PHYSIOLOGY OF EXCITABLE TISSUES

DR. EKEMENTEABASI ANIEBO UMOH

ARTHUR JARVIS UNIVERSITY

DEPARTMENT OF HUMAN PHYSIOLOGY
FACUL;TY OF BASIC MEDICAL SCIENCES
 CONCEPT CELL SIGNALLING
Cell signaling explains the communication between cells that are found in the human
body. The human body consist of over 75 trillions of cells. These cells has a task of
communicating with each other in a rapid way, delivering tremendous amount of
information in order to mediate a particular activity in the body. There are two basic
signals which are sent and received by cells during communication. These signals are:
1. Electrical signals and
2. Chemical signals
Electrical signals are changes in the cell’s membrane potential whereas chemical
signals are molecules secreted by cells into extracellular fluid. All of these signals are
activated by a particular cell with a view of passing an information or sending a message
to another cell. The latter cell which response to the electrical or chemical signal is
called the target cell. Many communications in the body is mediated via chemical
signals. However, in some cases, communication between cells might combine the
chemical and electrical signals. In chemical signal processes, there must be a receptor
on the membrane of the target cell. The receptor which are usually proteins and located
on the target cell binds to the ligand or molecule that was released into the extracellular
fluid. In cases where both chemical and electrical signals are used to communicate, the
binding of the receptor to the ligand can cause an alteration of the ion channels found
on the membrane of the target cells. This will subsequently change the membrane
potential of the target cell. Also, chemical signaling processes has certain
characteristics. One of such is specificity of the receptor. The receptors are specific to
respective ligands. This makes it difficult for such receptor to bind to any other ligand
that appears indifferent from the ones they know. Another characteristics is affinity of
the receptor. If a receptor has a strong affinity to a ligand, it binds to such ligand easily.
In some other cases, a protein or receptor can bind to several ligands. When several
ligands are released together at same time, there will be competition between the
ligands on the binding site of the receptor. The ligands in this case are called the
competitors. Hence, another characteristic of the chemical signaling process is
competition. Competing ligands are those which mimics each other. They are also
called agonist. Agonist may occur naturally or in synthetic forms. An example of a
natural agonist of acetylcholine is nicotine. Nicotine is found in tobacco and mimics
the activity of acetylcholine neurotransmitter. It binds to the same receptor of
acetylcholine. There also exist some antagonist which can bind to a protein or receptor
to decrease the activity of a certain ligand. Antagonist are inhibitory in nature. Both
antagonist and agonist are referred to as chemical modulators. Chemical modulators
are molecules that bind covalently or noncovalently to proteins and alter their binding
ability to a given ligand. Chemical modulators may activate or enhance ligand binding,
decrease binding ability, or completely inactivate the protein or receptor so that it is
unable to bind any ligand. Inactivation of receptors may be reversible or irreversible.
Generally, cell communication can be local or long distance communication. Local
communication occurs between nearby cells (short distance). There are basically three
methods in which cell passes information to each other in short distance or local
communication. These methods are:
 1. Gap junctions
2. Contact dependent signals
3. Autocrine and paracrine signals.
Long distance communication uses both chemical and electrical signals.

Short distance communication

Gap Junctions
Gap junctions usually allows cytoplasmic transfer of electrical and chemical signals
between adjacent cells. They are formed by proteins called connexins. These protein
arises from both adjacent cells and unites to form a path or channel called connexon.
These channels has the ability to open and close. When they open, the two adjacent cells
functions as a syncytium, behaving as if they were one cell with multiple nuclei. Gap
junctions allows for the passage of only small molecules such as amino acids, cyclic
adenosine monophosphate (cAMP), adenosine triphosphate (ATP) etc from one cell to
another. As such, larger molecules cannot pass through this junction. These junctions
are the only means through which electrical signals can pass directly from one cell to
another. However, movement of molecules or electrical signals can be modulated or
shut off completely following the activities of agonist or antagonist on the membrane
of the cells. Gap junctions has diverse isoforms and are located in various types of
human cells such as lungs, liver, smooth muscles, neurons and cardiac muscles etc.

Contact dependent signals

This type of cell signaling requires that the surface of one cell must make contact with
the surface of another cell. Hence, the surface molecule of one cell must bind to the
membrane protein of the other. It is applicable in the immune cells and nervous system.
In the nervous system, during growth and development, nerve cells can send out long
extension that must grow from the central axis of the cell body to the ends of a
developing limb.

Paracrine and Autocrine signals

Paracrine signals mediates their effects on cells that are within the vicinity of the
released paracrine molecules. Therefore, in this type of communication, there is a cell
capable of releasing a chemical molecule (paracrine molecules) into the extracellular
fluid. The paracrine molecules is targeted towards only close-by cells. It binds to their
membrane receptor and mediates its effect on the said target cell. Autocrine signals
mediates their effects on the cells that secretes them. Hence, in this communication, a
cell secrete a chemical molecule (autocrine) and the molecules in turn acts on the
secreting cell. Autocrine and paracrine cells reaches their target cells through the
interstitial fluid. Histamine is one example of a paracrine signaling molecule. Cytokines
and eicosanoids are also lipid-derived paracrine and autocrine signaling molecules.

Long distance communication

Long distance communication works through the chemical signal, electrical signal or
both of them. Long distance communication is carried out by the endocrine and nervous
system. Whereas the endocrine system works basically by releasing hormone (a
chemical substance) towards its target tissue, the nervous system works by both
electrical and chemical signals. In the endocrine system, a cell or gland has to release a
hormone in order to communicate with another. This hormones only binds to the other
cell if such a cell has its receptor. However, in the nervous system, an electrical signal
which is activated via a change in membrane potential travels along a nerve cell
(neuron) until it reaches the end of the cell, where it translate into a chemical signal
secreted by the same neuron. Chemicals secreted by neurons are called neurocrine
molecules. Neurocrine molecules can be a neurotransmitter, neuromodulator or
neurohormone. A neurotransmitter is a neurocrine molecule which is released from a
nerve into the extracellular fluid to have a rapid onset effect on the target cell to which
it was released. When neurocrine molecules acts slowly as an autocrine or paracrine
signal, it is called a neuromodulator. Also, when neurocrine molecules diffuses into
the blood for body wide distribution, it is called a neurohormone. It is the similarities
between neurohormones and classic hormones that bridges the gap between the
endocrine and nervosu system. These similarities causes the nervous system and
endocrine system to function together rather than act distinctly. Cytokines are one of
such molecules which functions to mediate both local and long distance communication
between cells. Though cytokines are involve in immune responses, they also function
in cell differentiation and development, a process that requires both long distance and
local communication. Some examples of cytokines include colony-stimulating factors,
growth factors, chemokines, inteferons, interleukins and tumour necrosis factor.

Signaling pathways: Components and features

Two components are essential in conveying an information from one cell to another via
chemical signals. They are

1. Ligand
2. Receptor

A ligand in this case refers to a hormone or even a drug. As a matter of fact, anything
that has the ability to bind on the receptor and mediate a change in the cell is referred
to as a ligand. However, in many cases, ligands are called hormones. It could be a
neurohormone, neurotransmitter or neuromodulator as the case may be, especially in
the nervous system. The classic hormones are the ligands in the endocrine system.
Whether a cell responds to the chemical signal sent by another cell or not will depend
on the receptor that is present on the target cell. A cell can only respond to a particular
chemical signal only if the cell has the appropriate receptor protein for such signal.
Therefore, in cases where the target cell has the receptor protein for the binding of the
ligand, a response will be initiated in the target cell and the information will be
successfully transferred or communicated. However, it is important to know that all
signaling or communication pathways shares the following features/stages:
 1. The signal molecule (ligand) binds to a protein receptor. The ligand is also called
the first messenger because it brings information to the target cells. In cases
where it has to activate another molecule in order to pass the information further,
the second molecule in the chain of communication is called a Second
messenger.
2. Binding of ligand to receptor activates the receptor.
3. The receptor in turn activates one or more intracellular signal molecules of which
the second messenger is a part as earlier stated in number 1.
4. The last signal molecule in the pathway creates a respond by modifying existing
protein or initiating the synthesis of new proteins.

Receptors: Location and Classification

As earlier emphasized, protein receptors are very essential in the transmission of
information from one cell to another. Protein receptors for signal molecules are key in
physiology and medicine. This is because many medications (drugs) are created to
target these receptors. These medications acts like the ligand or chemical signals and
binds to these receptors to mediate their effects. Receptors could be found or located in
the:

1. Cell membrane as integral proteins

2. Nucleus
3. Cytosol

Hence, information could be processed at the level of the nucleus, cytosol and plasma
membrane. Information processed at the nucleus level can take a while than those at
other levels. Also, the binding location of a chemical signal or molecule depends on the
nature of such molecules. Molecules can be lipophilic (molecules that can enter into
the cell through simple diffusion) or lipophobic (molecules that are unable to enter the
cell through simple diffusion) in nature.

Lipophilic signal molecules binds to cytosolic or nuclear receptors because they can
easily penetrate the plasma membrane. When once they binds to these receptors and
activates them, they turn on a gene and directs the nucleus to make new messenger RNA
(mRNA), a process called transcription. The mRNA provides a template for synthesis
of new proteins, another process which is simply called translation. This kind of
communication as seen in the nucleus takes hours or longer time. At the end of each
communication, there should be a turn off of the activated receptor. In cases where there
is no turn off of the receptor, a disease condition can set. Diseases such as cancer occurs
due to the inability of a receptor to turn off after responding to cell growth and
differentiation signals from a ligand. Hence, the cells keep growing and never stops.
Again, lipophilic signal molecules can also bind to receptors on the cell membrane
irrespective of the fact that than easily cross the cell membrane barriers. Emphasis for
binding depends on the location of their receptor.

Lipophobic signal molecules are confined to the extracellular fluid since they cannot
penetrate the cell membrane through simple diffusion. Hence, they usually bind to the
receptor protein found on the cell membrane. The response of target cell or time limit
to processed information that is communicated via chemical signals and cell membrane
bound receptors is very rapid. It takes place in milliseconds to minutes.

Membrane receptors are grouped into four major categories namely:

1. Receptor channel
2. Receptor-enzymes and
3. Integrin receptor
4. G-protein coupled receptors (GPCRs)

Receptor channels are the simplest form of receptors. As a matter of fact, they are the
chemically gated or ligand gated ion channels. The binding of a ligand or signal
molecules to these receptors causes either an opening or closing of their respective ion
channel. This eventually alters the ion flow across the membrane.

Receptor-enzymes are those receptors which activates an enzyme whenever a ligand

binds to it.

Integrin receptor are those receptors which will alter enzymes and cytoskeleton when
a ligand binds to it.

G-protein coupled receptors (GPCRs) opens an ion channel and alters enzymes
activity whenever a signal molecule or ligand binds to it. They are a major focus in the
concept of cell signaling because many functions in the human body are carried out
through these receptors. Therefore in signal transduction (The process of transmitting
information from one side of the cell membrane to the other, in order to initiate a
response), G-protein coupled receptors takes the central focus. Signal transduction will
be explained in details in a later part of this chapter.

The GPCRs helps to relate information from special kinds of proteins called guanine
nucleotide-binding regulatory protein (G-protein) to the cell. During signal
transduction, whenever a ligand binds to GPCRs, its effect will be mediated through G-
proteins to the target cells. Therefore, in a given cell, G-proteins are usually anchored
by G-protein coupled receptors (GPCRs). Signal transduction, protein transport, growth
regulation and polypeptide chain elongation are all mediated through G-proteins to the
respective cells that anchors them.

The GPCR is a seven (7) transmembrane loop which anchors the G-protein at the end
of the loop beneath the cell membrane. It is more like a wire that transverse the plasma
membrane of the cell from the exterior, entering in and out of the cell seven consecutive
times. The end of the wire beneath the plasma membrane attaches itself to the guanine
nucleotide binding regulatory protein (G-protein) which helps to pass on the
information to the cytosol. Figure 13. 1 shows the structure of the GPCR with the G-
protein attached to it.
Guanine nucleotide binding regulatory proteins (G-proteins): Regulation
and Classification

Regulation of G-proteins
The G-proteins has three (3) subunits which are closely bound together prior to the
binding of a ligand to their respective receptors (GPCRs). The three sub-units are:

1. Alpha(α) sub-unit
2. Beta (β) sub-unit and
3. Gamma (ɣ) sub-unit

Therefore, in the inactive state of the G-proteins i.e when no ligand binds to the
receptor, the alpha, gamma and beta subunit are closely bound together. However, the
gamma (ɣ) and beta (β) sub-units of the G-protein are more closely bound together and
functions together while the alpha (α) functions as a single unit. Hence, whenever a
ligand binds to the GPCR, the G-protein is activated and its alpha (α) sub-unit (Gα)
splits from the gamma and beta sub-unit (Gɣβ). This mechanism of activation is
anchored by an exchange between two substrates namely:

1. Guanosine Diphosphate (GDP) and

2. Guanosine Triphosphate (GTP)

Prior to activation when the G-proteins are in their inactive state, the Gα subunit is
bounded to guanosine diphosphate (GDP). However, during signal transduction, when
the ligand binds to the GPCR, the Gα exchanges guanosine diphosphate (GDP) for
guanosine triphosphate (GTP). This exchange is made possible by some guanine
nucleotide exchange factors (GEFs). As soon as the exchange is facilitated by the
GEFs, the G-protein splits into the alpha sub-unit (Gα) and the gamma/beta sub-unit
(Gɣβ) mediating the effect that was intended by the ligand which bounded to its receptor
on the cell. The two sub-units (Gα and Gɣβ) signals to different pathways following
activation. At several occasions, the G-protein doesn’t mediate the intended effect of
the ligand directly but rather works by activating some second messenger molecules
which will be discussed in a later part of this chapter. After the intended function or
effect is achieved, the sub-units of the G-protein are re-united by the activity of a
GTPase enzyme. This enzyme is an inherent property of the Gα sub-unit of G-protein.

Figure 13.1: G-protein coupled receptor (GPCR) and G-protein

The ligands that binds to GPCRs include hormones, growth factors, olfactory
molecules, visual pigments and neurotransmitters etc. Usually, the two sub-units of G-
proteins (Gα and Gɣβ) has the inherent ability to reunite immediately after the intended
effect and function has been established in the cell due to the enzyme GTPase. However,
in pathological conditions, these sub-units do not re-unite and so, the signaling pathway
of the G-proteins continues uncontrollably without an end. Drugs that are created in
such case are therefore targeted at the GPCRs to put an end to such situation. A good
example of cases where the sub-units of the G-protein do not get to re-unite is that of
Cancer. In cancer cases, where ligands such as growth factors binds to the GPCRs,
there is a switching on or activation of the G-Protein such that the Gα splits from the
Gɣβ. These two sub-units goes into different pathways to activate several other small
G-proteins. These pathways subsequently results in growth and proliferation of the
cells. Since the G-proteins can no longer switch off or re-unite its two distinct sub-units,
the growth and proliferation of such cell continues and never ends.

Classification of G-proteins
The G-proteins are mainly classified based on their alpha sub-unit (Gα). However, this
does not rule out other classifications that uses the gamma and beta sub-units of the G-
protein. The family of G-proteins following the alpha unit classification are as follows:
1. Gαs
2. Gαi
3. Gαq
4. Gα12
Each of these families of the G-protein are responsible for certain functions in the
human body. Hence, the G-protein that is involved in mediating neuroendocrine
functions will certainly not be the one involved in hematopoietic functions and so on. It
is therefore important to know the exact G-protein family involved in mediating a given
physiological functions. Again, each family of the G-protein can further be divided into
several other types in such a way that each of the types has their respective functions.
Gαs Family
This family is made up of the Gαs and Gαolf. S in this case stands for stimulation whereas
olf represents olfaction. Gαs is ubiquitous in nature having relevance in every part of the
body whereas Gαolf is expressed in olfactory sensory neurons. Signaling pathways
associated with Gαs family of the G-protein has been linked to several physiological
and pathological functions such as Myocardial hypertrophy, increase in hydrolysis of
triglyceride, decrease in amino acid uptake, increase in the conversion of glycogen to
glucose, inhibition of synthesis of glycogen, increase in synthesis of estrogen and
progesterone, increase in synthesis of aldosterone and cortisol, increase in
excitation/contraction and sympathetic cardiac activation and hypertrophy, iodide
organification, secretion of thyroxine, thyroid cell mitogenesis, increase in reabsorption
of calcium from bone, fluid secretion, inhibition of platelet aggregation and secretion.
However, there are other functions of Gαs family that has not been linked to well-
established signaling pathways. These functions include magnesium ions (Mg2+ uptake)
and hematopoietic stem cell engraftment in bone marrows.
Gαi Family
This is the largest and most diverse family of the G-proteins. i in this case represents
inhibition. Table 13.1 shows the members of the Gαi family and their respective location
in the body. Some members of this family are widely distributed in the body, others are
found everywhere in the body (ubiquitous) whereas others are found in specific areas
of the body.
Physiological and pathological functions that could be explained by signaling pathways
associated with the Gαi family of the G-protein include Vision, taste, vomeronasal
function, Cardiac activation (contractility), regulation of cardiac L-type Ca2+ channels,
leukaryote activation and migration, hepatic authophagy, developmental processes,
lipid metabolism, regulation of immune cells, renal function, platelet activation,
chemokine-induced lymphocyte migration. Other functions associated with this family
irrespective of the fact that the signaling pathways are not well defined include
transformation of fibroblast, spindle positioning during cell division, regulation of
diacylglycerol (DAG) kinase, neurotransmitter release in synapse and cell migration.

Table 13.1: Family of Gαi and their location of expression

S/N Gαi FAMILY LOCATION
1 αi1 Widely distributed in the body
2 αi3 Ubiquitous (found everywhere in the
body)
3 αi2 Widely distributed in the body
4 α0A Neurons
5 α0B Neuroendocrine
6 αt1 (t stands for transducin) Rod cells of the retina and taste cells
7 αt2 (t stands for transducin) Retinal cone cells
8 αg (g stands for gustducin) Taste and brash cells
9 αZ Neurons and platelets

Gαq Family
The Gαq consist of αq, α11, α14, α15 and α16. αq and α11 are ubiquitous in nature
whereas α14 is found in the kidney, lungs and liver. α15 and α16 are found in the
hematopoietic cells. There are several physiological and pathological functions of the
Gαq family with well-established signaling pathways. These functions include
myocardial hypertrophy, smooth muscle tone, platelet activation, hormone release in
anterior pituitary and synaptic transmission at Purkinje cell synapse. Other functions
that are linked to this family of G-protein but do not have well-established signaling
pathways include insulin secretion by pancreatic beta cells, leukocyte migration and
activation, embryonic myocardial growth, neural crest development and transformation
of fibroblast.
Gα12 Family
This family consist of α12 and α13. The two are ubiquitous in nature. This family
mediates several physiological and pathological functions with well-defined signaling
pathways such as platelet activation, smooth muscle contraction, leukocyte migration
and neuronal axon guidance. Other functions that are linked to this family of G-proteins,
though without a well-established signaling pathway include leukocyte activation and
proliferation, lymphocyte development, embryonic development of blood vessels and
angiogenesis, transformation of fibroblast, cancer cell invasion and metastasis.
Beta (β) sub-unit expression of G-proteins (Gβ)
The beta sub-units of the G-protein are mainly made up of five isoforms or genes (β1 –
β5). β1 – β4 are widely distributed in the body whereas β5 finds its place mainly in the
brain. β1 can also be found in the rod cells of the retinal cells while β2 are also found in
the retinal cones.
Gamma (ɣ) sub-unit expression of G-proteins (Gɣ)
The gamma sub-units of the G-protein has 12 genes isoforms (ɣ1 - ɣ5 and ɣ7 - ɣ13) in
human body. ɣ5, ɣ7, ɣ10, ɣ11 and ɣ12 are widely distributed in the body whereas ɣ2 is
ubiquitous. ɣ8 and ɣ9 are found in the olfactory epithelium, ɣ4 is found in the brain and
other tissues, ɣ3 is domicile in the brain, ɣ1 is found in the retinal rods and brain
whereas ɣ13 is mainly found in the brain and taste buds.

The large and small G-proteins

The large G-proteins are the ones earlier discussed above. They have the three sub-units
(αɣβ) which are bound together before activation. Following activation of these proteins
as ligand binds to their receptor (GPCR), the Gα exchanges GDP for GTP through the
action of guanine nucleotide exchange factors (GEFs). This exchange causes the G-
protein to split into the Gα and Gɣβ sub-units. The two subunits goes ahead to effect
the intended functions of the ligand that bounded the GPCR through certain pathways.
For every pathway, there is usually subsequent activations of some other small proteins
also known as the small G-proteins. These small proteins are activated in the pathways
of the Gα or Gɣβ after activation of the large G-proteins.

The structure of the small G-protein differs from that of the large G-proteins. Unlike
the large G-proteins which has two heterotrimeric sub-units (Gα and Gɣβ), the small G-
proteins has only one α-sub-unit. However, their activation is similar to that of the large
proteins. In their inactive state, the small G-protein alpha (α) sub-unit is bound to
guanosine diphosphate (GDP). Following the binding of a ligand to the GPCR, there is
a subsequent activation of the small G-protein. This involve the exchange of guanosine
diphosphate (GDP) with guanosine triphosphate (GTP) by guanosine nucleotide
exchange factors (GEFs) in the alpha (α) sub-unit of the small G-protein. The activation
of the small G-protein will result in subsequent downstream signaling process in order
to mediate the function by the ligand that bound to its receptor. Also, the activity of the
small G-protein is terminated by the GTPase-activating proteins (GAPs). GAPs
activates the GTPase enzyme which causes GTP hydrolysis. This will eventually cause
the alphas sub-unit to be bound once again with GDP. There are several types of small
G-protein. These types include the Ras superfamily and others such as Rho, Rab, Rac,
Sarl/Arf and Ran homologs etc. The Ras family consist of Ras, Ral, Rit, Rap, Rhep
and Rad. The Ras family is generally responsible for cell proliferation, Rho for cell
morphology, Ran for nuclear transport and Rab as well as Arf are responsible for
vesicle transport. The figure below (Figure 13.2) shows the activation of small G-
protein of the Ras family.

Figure 13.2: Activation of small G-proteins using Ras family as an example

Signal transduction and second messenger systems

Signal transduction explain the process which an extracellular signal molecule
activates a membrane receptor which in turn alters the intracellular molecules to create
a response. In this scenario, the extracellular molecule or ligand is the first messenger
whereas the intracellular molecule is the second messenger. The membrane receptor
acts as a transducer in this process, converting the message from extracellular molecules
into intracellular messenger molecules that eventually trigger a response. Hence, the
second messenger systems is activated following the binding of a ligand to the receptor
and they occur in the interior aspect of the cell (intracellular). Basically, signal
transduction follows the pattern below:

1. Binding of first messenger molecules or ligands to membrane receptor such as

GPCR.
2. Activation of receptor
3. The activated receptor turns on or switch on its associated protein such as G-
protein in a case where the receptor is a GPCR
 4. The switching on of the associated protein of a membrane receptor turns on the
second messenger cascades or system.

Second messenger cascade or system

The second messenger cascade or system that is activated by the proteins of the
membrane receptor has several components. The last component of the second
messenger system is what eventually brings about the response or effect that was
intended by the ligand which earlier bind to the membrane receptor on the cell. The
following explains the second messenger system appropriately:

1. The second messenger system or cascade is made up of second messenger

molecules which could be nucleotides, lipid derived substances or ions. The
system or pathway is often named according to these second messenger
molecules (nucleotides, lipid derived substances or ions). The various second
messenger molecules/pathway include:
A. Cyclic adenosine monophosphate (cAMP): cAMP belongs to the
nucleotide class of second messenger molecules. cAMP is synthesized or
made from adenosine triphosphate (ATP).
B. Cyclic guanosine monophosphate (cGMP): cGMP is also of the class of
nucleotide. cGMP is synthesized from guanosine triphosphate (GTP)
C. Inositol 1,4,5-triphosphate or inositol trisphosphate (IP3): IP3 is a lipid
derived second messenger molecule. IP3 is synthesized from membrane
phospholipids.
D. Diacylglycerol (DAG) also belonging to the lipid derived class of second
messenger system. DAG is also synthesized or made from membrane
phospholipids
E. Calcium ions (Ca2+) which belong to the ionic class of second messenger
system. Calcium ions occurs naturally in the intracellular and extracellular
fluid of the cell. It is also abundant in the sarcoplasmic or endoplasmic
reticulum of the cell.

N/B: IP3, DAG and Ca2+ are second messengers in the phosphoinositol signaling
pathway. Hence, they are often referred to as the phosphoinositol signaling pathway

2. The second messenger systems can activates enzymes which transfer a

phosphate group from ATP to a protein. These enzymes are called the protein
kinases. They include protein kinase A (PKA), protein kinase C (PKC), protein
kinase G (PKG) and Rho kinase. Sometimes, the second messenger are explained
or classify based on these enzymes.
3. Again, the second messenger systems contains some amplifying enzymes.
These enzymes are the ones responsible to create or activate the second
messenger molecules (cAMP, cGMP, IP3 and DAG) listed above. The
amplifying enzymes is not applicable in calcium signaling pathway as calcium
occurs freely in the ICF, ECF or intracellular stores (endoplasmic or
sarcoplasmic reticulum). Generally, the role of amplifying enzymes in the
system is to increase the effect of the ligand by activating the second messenger
 molecules. The second messenger molecules goes ahead to further activate other
enzymes and possibly alter (open) some ionic channels. This is the reason it is
called an amplifying enzyme. Some examples of amplifying enzymes are
adenylyl cyclase, guanylyl cyclase and phospholipase C.
4. There exist a form of relationship between the different second messenger
molecules/pathways. Many at times activation of the cGMP, cAMP and
DAG/IP3 pathway can increase calcium ion concentrations in the cell.

Cyclic adenosine monophosphate (cAMP) signaling pathway

The cAMP signaling pathways has the following components:

1. Ligand
2. G-protein coupled receptor (GPCR)
3. Adenylyl cyclase enzyme
4. Adenosine triphosphate
5. Protein kinase A
6. Cyclic adenosine monophosphate nucleotide (cAMP)

In the pathway, a ligand or first messenger signaling molecules such as hormone,

growth factor, olfactory molecule, visual pigment or neurotransmitter binds GPCR on
the specific plasma membrane of a target cell. The binding of ligand activates G-protein
couple receptor (GPCR). Since GPCR are linked to specific G-proteins, activation of
GPCR will subsequently switch on the G-proteins. There exist the splitting of the alpha
(α) sub-unit from the beta/gamma (ɣβ). These two sub-units enters into different
pathways to mediate a specific function. However, the function that is so mediated by
these sub units of the G-protein is via the activation of adenylyl cyclase enzyme. This
enzyme is capable of using adenosine triphosphate (ATP) found in the cell to
synthesize or make cyclic adenosine monophosphate (cAMP). The present of cAMP
will in turn activate protein kinase A (PKA). PKA are the last substrates in the cAMP
pathways. They initiate the response intended by the ligand on the target cell through
their ability to open certain nucleotide ion channels and further alter the membrane
potentials of such target cells. PKA has a localized and specific characteristics in a cell.
These characteristics is what ensures that their effect is targeted towards a certain
subcellular location. Compartmentalization of signaling or regulation of the location,
duration and frequency of signals from the cAMP pathway by PKA is a function of the
A-Kinase anchoring proteins (AKAP). AKAPs plays a functional role of tethering
PKA holoenzyme to specific subcellular locations in physical proximity to PKA targets,
increasing the specificity and efficiency of the cAMP signaling pathway.

This pathway is involve in regulation of several cellular functions in:

1. Cardiovascular system: Heart rate which is often as a result of contraction and

relaxation cycles of the cardiac muscle is regulated by protein kinase A. PKA
phosphorylates several proteins that are involve In excitation-contraction
coupling such as L-type calcium channel, ryanodine receptor (RYR), Troponin-
I, myosin binding protein C etc. They are also involve in regulating
 phospholamban which in turns regulates the activity of SERCA 2, resulting in
the increase uptake of calcium ion intake by Sarcoplasmic reticulum.
2. Steroid biosynthesis: cAMP plays a regulatory function on adrenocorticotropic
hormone (ACTH), follicle stimulating hormone (FSH) and luteinizing hormone
(LH). These three hormones are involve in acute steroid biosynthesis. CAMP is
also involve in cholesterol release from lipid droplet and cholesterol transport
across mitochondrial membrane. This function of cAMP is a rate limiting step in
providing substrate for cholesterol side-chain cleavage enzyme, a necessary
factor in steroid biosynthesis.
3. Regulation of reproductive functions: The initiation and maintenance of sperm
motility are thought to be involve cAMP dependent phosphorylation by PKA.
4. Regulation of metabolism in adipocytes: Lipolysis is a process increased by
PKA mediated phosphorylation of hormone sensitive lipase on adipocyte.
5. Regulation of exocytosis: Secretion of hydrochloric acid by parietal cells
through Hydrogen-Potassium ATPase is regulated by cAMP/PKA, insulin
secretion by beta cells of pancreas is regulated by reversible phosphorylation of
the beta cells where phosphorylation of PKA enhances insulin secretion and
protein phosphatase inhibits it while water absorption in the principal cells of the
renal tubule is also regulated by cAMP.
6. Regulation of immune functions: Prostaglandin 2 (PGE 2) and other ligands
elevating cAMP by binding to G-Protein coupled receptors inhibits T-cell
antigen receptor (TCR) induced T cell activation, thereby exerting important
immunoregulatory function through the activity of PKA.

Figure 13.3: cAMP signaling pathway

Cyclic guanosine monophosphate (cGMP) signaling pathway
Cyclic guanosine 3’5 monophosphate (cGMP) has two distinct pathway that regulates
its synthesis. Following the binding of a ligand to a receptor, cGMP is synthesized from
guanosine triphosphate (GTP) by an enzyme called Guanylyl cyclase. This enzyme
activates two distinct pathways of which one is coupled to natripeptide hormone and
the other is coupled to a single gas called Nitric oxide. There are two distinct forms of
guanylyl cyclase namely: Soluble guanylyl cylcase (SGC) which is responsible for the
synthesis of cGMP from GTP through the pathway coupled to nitric oxide and
Particulate guanylyl cyclase (PGC) which synthesizes cGMP from GTP through the
pathway coupled to natripeptide hormone. The SGC is a protein found in the cytosol
whereas the PGC is found on the membrane of the cell. However, intracellular
localization of the guanylyl cyclase is more expressed than on the membrane.
Following the activation of cGMP, its action is mediated by three types of effectors
namely: cGMP –gated ion channels, protein kinase G (PKG) and
phosphodiesterase (PDE). Whereas, PKG and PDE are predorminant in the
cardiovascular system, the ion channels are found in the rectinol and olfactory
neuroepithelium and neurons.

These signaling pathway is involved in activities such as cell proliferaton and increase
permeability, ion channel conductance, cell growth, apoptosis, cellular morbidity
and contractility. In the vascular endothelium, activation of protein kinase G through
the cGMP will result in cell proliferation and increase permeability; in the vascular
smooth muscles, it inhibits cell proliferation and mediates vascular relaxation whereas
in cardiac myocardium, it inhibit hypertrophy and modulates contractility. Apoptosis is
mediated in all these tissues following the cGMP signaling pathway.

Phosphodiesterase are often linked to crosstalks in signaling pathways of cGMP and

cAMP. Oftentimes, inhibition of the phosphodiesterase which are about eleven (11) in
numbers can enhance the activities mediated by signaling pathways of Cgmp while
inhibiting those of the cAMP.
 Figure 13.4: Nitric oxide/cGMP signaling pathway

Phosphoinositol (IP3, DAG and Ca2+) signaling pathway

IP3, dag and Ca2+ are second messengers molecules of the phosphoinoditol signaling
pathway. As found in other pathways, a ligand or primary messengers such as
epinephrine, acetylcholine, and hormones such as GnRH, GHRH, ocytocin etc binds to
their respective G-protein coupled receptors. In all cases, epinephrine often binds to α1
GTPase protein coupled receptor (GPCR) and acetylcholine binds to M1 and M2
GPCR. The conformational change of GDP exchange for GTP will be the result in the
α1 receptor following the binding of the ligand to GPCR. When once the α1 is activated,
there will be subsequent activation of phospholipase C which will hydrolyzes the
membrane bound phosphatidylinositol 4,5 biphosphate (PIP2). Hydrolyzing PIP2
often results in the formation of secondary messengers DAG and IP3. IP3 binds to
calcium pump on the endoplasmic recticulum, transporting calcium, which is another
second messenger into the cytoplasm. Calcium ultimately binds to many other proteins,
activating a cascade of enzymatic pathways.

Figure 13.5: Phosphoinositol signaling pathway

This signaling pathway is involved in involve in excitation-contraction coupling of
smooth muscles, insulin secretion, exocytosis, proliferation and lymphocyte
functions etc.