tissues. TrueFalse
c) TSH-RF secreted by the pituitary gland activates the thyroid gland to
Target tissues
swrete thyroid hormone. TrueFalse
Capable of undergoing
metamorphosis
1 d) Tetraiodothyronine (T4) is a less potent hormone and is the precursor of
triiodothyronine (T3) TrueFalse
~ig.18.17:stages along the e) Prolactin has an antagonistic action to thyroxine, and so promotes larval
hypothalamus-pltultary- growth and inhibits metamorphosis. TrueFaise
thyrold axls of
salamanders at which f) Thyroxine, the single hormonal agent evokes multiple responses in different
varlous specles have tissues and its action whether constructive or degenerative, depends on the
blocked metamorphqls.
Euryeea, Necrurus, and target tissues on which it acts. TrueFalse
Siren appear to have a
receptor defect In the g) The reactivity and responses of target tissues of the tadpoles to the
responsive tksues. thyroxine hormone are extrinsic and non-specific. TrueFalse
~ u b c e wlll
a
metamorphose when h) Genetically programmed cell death is responsible for several of the
treated with high degenerative changes that take place during metamorphosis. TrueFalse
eoneentratlons of
thvroxlne. Necrurus and i) According to the threshold concept, different tissues of the tadpole larva
~in'ndonot r e s ~ o n dto anv are sensitive to dilferent concentrations of the thyroxine. TruePalse
dose. (After Fieden, -
1981). j) In tadpoles the regression and the progression of all tissues is dependent
. totally on the action of thyroxine and the induction by underlying tissues
do not have any role in the metamorphic process. TrueFalse
ii) Match-the statement on the right side (A) with the species on the left (B).

a) No metamorphosis in nature. i) Ambystoma tigrinurn

retention of all larval ii) Necturus
characters but sexually iii) Ambystoma mexicanurn
mature. iv) Ambystoma mexicanurn
b) Metamorphosis occurs only if Metamorphosis
environmental cues are provided.
Otherwise neotenous larvae successfully
reproduce

c) Administration of thyroxine or TSH

results in the metamorphosis of the
otherwise neotenous lama into an adult

d) Administration of thyroid hormone has

no effect on the larva as the larval
tissues have lost their capacity to
respond to thyroid hormone. These
forms show permanent neoteny.

Moult
18.6 DEVELOPMENT, GROWTH AND
METAMORPHOSIS IN INSECTS
Instar
18.6.1 General process of post-hatching growth in insects
When a young insect hatches out from an egg it is covered by a firm, inflexible,
sclerotized cuticle, which due to its rigid structure cannot grow along with the increase
in length of the hatched larva. Thus during development, for the hatched larva to
attain its adult size and form, the old rigid cuticle at every stage of growth has to be
replaced by a newer, larger one by the process of moulting. Accordingly development
is marked off by a series of moults.
The interval between the two moults is known as stadium and the form that an insect
assumes as a result of the moults is called instar. All insects undergo several moults
after hatching from the egg and before emerging as adults which are called imago.
The number of moults is 4 or 5 in an insect species and is usually fixed or
predetermined, but it is not absolutely constant. Furthermore, the form of the insect
changes with each moult in a precise pattern characteristic of the species. Moulting is
not just a mechanical process; instead at each moult, the moulted insect undergoes
changes both in its cuticular covering and internal organization. So you can see that
development in insects is usually by the process of metamorphosis. The metamorphic-
changes may be siight and gradual or radical as occurring during the pupal stage. Flg.18.18: Development of an
During this stage the lama remains quiescent without feeding or movement, while Insect (the silk moth
many larval characters disintegrate and adult structures are formed. Pupal stage Cecropia) wlth complete
metamorphosls.
follows the last larval molt and from the pupa the imago emerges. Fig. 18.18 shows
metamorphosis in the silk moth, Cecropia.

18.6.2 Patterns of metamorphosis

The pattern of metamorphosis is not the same in all insects and so metamorphosis can
be broadly classified into 3 types:
(1) Slight or no metamorphosis as observed in Apterygota-ametabolous or direct
development.
(2) Incomplete metamorphosis as observed in Exopterygola-Hemimetabolous
development.
(3) Complete metamorphosis as observed in Endopterygola-Holometabolous
development.
Animal Development In Table 18.3 you.can see how insects are classified on the basis of types of
metamorphosis in their life history.

Table 18.3: Classification of insects based on types of metamorphosis

Class Insccls
Subclass Pterygota / Apterygota
(Winged, metamorphosis) (wingless, metamorphosis
slight or
absent; most primitive
insects)
e.g. silver fish, spring tails.

Exopterygota / prcscnt \ Endopterygota

Hemimetabolous metamorphosis Holometabolous metamorphosis
a Wings develop externally a Wings develop internally
a Miniature stages-nymphs, naiads; a Immature stageslarvae
resemble adu1ts.e.g. Cock- differ in structure and
roachs, grasshoppers. function from adults. e.g. butterflies,
moths, ants, bees, beetles.

(1) Slight or no metamorphosis

In primitive wingless insects (Apterygota) like spring tails, silver fish etc. and in
secondarily apterous insects the young ones that hatch from the egg are essentially
similar to imago, differing from it only in size, immature reproductive organs and
external genitalia. It moults several times leading to a gradual increase in size
accompanied by the appearance of the external genitalia and maturation of sex-organs.
In apterygotes, growth and moulting are seen to continue even after reproductive
maturity. Such a type of development or life cycle is called ametabolous. Fig.18.19
shows such a metamorphosis in silver fish (Lepisma)
Egg -> Young goes through several stadia and moults -> Imago (Adult)

Fig.18.19: Ametabolous type of development in Lepisma

(2) Incomplete metamorphosis

In the Exopterygote insects rudiments of wings are present only as buds at hatching
and the body form is disproportionate to that of the adult. As the moults occur the
form and size of the larva progressively approach that of the adult. The wings become
fully developed and sexual maturity is achieved at the last moult. The degree of
metamorphosis in the hemimetabolous forms are of two distinct types, namely gradual
metamorphosis and extensive metamorphosis.
(a) Gradual metamorphosis - In Exopterygote insects like grasshoppers, crickets,
cockroaches etc., the nymph is similar in both structure as well as in habit to the
adult, but lacks wings, gonads and external genitalia. It undergoes several moults as
it grows and develops genitalia, gonads and wing pads in the later larval period. The
wing pads get transformed into functional wings at the last moult, after which no
more moults occur. This type of metamorphosis has no quiescent stage and there is
 no loss or remodelling of larval parts. Fig. 18.20. Metamorphosis

Egg -> Nymph ->several instars and moults---> Imago (Adult)

FLg.1820'shows padual metamorphosis In grass hopper. There are five nymphal stag- k l o n the adult
condltlon l a reached.

(b) Extensive remodelling -

In insects like damsel flies, mayflies, dragonflies
etc.. the nymph differs considerably in both external structure as well as in habits
from the adult. The nymphs are aquatic and herbivorous and possess some organs for
locomotion in water, tracheal gills for respiration and mandibulate mouth parts for
I cutting vegetation. Such aquatic larval forms are called naiads. They undergo
extensive remodelling to assume the adult form. They shed gills and modify their
mouth parts and develop wings. Moulting may lead to one or more quiescent or
! semiquiescent larval stages.
I
b This type of metamorphosis is called incomplete extensive metamorphosis Fig.18.21.
\
t
Egg -> Naiad -> several moults -> Imago (Adult)

r-
Matlng on wiag

7 .
in midsummer

Imago f IIW away Female ovipositor makes

SINS UI leaves of aquatic

I plants and lays eggs there

t
I
Wings d y cuucle hardens

t
Final ecdysls

Immobile for
Falling temprature in
autumn kills adults
-'
I
' I weeks
th-
Muatic nymph (naiad)
without mngs emerges
gills m rectal region
several how+
\
When wings a p p x i m a t e l ~
used for gaswus

1 em long nymph climbs

up on t o - suitable plant First year
for suppn out of water feeds and grows . many moults dunng
which wngs gradually develop
nymph
NB nYmphs resemble the adult (image) in body form out p m w s adapmtlons
whxh t lt them to live m water,a habitat quite d~fferrntt o tne temstnal environment
ot the imago-incomplete mctamorphos~s

adult

Flg.1821: Shows Incomplete extensive melamorphosls In dragon fly.

Pupalion is a metamorphic moult
(3) Complete Metamorphosis and may ~nvolvethe formation of
spec~alpupal cuticle as in
In all Endopterygote insects, in which wings and other structures develop internally, butledlies and moths which spin
(in invaginated imaginal epidermal pockets) like beetles, wasps, bees, butterflies, a cocoon and pupate within it. In
moths etc., the larva which hatches out of egg is very different from the imago, in others the Pupa remain in h e
old larval skin which forms the
habit, appearance and structure. The larva has a worm-like body, biting and chewing puparium.
mouth parts, simple eyes and weakly developed walking legs. It has quite a different
29
Anlmd Devdopment habit. For example the mosquito larva lives in water and feeds on protozoa and algae,
while the adult either sucks blood or fruit and flower juices. A more common example
is the butterfly larva which crawls on the ground and feed on leaves and then gets
transformed into an aerial organism feeding on nectar from flowers.
The larvae of these types of insects are either swimmers or crawlers, and are voracious
eaters. They grow in size and moult several times till they attain a quiescent, non-
feeding stage called pupa. The pupa is enclosed in a pupal case or the puparium
secreted by the labial glands of the larva. This pupa does not move or feed and its
energy must come from the nutrients it ingested while a larva. Externally it appears
as an inactive structure. However internally it undergoes basically two types of
changes at a rapid pace and moults only after the complex series of internal changes
have taken place. These changes involve wholesale destruction of most of the larval
tissues (histolysis) and the formation of an entirely new adult body whose organs and
systems are developed (histogenesis) from nests of organ specific cells, called the
imaginal discs. Histolysis results in systematic destruction of the old body of the
larva so that all the organs except for the central nervous system are broken down by
special amoebocytic cells called phagocytes. The tissue fluid, which arises due to the
destruction is used as raw material in +e formation and histogenesis of the adult
organs. After these changes are completed as a result of histolysis and histogenesis
the pupa undergoes the pupal moult and the imago (adult) emerges fully ready to lead
a short or long independent existence and to reproduce. This type of metamorphosis is
called complete metamorphosis as shown below and given in Fig.18.22.
Egg -> Larva -> several instars and moults -> Pupa -> pupal moult->
Adult.
th
5 ~nstar

,. .....

....,..
. ...
,.......
. ..

Moult

Flg.18.22:shows complete metamorphosb In silk worm moth.

Imaginal Discs: In the holometabolous larva, there are two cell populations: (1) the
larval cells which are used for the larval structures and (2) the imaginal disc and the
histoblasts which are present in clusters, awaiting the signal to differentiate. Imaginal
discs do not occur in the nymphs and larvae of hemimetabolous insects.
The imaginal discs or buds are actually rudiments of future organs of the adult, such
as mouth parts, wings, antennae, walking legs, and internal organs etc. Fig.18.23
shows the imaginal discs in the larva and pupa of a fly (Musca).
These discs develop directly from the eggs and remain nonfunctional throughout the
larval stages. During the pupal stage they grow in size and differentiate to form adult
structures which remain collapsed and folded. When the reorganization is completed
the pupa moults to set free the adult or imago. Fig 18.19 and 18.22 show the process
of complete metamorphosis. Once the adult or imago blood is pumped into these
collapsed structure it causes them to unfold and inflate. Furthermore chitin is
deposited on then to harden them.
 (d)

Flg.18.23: Imaginal dlses in larvae and pupae of Musca, (fly) a: me imaginal discs of thelarval digestive tract
b: dose-up of the larval proventriculus showing imaginal cells of the stomodaeum. c, d, c:
Imaginal dl- of the legs, and head parts seen from the left side in c for the larva and in d and
c for the pupal stages. (AT) antennal imaginal discs; (B) brain; (E) esophagus; (IM) Imaginal cells
ofmld gut epithelium; IMU, Imaginal cells of mid-gut muscles; (IPA) posterior abdominal imaginal
disc; (PIR) imaginal disc of proctodaeum, L1,L2,L3,discs for the first, second, and third legs;
(OD) optic dlsc; (PB) proboscis; (PD) proctodneum; (PR) proventrlculus; (S) sucking stomach;
SG,salivary ghnd; I, 11,111, flrst, sceond, and third th,oracic segments

The mode of development of imaginal disc varies from species to species and also
from organ to organ. Imaginal discs are sometimes formed during late embryonic
development and their cells are separate from the prospective larval cells, as for
example in Drosophila and other Diptera. In some insects the imaginal discs are
derived from larval cells during the later phase of larval growth.

18.6.3 Factors controlling metamorphosis in insects

For a larval moult to be successful all parts of the body must take part in the process
and carry it out at the same time. This indicates that a common factor acts on all parts
of the body. The existence of a common factor or cause is even more apparent in
metamorphosis, in which the involvement of both external and internal organs may be
more radical and faneaching. This common factor may be external or internal.
External factors
In some cases an external'factor may be responsible for initiating moulting, as for
instance in the blood sucking Rhodnius the intake of food (blood meal) is such a
factor. Another example in which external factor is essential to initiate a moult is the
case of the pupa moth Platysamia cecropia. After pupation the insect falls into a
quiescent state with a reduced rate of metabolism - diapause - which continues
throughout winter. It is essential that during this time the pupa be exposed to cold,
otherwise the diapause is prolonged indefinitely. The diapause may be broken
precociously if the pupa is exposed to cold (3' to 5' C) for at least two weeks. The
temporary cooling activates the vital processes in the pupa and on return to a warmer
environment development is completed, the pupa moults and the imago emerges.
In other insects factors such as humidity, population density etc., appear to initiate
metamorphosis. However in the majority of insects no external cause of any moult
has been detected and moults follow one another at intervals which appear to be
determined entirely by the internal processes in the animal.
Animal Development Internal factors
Similar to the amphibian metamorphosis, moulting and metamorphosis in insects has
been found to be initiated internally by hormones.
Hormonal control involves at least three organs of endocrine secretions . They are (1)
brain (protocerebrum) (2) corpora allata (3) prothoracic gland. Fig 18.24 shows the
endocrine glands associated with the brain in moths namely protecerebrum and

nsc
t b>
FLg.18.24: The endocrine glands assoclated with the brain in meal moths. a) Dorsal vleW of the braln In a
meals moth pupa, b) Transverse section of the protecerebrum in a caterpillar of the meal moth. Ao, Aorta;
br, brain; ca, corpora allata; cc, corpora cardiaca; lg., loregut; nsc, neurosecretory cells.

18.6.4 Organs and hormones involved in insect metamorphosis

(1) Brain neurosectory cells and their hormones

Kopec was the first to suggest the role of hormones in controlling metamorphosis. On
the basis of his experiments on the larval gypsy moth (Portheria dispar) he observed
that at a particular period the brain releases a substance into the blood which is
essential for pupation and hence for metamorphosis. His findings on the role of brain
hormones in metamorphosis of insects was supported by subsequent workers who
found similar mechanisms in different insect groups. Large secretory nerve cells in the
brain of the insects called neurosecretory nerves were also identified as being
responsible for affecting pupation.
The secretion of these cells is called activation hormone (AH) or brain hormone (BH).
The activation hormone is comprised of proteins or Lippoproteins. The AH or BH
after synthesis pass along the axons of these cells and end blindlv intn a nair nf
storage and release organs called the corpora cardiaca (CC), located in the
posterior brain. The CC releases the active hormone material called
prothoracotropic hormone (PTTH) which is a small polypeptide. This
prothoracotropic hormone acts upon the prothoracic gland, causing it to secrete the
moulting hormone called 'ecdysone'.
Corpus allatum (Singular) and Juvenile hormone.-The corpora allata are rounded
glands attached to the posterior side of each corpus cardiacum forming a compact
body just behind the brain. In some (Hemiptera, higher Diptera) they are fused to
form a single median structure. Experiments by Wiggleworth have shown that the CA
secretes a hormone which determines the character of each larval instar by limiting the
degree of diffe entiation towards the development of the adult. He named this
f
hormone as ju enile hormone (JH). This is also known as neotenin (youth
substance), (Wigglesworth 1954) and gonadotrophic hormone (Englemann 1957). The
juvenile hormone is similar in structure to terpenes. A large number of compounds
with juvenile hormone activity have been isolated and many have been synthesized.
Some synthetic ones appear more potent than the naturally occurring ones. The
juvenile hormones ensures the occurrence of larval moults and inhibits metamorphosis.
Its presence or absence determines whether the larva will moult into a larval/pupal
stage or into adult stage. The major natural hormone isolated from the adult male
Hyalophora cecropia moth has the following structure (Roller at a1 1967) Fig.18.25.
 Metamorphosis
CH,

cH, GH, CH,

I
o
*F \ I
CH,
C CH,
t-~CH,
CH?' I\ / CH, CH
0-CH

.
Fig.18.25: Juvenile hormone isolated from Cecropia moth

The role of the corpus allatum and the juvenile hormones in metamorphosis has been
proved by several experiments (Fig. 18.26). It has been observed that if an extra
corpora allata from a second stage larva is transplanted to the fourth instar last larval
instar then the moult does not produce a pupa or adult, instead an oversized larva
develops. Removal of corpora allata early in larval life results in premature
metamorphosis .

.18.26: Experiments on effect of moultlng hormone on pupa size a, b, pupae of moth, resulting from
the removal of the corpus allatum (source of the moulting hormones from third-and fourth-
lnstar larva) c, A normal pupa. d, A glant pupa produced by implantlng a n extra corpus
allatum from a young larva into one that had already reached the stage at which it would
normally pupate.

Prothoracic gland (PTC)or moulting gland and ecdysone or moulting hormone

The third endocrine gland-prothoracic gland- is an irregular branching mass of
glandular cells located in the prothorax (segment of the thorax that bears the anterior
most of its three pairs of legs), in close association with the tracheal lubes and
secretes the hormone ecdysone which has been isolated and crystallized in pure form
from Calliphora and from pupa of a silkworm. It is a unique water soluble steroid
and is related closely to cholesterol (Fig.18.27). Recent studies however show that all
a ecdy~oneis converted into P ecdysone (ecdysterone) after liberation into the
haemolymph and that ecdysterone is the active moulting hormone.

0
Ecdvstcronc

Fig.18.27: Commonly occurring moulting hormones (a) (ecdysone) (b) ecdysiero?e, in insects undergoing
metamorphosis
h i d Development Ecdysterone exerts its effect directly upon those cells concerned in growth and
moulting; it activates them and stimulates them to renew protein synthesis. It is
believed that it acts directly upon those loci in the chromosomes which are concerned
with growth. It induces characteristic puffing in the giant polytene chromosomes of
Diptera. These puffs are considered to be the sites for the formation of the messenger
R N A needed for protein synthesis.
18.6.5 Interaction of insect hormones i n the process of
metamorphosis
We have discussed the organs and the hormones generally involved in metamorphosis
of insects. This is because despite the fact that there may be around 1 million
different insect species, there is a striking similarity in the endocrine function of
different hemimetabolous and holometabolous insects. Fig.18.28 shows a schematic
outline of the interaction of hormones in the metamorphosis of insects.
On the basis of figure 18.28 let us study how the actions of the various hormones
bring about metamorphosis in insects.
The moulting process, the start of metamorphosis, is initiated in the brain. The
s t i d u s (as you already know) may be neural, hormonal or environmental, and causes
the neurosecretory c d s of the brain to release the activation hormone which after
synthesis @anges into the active hormone called prothoracicouopic hormone (P'ITH).
The PTI'H stimulates the prothoracic gland to produce ecdysone. Ecdysone after
being converted into its active form, the ecdysterone, stimulates growth and causes the
epidermis to secrete a new cuticle, initiating the moulting process. The ecdysterone
further stimulates the epidermal cells to synthesize enzymes which digest and recycle
the components of the cuticle. As long as the juvenile hormone is present, the
ecdysone-stimdlated moult results in a new larval instar. In the last larval instar stage,
the synthesis of juvenile hormone is reduced, causing its levels to drop below a
critical threshold value. This again

.'
Regulation

Juvenile
hormone (JH)
q - Brain
Neurosecnetory cells

/ Corpus
Corpuscardiacum
allaturn

ProthoracicotK,pic
hormone (m)
. .
Binding protein
gland

Larval structures Pupal structures Adult structures

t I
1 cu tide
I

&Pa ~ d ~ l t

Fig.1828: SchemaUc diagram Illustrating thc control olmoulUng and metamorpboslsla the tobacco bornworm
moth. (After Gilbert and Goodman, 1981).
triggers the release of PTTH from the brain. The PTTH in turn, stimulates the Melmorphosis
prothoracic gland to secrete an unusally large quantity of ecdysone. The resulting
ecdysterone, in the relative scarcity of JH, causes the instar to pupate. In other words
the occurrence of the subsequent moult in the larva in the relative scarcity of JH and 5
abundance of ecdysone, shifts the organism from larva to pupa. During the period of
pupation the corpora allata do not release any juvenile hormone and the ecdysterone
stimulates the pupa to metamorphoses into the adult insect.
18.6.6 Effect of metaqorphic hormones on gene expression in
moulting and metamorphosing insects
The first report of a specific hormone being able to influence specific gene loci,
control their transcription and finally influence cell differentiation was reported in the
1960s by Clever and Karlson on the basis of their studies on Dipteran larva
Chironomus. These findings were based on experiments in which Clever injected
minute quantities of ecdysone into the larva and observed within fifteen minutes,
hormone induced puffings (swellings) at specific regions of a particular chromosome.

Clever also studied patterns of puffing in polylene chmmosones at various larval

developmental stages. He further established that puffing of specific chromosomal
bands takes place at specific times in normal course of development and follows a
fixed sequence whereby certain bands puff at one stage to synthesize RNA and regress
at a later stage. For example he injected ecdysone into an intermoult larva in which Flg. 18.29: Pumng In l a r c t
puffing of a band 1-19A was known to be particularly active, and observed that in 10- bands (numbered) canprominent
be seen In
15 minutes, puff 1-19,A regressed, band 1-18C started to puff, followed by puffing of ch,om,some~
band IV-2B. This puffing sequence was exactly similar to the sequence that occurs
just before moulting, a time at which the insect spontaneously releases ecdysone into
the haemolyph Fig.18.30 shob similar findings in Drosophifa as observed by Becker,
1959.

Flg.1830: Puffs appearing and dlssappearing during the third iarvai instar and the prepupal stage at the
base of chromosome arm 111 L of Drosophib mehnogaster salivary glands. Numbers lndlcate
hours before or aner pupanum formation. After H. Bccker. Chromosome. 10-654 (1959).

On the basis of such experiments it appears reasonable to assume that the hormone
ecdysone somehow turns on the transcription of one or several genes in the larva
(which is apparent due to the occurrence of puffs)
The action of the ecdysone, as to whether it acts as such on the gene or changes the
internal nuclear environment is still not well understood and so several theories have
been advanced. Kroeger (1968) found that ecdysone causes the cell to accumulate
potassium ions (K+), since the primary sets of puffs can also be induced without
ecdysone by increasing the intracellular or inuanuclear concentrations of K+. Thus the
stimulation of K+ uptake by cell and by nucleus may be the primary effect of
ecdysone.
..
Goodenough and Levene (1974) have suggested that ecdysone might control puffing
and hence transcription of the genes.
Anlmal Development
18.7 COMPARISION BETWEEN METAMORPHOSIS IN
AMPHIBIANS AND INSECTS
You may have realized that the metamorphic process in amphibians and insects show
certain fundamental similarity. Perhaps you have been able to identify them. They
are given below:
(1) In both insects and amphibians the stimulus necessary for the secretion of
hormone responsible for initiation of metamorphosis is provided by secretory
organs associated with the brain, namely hypothalamus in amphibians and
protocerebrum in insects.
(2) In both cases the hormones influence differentiation of cells and morphogenetic
processes during metamorphosis.
(3) In both amphibians and insects the brain and its secretion does not act directly
on the larval tissue, instead they stimulate the secretion of another endocrine
gland, the thyroid in amphibians and the prothoracic gland in insects.
(4) Metamorphosis in both involves some destructive activities (histolysis) and some
constructive activities (histogenesis).
(5) The juvenile hormone of insects however has no counterpart in amphibians and
so in amphibians there is no hormone which can check or prevent precocious
development.
(6) Larval growths in amphibians is not divisible into instars or inter stages.
Instead, it is continuous development. In insects, growth occurs periodically
marked by moults. There seems to be no growth in the intermoult period.
You can thus see that a comparison of metamorphosis between two groups which are
so widely separated phylogenetically as insects and amphibians and which have a
certain number of differences also reveal a striking similarity. The onset of puberty
in humans as you can read in Box. 18.1 also exhibit a striking similarity to
metamorphosis.

Box 18.1

Metamorphosis and Puberty in Humans

The hormonal basis of puberty in humans is very striking, and research suggests
that the process of puberty may in fact be qdite similar to metamorphosis. As
you have read, the metamorphoses of both amphibians and insects were both seen
to be regulated by hormonal changes that were initiated by the brain or
neurohormones (TSHRF and PTTH, respectively). Similarly in humans the
changes accompanying puberty in both sexes are initiated by luteinizing hormone
releasing factor (LHRF) from the hypothalamus of the brain (Fig.18.31). Like
TSH-RF, this factor is released from the hypothalamic neurons to the pituitary
gland. Likewise the LHRF is then transported by the blood vessels of the pituitary
to the anterior lobes of the gland. Once in the anterior pituitary, the releasing
facto~causes the release of a tropic hormone. In human puberty, the LHRF
releases the luteinizing hormone (LH) and the follicle stimulating hormone (FSH).
'These two hormones are collectively called gonadotropins as they stimulate the
development of ovaries in females and testes in males. Due to this stimulation,
the gonads secrete the sex hormones estrogen from the ovaries and testoterone
from the testes. The numerous morphological and behavioural changes of puberty
are thus due to the activity of these hormones on the various target tissues. --
As in metamorphosis, there appears to be in puberty a maturation inhibiting
hormone, whose activity decreases in order to permit the reactivation of
development. In humans this particular hormone is most probably melatonin,
whose serum concentration is found to decrease with the increase of LH
concentration. So you can see that puberty has many parallels to metamorphic
changes as it is a hormonally controlled reactivation of development, leading to
maturity and several changes in bodily form and physiology.
r -- -- r
 Male m d a r y
testis' sex hnraaeristics

ovary

I Fig. 18.31: BypoCb.l.so~HuHy-gamddub L man#mlhasexual- d

' J
SAQ 5
1) In one or two sentences, define the following terms:
a) stadipm b) instar c) imago. d) ExopterygOta
e) holmetabolous metamorphosis f) imaginal discs
ii) Fill in the blanks with suitable words
a) A .................................is necessary for initiating moulting in Rhodinus.
b) Exposure to .................................breaks the diapause in pupae of
Platysomia cecropia ' . '

c) Activation hormone secretkd from the ..........................of the brain and

stored in .............................................................
acts on prothoracic gland
which releases ...........................
d) The ...................... hormone released Erom corpora allata is a '

...........................compound.
e) ..........................
hormone is responsible for retention of larval characiers
and ecdysbn~promotes the ...............................process.
f) Removal of ............................ may result in precocious metamorphosis.
g) Ecdysone is converted into ............................ an active moulting hormone.

.18.8 SUMMARY
a After birth or hatching the young of an animal may develop directly into (he
adult form or the newly hatched individual may show indirect development.
appearing first as a larva morphologically and functionally and then
transforming into an adult by metamorphosis,
Larva occur in a number of most animal phyla ranging from porifora to the
vertebrates (anurans and some fishes).
Metamorphosis is one of the many ways in which an animal overcomes the
problem of growth, owing to the egg being essentially smaller than the adult.
In animals undergoing metamorphosis the larva which emerges after development
from the egg is smaller than the adult but may be (i) similar to the adult,
differing only in size and sexual maturity or (ii) greatly different from the adult
in habitat, fonn, organization, physiology as well as in sizq and sexual maturity.
In either case it transforms into an adult by the process of metamorphosis.
Metamorphosis is a phenomenon in which the larva changes in morphology,
organization and physiology before it transforms into an adult.
 Different animals show different degrees of changes during metamorphosis.
Thus (i) in urodeles and hemimetabolous insects where larva and adults are
largely similar, differing only in size and sexual niaturity and where other
changes are few of metamorphosis is gradual and incomplete and is called
incomplete metamorphosis. In animals like anuran amphibians and
holometabolous insects where the larva and adult show enormous differences,
metamorphosis is more radical and is called complete metamorphosis.
Metamorphosis is accomplished by cell .death, cell proliferation and cell
differentiation. Many genes active in larva switch off while many genes inactive
in the larva become active.
Metamorphosii is essentially controlled by hormones. But many extrinsic and
intrinsic factors influence metamorphosis, by intiating hormonal action.
In the amphibians the pituitary-thyroid hypothalamus axis control metamorphosis.
The main effecter hormone is thyroxin T4 a precursor of T3. Removal of the
thyroid prevents metamorphosis and addition of iodine or powdered thyroid gland
in water of the tadpole accelerate it. The thyroxine is secreted by the thyroid as
T3. This secretion is triggered by the production of thyroid stimulating hormone
(TSH) from the pituitary. The increased production of TSH causes
metamorphosis. Before the onset of metamorphosis high levels of prolactin (a
hormone which promotes growth and inhibits metamorphosis when level of
e thyroxin is low) and low level of thyroxin do not allow metamorphosis to take
place. The increasing levels of thyroid secretion cause the tadpole to undergo (i)
premetamorphosis (ii) prometamorphosis and finally when levels are high (iii)
metamorphosis climax. The high levels of thyroxin make the prolactin action
ineffective.
The response of the cells to any given hormone is inherent in them and it is
independent of the position of the cell in the body. Some cells undergo necrosis
while others proliferate under the action of the same hormone. Also cells of
the same organ but at different stages of development respond differently to the
same hormone.
n
During metamorphosis, in insects the neurosecretory cells of the brain secrete a
brain hormone (BH) or Activation hormone (AH). When BH which is stored in
corpora cardiaca is released as the active hormone prothoracicotropic hormone
P'ITH it stimulates the prothoracic gland to produce the moulting prohormone
ecdysone which changes into the active ecdysterone hormone. This stimulates
growth and moulting. During the larval life of insects the corpora allata of the
brain secretes juvenile hormone (JH) which suppresses metamorphosis aneach
moult. The quantity of JH decreases with successive moults and when its
concentration becomes low the larva develops into a pupa or into the last larva
stage (where pupa formation does not occur). When the JH is absent the p u p
or the last larva stage changes into the adult. The ecdysone appears to affect
mRNA synthesis (transcription) as is experimentally shown by changes in the
puffing patterns of polytene chromosome of insect. Moulting hormone also
alters K+ content in cell and nucleus.

18.9 TERMINAL QUESTIONS

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- -- - - -

1) Why is the marine environment more conducive for the developing larvae than
fresh water or terrestrial environment?

................................................................................................................
A................

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2) Briefly describe the process of metamorphosis in anwans.
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3 List the ~gressive/progressiveand constructive changes that occur during

amphibian metamorphosis.

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4) Briefly discuss the hormonal control of metamorphosis in amphibians.

5) #at are the molecular responses that the thyroid hormone evokes during
metamorphosis?
6) What is neoteny? Briefly discuss neoteny in different species of urodeles.
What is the significance of neoteny in evolutionary process?
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7) Distinguish the terms gradual and extensive metamorphosis with suitable
examples.

8) How do different hormones interact to bring about metamorphosis in insects?

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18.10 ANSWERS
Self Assessment Questions
1 i) Direct development relers to the type of development in which the new
born at the time of birlh resembles the parent except for the size and sexual
maturity. Direct development is observed in reptiles, birds and mammals.

ii) Indirect development is observed in many invertebrates, as well as in

protochordates like amphioxus of and also in vertebrates such as
amphibians. The young ones that hatches out of the eggs do not resemble
lhe adults, and infact appear very different from the parent. Such young
ones are called larvae which lead an independent life, subsequently undergo
transformation in heir structure by lhe process called metamorphosis and
then rese~nblelhe adults.
2 i) (a) False (b) False (c) True (d) True (e) True (f) True Metamorphosis

ii) a) v; b) iv; C)iii'; d) vi; e) ii; f) i

3 i) a) aquatic, terrestrial b) neoteny c) second d) activation, brain hormone
e) premetamorphosis, prometamorphosis, metamorphic climax f) regressive,
progressive, constructive

ii) a) progressive b) regressive c) regressive

d) constructive e) constructive f) constructive
g) constructive h) progressive i) regressive j) progressive
iii) 1) Switch over from ammonia excretion to urea excretion
2) Ability of the liver to synthesise enzymes of urea cycle.
3) Ability of kidneys to maintain water balance by increased water absorption.
4) Replacement of larval haemoglobin by adult haemoglobin.
iv) Progressive Changes:
1) Development of eye lid
2) Development of a multilayered and cornified skin

Regressive changes
1) Resorption of external gills and closure of gill clefts
2) Reduction of visceral skeleton
4 i) a) true b) false
' c) false d) true
e) m e f ) true
g) false h) true
i) m e j) false

ii) a) iii
c) iv
5.i a) Stadium: The interval between two moults during the larval period of an
insect is known as stadium.

b) Instar: Instar refers to the form an insect assumes as a result of moulting.

c) Imago: The final adult form that emerges at the end of metamorphosis in
an insect

d) Exopterygota: A division of subclass Pterygota of class insecta in which

the wings of the insects arise as a result of invagination of the integument.

e) Holometabolous metamorphosis: Metamorphosis of insects which produce

young ones that are completely different from their parents both in form
and habits.

f) Imaginal discs: Imaginal discs are the rudiments of future organs of the
adult such as wings antennae, and internal organs.
. .

2 ii. (a) blood meal (b) cold (c) protocereberum, corpora cardiaca, ecdysone
(d) juvenile, terpenoid (e) juvenile, moulting (f) corpora allata (g) ecdysterone.
Terminal Questions
1) See subsection 18.4.1
2) Seesubsection18.5.1
3) See subsection 18.5.1
4) See subsection 18.5.4
5) See subsection 18.5.7
6) See subsection 18.5.8 and 18.5.2
7) See subsection 18.6.2
8) See subsection 18.6.5.