MODULE 4

Activity 4.1
What do you understand by intercellular communication? give examples of intercellular
communication that occur in the body other than mentioned above.
Intercellular communication is the reaction or signaling between one cell to another to carry out
cellular function. Cells communicate with each other via direct contact (juxtacrine signaling), over
short distances (paracrine signaling), or over large distances and/or scales (endocrine signaling).
Why is intercellular communication important?
i The ability of cell to perceive and correctly respond to their microenvironment is the basis
development, tissue repair, immunity and normal tissue homeostasis.
i Important for evolution of organs and groups of specialized cell within organ (tissue),
regulation of proliferation, apoptosis, and differentiation of specialized cells.
i The cell signaling help to response to be in the right cells, at the right time, and in proper
coordination with the other cell.


If error happens, would lead to cancer, autoimmunity and diabetes. terutogenesls, neuroputhy, lnfertlllty.

How are cells able to receive and process information from the other cells?
Give exmples.:

1. Reception
i Target cell detect the signal from outside the cell by the binding action of the
signaling molecules to a receptor protein located at the cell surface
i Adrenal gland secretes epinephrine which binds at the surface of liver cell or the
hepatocytes in the liver which consist of G-protein at the plasma membrane.
2. Transduction
i The binding of the signalling molecules change the receptor protein which leads to
transduction (relay molecules.) Cellular response
i Cascade of event are triggered such as the synthesis of cylic AMP as second
messenger.
3. Response
i The transduced signal finally trigger specific cellular response.(such as glycogen
phosphorylase, rearrangement of the cytoskeleton, or activation of specific genes in
the nucleus.)
i cAMP leads to synthesize of enzymes for glycogen breakdown. The glycogen is
converted into glucose and releases in the bloodstream thus increase the blood
glucose level in the blood.






ACTIVITY 4.4. Routine cellular functions are also regulated via intercellular
communication.
1. Give two examples of routine cellular functions (refer to Activity 4.1).
Cellular respiration, protein synthesis

2. Explain how thyroxine, a hormone released from thyroid gland, and insulin and glucagon
released from the pancreas can influence cellular respiration in other cells.
Thyroxine is the body's main metabolic hormone, regulating the rate of oxygen use by cells.
Hence, this help in cellular respiration.
Insulin helps the cells take in glucose and convert it to energy. The energy will be stored at cells
and skeletal muscle. Respiratory muscles (skeletal muscle) like intercostals muscle and
diaphragm will contract due to presence of energy and hence undergo cellular respiration.


3. What kinds of hormones can influence protein synthesis? How is this related to intercellular
communication?
Steriod hormone can influence protein synthesis. Steroid hormones pass through the cell
membrane and bind to a specific steroid hormone receptor protein. This helps in
communicating the cell with steroid hormone and thus undergoes protein synthesis.

ACTIVITY 4.5: Disturbance in intercellular communication
You have explored the importance of intercellular communication in routine and specific
cellular functions. By giving specific examples and using schematic diagrams, explain the
possible causes of disturbance in intercellular communication? What would be the
physiological consequence?
One of the possible causes is defective in gap junction in the carcinogenic process. Connexins present at
gap junction act as conditional tumour suppressors (penindas, pengahalang) that modulate
(menyesuaikan, mengubahsuai) cell proliferation, as well as adhesion and migration. Means that
connexins restrict tumour growth. So, defect in gap junction or more specific in connexins, may lead to
carcinogenesis in which normal cells are transformed into cancer cells.

ACTIVITY 4.6. Types of intercellular junction
1. What are desmosomes?
Desmosome is a structure that forms the site of adhesion between two cells. It is a complex
disk-shaped structure that is matched with an identical structure at the surface of adjacent cell.
It also called macula adherens.

Draw a two-dimensional model of desmosomes between two adjacent cells.



Which cells are they found on?
Cells of the outer skin layer, heart muscle, and neck of uterus that are subjected to great
mechanical stress.

What is their function?
Provide a firm adhesion among cells

2. What are tight junctions?
Tight junction is an intercellular junction between epithelial cells in which the outer layers of the
cell membranes fuse. It is most apical intercellular junctions. It forms a band completely
encircling the cell. It also called zonula occludens.
Draw a two-dimensional model of tight junctions between two adjacent cells.


Which cells are they found on?
Cells that line the small intestine and kidney tubules.

What is their function?
Form a seal that prevents the flow of materials between epithelial cells in either direction

3. What are gap junctions?
Gap junction is a specialized intercellular connection between a large of animal cell-types. It can
occur almost anywhere along the lateral membranes.


Draw a two-dimensional model of gap junctions between two adjacent cells.


Which cells are they found on?
In adults, gap junctions are found in heart and smooth muscle cells

What is their function?
Allow cells to act in a coordinated manner


Activity 4.7 The blood brain barrier

The 600-km long capillaries in the brain (yes, that long because we have millions of them),
just like any other capillaries, are made up of endothelial cells, but these cells are attached
very firmly and closely together. Their membranes overlap to form a barrier of tight junctions
with no clefts and vesicles, unlike other capillaries. This shields delicate brain tissues from
toxins in the blood stream and from biochemical fluctuations, and allows selective drug
delivery to the periphery without affecting the brain cells. For example, some antihistamines
do not cause drowsiness because they cannot breach the blood-brain barrier.
y If you were a medical scientist trying to design a drug that can penetrate the blood-brain
barrier so that diseases like Alzheimers, Parkinsons, brain tumour, and AIDS relate brain
infections could be treated, how would you study and modify the gap junctions in the brain
capillaries? What would be the consequence of the modifications?

A mechanism that controls the passage of substances from the blood into the cerobrospinal
fluid and thus into the brain and spinal cord. The blood-brain barrier (BBB) lets essential
metabolites, such as oxygen and glucose, pass from the blood to the brain and central nervous
system (CNS) but blocks most molecules that are more massive than about 500daltons. This is a
low mass in biomolecular terms and means that everything from
hormones and neurotransmitters to viruses and bacteria are refused access to the brain by
the BBB. It also means that many drugs, which would otherwise be capable of treating disorders
of the CNS, are denied access to the very regions where they would be affective.





Channels are formed by pairs
of adjacent connexons formed
by 6 protein subunits.
Allow exchange of nutrients
and signal molecules between
cells without loss of materials
into the intercellular space.
Activity 4.8 Gap junction in cardiac muscle

Cardiac muscles are joined to each other by gap junctions where electrical impulses can flow
through. Contraction of cardiac muscle cells starts from the apex (the bottom part) where the
cells are stimulated by impulses coming from the pacemaker (AV-node) via the Purkinje
fibres. The contraction continues anteriorly in a synchronized manner due to the flow of
electrical impulses via the gap junctions. This ensures that the blood is pumped efficiently out
into the arteries.
y By referring to the histology of the cardiac muscle cells, please explain the importance of gap
junctions in the pumping action of the heart.
The action potential that drives contraction of the heart passes from fiber to fiber through gap
junctions.
Significance: All the fibers contract in a synchronous wave that sweeps from the atria down
through the ventricles and pumps blood out of the heart. Anything that interferes with this
synchronous wave (such as damage to part of the heart muscle from a heart attack) may cause
the fibers of the heart to beat at random called fibrillation.

y An alien from outer space is found to have no gap junctions in his heart. How does he
maintain the pumping action of his heart like a human being?






Activity 4. 9 chemical signalling via membrane molecules.

y Describe how specific membrane molecules take part in the process of growth and
differentiation of tissues. Give examples.

y Describe how specific membrane molecules take part in the functioning of cells that protect
the body against microbes and foreign cells. Give examples..
y One way in which T cells recognize foreign antigens is through the major
histocompatibility complex (MHC). For example, a foreign invader such as a
pneumococcal bacterium (which can cause pneumonia) may be engulfed by a
macrophage.
y The bacterium is processed in the macrophage in such a way that only a portion of the
bacterium is attached to the cell surface of the macrophage. On the surface of the
macrophage, this bacterial antigen is "presented" and bound as part of the MHC.
y Many types of MHC molecules exist; helper T cells specifically recognize antigens of
the MHCII type. The macrophage is just one cell type that may present antigens in the
MHCII complex; other cells, including B cells and dendritic cells, may also act as antigen-
presenting cells (APC).





ACTIVITY 4.10: Components of intercellular communication

y Using an example that you have already worked on in Activity 4.2-4.4 or another example of
your own:
Reabsorption of water in kidney tubule when the plasma osmolarity increases.

y Name the communicating cells
Cells of pituitary gland

y Identify the communicating signals
Releasing of hormone- ADH

y Identify the type of receptor on the target cell
Osmoreceptor

y Propose the transduction process
To transform extracellular message into intracellular signal or second messenger

y Explain the final response of the target cell to the communicating signal
The permeability of the kidney tubule increases for the water reabsorption.

y State the physiological effect of the intercellular communication
The plasma osmolarity inside the body decreases.


ACTIVITY 4.11: The communicating signals

y With the aid of your own diagram, compare and contrast between hormones, neurohormones,
neurotransmitters, and autocrine agents that act as communicating signals in terms of the cells
producing the agents, the target cells, action on the target cells, and the response of the target
cells.

Types Cells
producing
Target
cells
Action on
target cells
Response of target
cells
Hormones
(endocrine &
exocrine
hormone)
Beta cells Muscle
cells
Produce
insulin
Store glucose in the
form of glycogen
Neurohormones
(ADH)
Neurosecretory
cells in
hypothalamus
Epithaliel
cells of
kidney
tubule
Produce
ADH
Water reabsorption
Neurotransmitters
(acetylcholine)

Neuron Muscle cell Release of
acetylcholine
to the
synaptic cleft
Muscle contraction
Autocrine agents
(cell produce
compound self
use)
T cells T cells Binding to a
peptide,
MHC
complex on
APC
Macrophage
activation, B cell
activation and cell
mediated cytoxicity


y List four types of chemicals that function as first messengers. Give examples.

Hormones
(compound that acts at a distant site)
Neurohormones
(chemical that is released by neurons
that act at a specific distant sites)
Progesterone
Estrogen
LH
FSH
Insulin
throxine
TRH
Dopamine
Epinephrine
Neurotransmitters
(compound that binds to receptors
on neurons that either stimulate or
inhibit the action potential)
Autocrine agent
(compound that bind to the cell
surface receptors of itself)
Acetylcholine
Norepinephrine
Dopamine
GABA (gamma aminobutyric acid)
Glutamate
Serotonin
Endorphin
Cytokine interleukin-1



Activity 4.12. Characteristics of target cell response

If we hold the number of receptors
constant:

Why is the relationship between percent of receptors bound and the concentration of messenger not
linear? What would be the optimum condition? How can we apply this phenomenon in drug action for the
treatment of a disease?



- Because there is optimum condition (the concentration of the messenger)
- By increasing the no. of receptors required for the drug to attach.





If we hold the concentration of
messenger constant:

How would the relationship between number of receptors bound and the number of receptors be?


- Not linear.


For a specific number of receptors, the target cell response would also depend on the affinity of the
receptors to the messenger. What could increase and decrease the affinity?



Increase- greater intermolecular force between the ligand and its receptor

High affinity ligand binding implies that a relatively low concentration of a ligand is adequate to maximally
occupy a ligand binding site and trigger a physiological response

Decrease- low affinity ligand binding involves less intermolecular force between the ligand and its
receptor

Low affinity binding implies that a relatively high concentration of a ligand is required before the binding
site is maximally occupied and the maximum physiological response to the ligand is achieved


ACTIVITY 4.13

Name and draw a labeled schematic diagram of a target cell for the following first messengers:
Protein hormone (name the hormone)
Steroid hormone (name the hormone)
Neurotransmitter (name the neurotransmitter)
In or on the target cell, draw appropriate receptors for the above messenger molecules.
Binding of the first messengers to the receptors causes receptor activation. Propose specific
responses of the target cells as a result of receptor activation.



ACTIVITY 4.14

What is receptor activation?
- Binding of a ligand to a receptor activates the receptor, a process called receptor
activation.

What is the significance of the target cell final responses listed above in intercellular
communication? Give specific examples including those in homeostasis. Hint: refer to
Section 3.4 and 3.5.
Signal Type signal molecules cell or tissue origin Target cell Responds
Gas Nitrous oxide Vascular
endothelium
Smooth muscle
protiens Growth hormone Pituitary
somatotropes
various Secret growth
hormone for
growing
Interleukin I macrophages Lymphocytes Immunization
Insulin B cell of the
pancrease
Most body cells Secret insulin
to decrease the
glucose level
of body cells
Steroid aldosterone Adrenal cortex Renal tubular cells Secret
aldosterone to
increase the
blood pressure
progesterone Corpus luteal cells
and placenta
Uterine endometrium
and myometrium.
Thickening the
wall of uterus
cavity

Neurotransmitter acetylcholine Pre-synaptic
terminal
Post-synaptic terminal
- The significance of the target cell final responses listed above in intercellular communication
is to maintain the normally body function. For example the process of regulation of blood
glucose concentration. High blood glucose level detected by insulin-secreting cells of
pancreas, the B cell. Insulin secreting cells of pancreas stimulated to release insulin into the
blood. Most of the body cells take up more glucose and liver takes up glucose and stores it
as glycogen. The blood glucose level declines to a set point. Stimulus for insulin release
diminishes and body return to homeostasis.

In 2000, the Nobel Assembly at Karolinska Institutet has awarded the Nobel Prize in
Physiology or Medicine jointly to Arvid Carlsson, Paul Greengard and Eric Kandel for their
discoveries concerning "signal transduction in the nervous system". Carlsson discovered that
dopamine is a transmitter, Greengard found that dopamine and other "slow" transmitters act
by protein phosphorylation, and Kandel showed that phosphorylation is necessary for the
formation of short and long term memory.

Chronologically, signal transduction occurs after receptor activation and before the target
cell is able to produce the final response. Based on the works of the above scientists,
what do you expect to happen during signal transduction?

- Signal Transduction: ligand-receptor binding (termed receptor activation) causes
transformation of extracellular message into intracellular signal or second messenger.
How? Ligand binding leads to conformational change of the receptor or causes receptor
to interact with membrane or cytoplasmic enzymes/proteins. Consequence? Generation
of second messenger or activation of catalytic cascade that are transmitted to effector
molecules.

What are second messengers? Using examples, compare and contrast between second
messengers and signal molecules (first messengers).

- Many of the signal transduction processes involve second messengers.
They are intracellular substances that serve as relay from plasma membrane to
intracellular biochemical machinery, where they alter some aspect of cell function.
Second messenger is very important in prolong post-synaptic neuronal excitation or inhibition by
activating the second messenger chemical system.
The common second messengers include:
o Cyclic adenosine monohosphate (cAMP)
o Inosito triphosphate (IP3)
o Diacyl glycerol (DAG)
o Calcium ions (Ca2+)






ACTIVITY 4.15

List down the five general responses of target cells to signal molecules.Please describe specific
physiological examples for each of the five final responses of the target cells to signal molecules.
Remember to relate the examples to homeostatic regulation or to response to external stimuli.
ACTIVITY 4.15
FINAL RESPONSE
OF TARGET
CELLSFF
F
EXAMPLE POST-
RECEPTOR
PROTIEN
INVOLVED
EXAMPLES OF HOMEOSTASIS REGULATION
Alteration in the
permeability,
transport properties, or
electrical
state of the cells plasma
membrane.

Effect of
neurotransmitter
on
post-synaptic
neuron

Membrane
channel proteins
DD
acetylcholine (ACh) is produced by postganglionic
parasympathetic neurons (vagus
nerve) that impinge on cells of the SA-node in the
heart. When ACh receptor on the
membrane is activated, the cells respond by
slowing down heart rate, thus reducing blood
pressure.

Alteration in the cells
metabolism.

Effect of insulin
and glucagon on
glucose
metabolism.

Specific
intracellular
enzymes

High blood glucose level detected by insulin-
secreting cells of pancreas, the B cell. Insulin
secreting cells of pancreas stimulated to release
insulin into the blood. Most of the body cells take
up more glucose and liver takes up glucose and
stores it as glycogen. The blood glucose level
declines to a set point. Stimulus for insulin release
diminishes and body return to homeostasis.

Alteration in the cells
synthetic
and secretory activities.


Effect of glucose
on insulin
secretion in the
pancreatic -cell.

Proteins
(enzymes)
involved in
synthesis and
secretion

Same as above example.

Alteration in the cells
state of
proliferation.

Effect of growth
hormone.
Proteins
involved in cell
division

Growth hormone also called somatotropic hormone or
somatotropin. Growth hoirmone released by the
fibroblast. It cause growth of almost all tissues of body
that are capable of growing. It promotes increased sizes
of cells and increased mitosis, with development of
greater number of cells abd specific differentiation of
certain types of cells such as bone growth cells and early
muscle cells.

Alteration in the cells
contractile
activity.

Effect of
acetylcholine on
muscle
contraction.

Proteins
(troponin,
tropomyosin,
actin, myosine)
involved in
muscle
contraction

Nerves secret acetylcholine to open multiple
acetylcholine gated channels. this opening allow
sodium ions diffuse into the interior of muscle fibre
membrane and initial action potential. Action potential
stimulate sarcoplasmic reticulum to release Ca2+.
Calcium ion initiate attraction forces between the actin
and myosin filaments, causing them to slide along each
other, which is the contractile process.




ACTIVITY 4.16: The signaling events
1. Together.
2. Give a few examples that utilize the signaling events.
y Inflammatory responses helped by leukotrienes
y Gene transcription
y Sensory transduction
o olfactory signalling activation (vision) photoreceptors (rhodopsin G-protein coupled
receptor) hyperpolarization in response to light
o vestibular & auditory transduction hair cells bending the stereovilli of hair cells along
one axis causes cation channels to open/close

ACTIVITY 4.17: Mechanism of action of steroid hormones

Choose one steroid hormone.
y Glucocorticoid hormone

Name the organ that releases the hormone.
y Adrenal cortex

Name the target cell.
y general

Use the signaling events to describe in detail what happens after the hormone binds to the receptor in the
target cell.
1. Glucocorticoid hormone binds to GR (glucocorticoid receptor)
2. hsp90 dissociates from GR and exposes a nuclear localization signal that permits transport of GR
into the nucleus
3. receptor must remain hormone bound for receptor dimerization prerequisite for binding to GRE
on the DNA

What is the final response of the target cell and what is the physiological effect of this response?
y Dimerization leads to transactivation
y Physiological effect of this response body can cope with long term stress & raises blood
glucose level.



ACTIVITY 4.18: Receptors that function as ion channels


What ion channels are usually associated with the receptors?

y TSH, glucagon increases calcium channel & sodium channel
y Odorant increases adenylyl cyclise thus opens potassium channel

What are the first messengers for the ion channels?

y Chemical messengers odorants, chemicals that reflect metabolic status, ions, hormones,
growth factors, neurotransmitters.

What happens when these receptors are activated?

y The signal received at the cell surface is amplified and transmitted to specific sites within the cells
through 2
nd
messengers.
o Eg: hundreds of 2
nd
messengers can alter the activity of thousands downstream effectors.
y 2
nd
messenger systems allow specificity & diversity.
o Ligands that activate the same sognaling pathways in cell usually produces the same
effect.
o However, the same signalling molecule can produce distinct responses in different cells,
depending on complement of receptors and signal transduction pathways that are
available important to multicellular organism, eg: coordinated response of an organism
to a stressful situation.

Compare and contrast between ligand-gated Na+-channels and ligand-gated Ca
2+
channels. Relate to
final physiological responses.











- electrochemical driving
force for Na
+
is always
strongly -ve
- large inwardly directed net
driving force/gradient
favors passive movement of
Na
+
into every cell of body
(through open Na
+
channel)
- physiological use:
1. transmission of info
2. generate action potential
(eg: nerve impulse) in most
excitable cells.
3. found in epithelial cells
(eg: segements of renal
tubules & intestine) - Na
+
channels largely restricted
to apical surface of cell,
they allow Na
+
to enter cell
from the renal tubule
lumen instead. **key step
in mvment of Na
+
from
lumen to blood
Na
+
- electrochemical driving
force for Ca
2+
is always
strongly -ve
- Ca
2+
rapidly enters the cell
down an electrochemical
gradient once the Ca
2+
channels are open
- physiological use:
1. transmembrane signaling
for excitable & nonexcitable
cells.
2. generate action potential
in some excitable cells
Ca
2+
Using examples, relate the process involved when a message is transmitted by a cell via a first
messenger until the final response of the target cell is manifested. The events described must involve ion
channels. Mention the significance of the whole process






















ACTIVITY 4.19: RECEPTORS THAT FUNCTION AS ENZYMES

Do some research on insulin receptor on target cells (muscle cells and adipocytes).

Insulin facilitates entry of glucose into muscle, adipose and several other tissues. The only mechanism by which
cells can take up glucose is by facilitated diffusion through a family of hexose transporters. In many tissues -
muscle being a prime example - the major transporter used for uptake of glucose (called GLUT4) is made available
in the plasma membrane through the action of insulin.

What kind of proteins are these receptors made of?
y An insulin receptor is a tyrosine kinase, insulin receptors are an example of protein kinases.
y A kinase is an enzyme that catalyzes phosphorylation reactions, or reactions that add a
phosphate group to a substance by ATP.
y ATP, or adenosine triphosphate, is an organic compound that has three phosphate groups and
acts as an energy store for most organisms.
y In the case of insulin receptors, an amino acid called tyrosine that is found on other proteins is
phosphorylated

Odorant binds to specific olfactory
receptor protein in cell membrane
of a cilium in an olfactory receptor
Receptor activation stimulates
heterotrimeric G protein
subunit of G
olf
activates adenynyl
cyclise producing cAMP
cAMP binds to cAMP gated cation
channel
If receptor pot. Exceeds threshold, it
triggers action pot. In the soma that
travel down the axon and into the brain.
Opening of channel increases
permeability to Na,K,Ca
The net inward current
leads to membrane
depolarization and
increased Ca conc.
Increased Ca conc. opens Ca-activated Cl
-

channels produces more depolarization
bcs of high Cl
-
conc. Of olfactory receptor
neurones


What happens when insulin binds to the receptors? Briefly discuss receptor activation and
signal transduction.





















Insulin binds to a receptor on the plasma membrane of target cells, and when this insulin-receptor
complex forms, glucose is allowed to enter into the target cell where it is used as an energy source or
is converted into glycogen for energy storage
The insulin receptor is a protein consisting of two copies of two different peptide units,
alpha and beta. One insulin molecule is required to bind to each alpha subunit, and after
this occurs, the subunits then transmit a signal that causes the cytoplasmic end of the
receptor protein to change shape.
This change causes a cytoplasmic PROTEIN kinase active site to be exposed which causes
the phosphorylation of an insulin receptor substrate which initiates other reactions that
lead to the uptake of glucose by the cell
When the concentration of glucose in the blood is high, the pancreas releases insulin into
the bloodstream. The insulin travels throughout the bloodstream until it finds its receptors
on the surface of cells.
















What is the final physiological effect of insulin on the target cells? How does this come
about? Hint: focus on glucose uptake and glucose utilization (glycogenesis and fatty acid
synthesis) by the target cells.

When the concentration of glucose in the blood is high, the pancreas releases insulin into the bloodstream. The
insulin travels throughout the bloodstream until it finds its receptors on the surface of cells. After two insulin
molecules bind to the insulin-receptor complex, a series of complex reactions occurs which ultimately leads to the
uptake of glucose by cells. These cells are then able to use glucose as an energy source. When the concentration of
glucose in the bloodstream is low, most cells in the body use glycogen and fat as their energy source instead of
glucose.

What is the overall physiological effect of insulin in humans? What happens when there is
a lack of insulin or a deficiency insulin action? How do you treat these patients?
Insulin effect in humans is to maintain the blood glucose level in the blood. Diabetes mellitus is the
disease that lead to deficiency insulin action, which beta cells does not produce enough insulin or
does not produce insulin at all. Diabetes mellitus is treated by having insulin injections or drug
therapy.
ACTIVITY 4.20: Receptors associated with JAK kinase
Give an example of a receptor that are associated with JAK kinase and describe its
physiological role.
(Hint: cytokine receptor. How is this mechanism different from that of the membrane bound tyrosine
kinase?)
After two insulin molecules bind to the insulin-receptor complex, a series of complex
reactions occurs which ultimately leads to the uptake of glucose by cells. These cells are
then able to use glucose as an energy source. When the concentration of glucose in the
bloodstream is low, most cells in the body use glycogen and fat as their energy source
instead of glucose


JAKs, which have tyrosine kinase activity, bind to some cell surface cytokine receptors. The binding of
the ligand to the receptor triggers activation of JAKs.
With increased kinase activity, they form phosphorylate tyrosine residues on the receptor and create
sites for interaction with proteins that contain phosphotyrosine-binding SH2 domain.



ACTIVITY 3.21: G -proteins and adenylyl cyclise
Using the diagram in Fig. 4.13, describe the receptor activation and signal transduction processes when
an appropriate ligand binds to the receptor attached to Gprotein with the formation ofcAMP.



Explain the functions of cAMP and protein kinase A (PKA).



Give a physiological example that involves G-protein in production of cAMP in the transduction process.



ACTIVITY 4.22: G proteins and phospholipase C
y Using the diagram in Fig. 4.11, describe the receptor activation and signal transduction processes
when an appropriate ligand binds to the receptor attached to G-protein with the formation of IP3
and DAG.

Fig 4.11: Receptor coupled to a G protein, consisting of a single polypeptide chain with 7 membrane-spanning E
helical segments, an extracellular N-terminus, and a cytoplasmic C terminus.

Certain signalling molecule-receptor complexes activates G protein that activates enzyme
phospholipase C splits PIP
2
(a membrane phospholipid) (phosphotidylinositol-4,5-biphosphate),
into: IP
3
(inostiol triphosphate) & DAG (diacylglycerol)

DAG remains in plasma membrane combine with calcium ions activates protein kinase C
phosphorylate various target proteins i.e. growth, change in cell pH, regulation of certain ion
channels

IP
3
binds to calcium channels in ER causes release of calcium into cytosol


y Explain the functions of cyclic AMP and protein kinase C (PKC).

Second messengers ions or small molecules that relay signals inside the cell.

Cyclic AMP activates protein kinase C causes phosphorylation of protein in cell promote some
responses (cascade)

Protein kinase C one of the many types of protein kinase enzymes transduces the cellular signals
that promote lipid hydrolysis


y Give a physiological example that involves G-protein in the production of IP3 and DAG in the
transduction process.

Norepinephrine released by neuron and affect other neurons or effectors (Figure 4.15)


ACTIVITY 4.23: Receptors that function via G-proteins

The Nobel Assembly at the Karolinska Institute in Stockholm, Sweden, has awarded the Nobel Prize in
Physiology or Medicine for 1994 jointly to Alfred G. Gilman and Martin Rodbell for their discovery of "G-
proteins and the role of these proteins in signal transduction in cells".

y Discuss the significance of the above discovery. Give an example of a receptor that is associated with
G-proteins and describe its physiological role. How is this mechanism different from that mediated
by JAK kinase?

Significance because G proteins are found in many signal transduction pathways in animal cells
Example catecholamines receptors (Figure 4.15)



JAK kinase directed signalling activation of receptors will cause conformational changes

G-proteins - E-subunit of G protein sends signal to effector proteins cascade effects

y Based on Fig. 4.12, create animation to demonstrate the mechanism of action of a first messenger
mediated by a G-protein.

ACTIVITY 4.24: Production of second messengers

Illustrate how the four types of membrane receptors are involved in the production of second
messengers. Give appropriate examples.

y Receptors that functions as ion channels
i. Acetylcholine receptor binds to Ach allow sodium to enter result in transmission of
impulse & muscle contraction
ii. Ligand-gated chloride ion channels Gamma-aminobutyric acid (GABA) binds to chloride
channels open channel for chloride ions inhibits tramsmission of neural impulses

y Receptors that function as enzymes (protein kinase or guanylyl cyclase)
- Insulin (beta cell)

y Receptors that interact with cytoplasmic JAK kinase
Ras proteins

MAP kinases

y Receptors that interact with G-proteins
***refer 4.22



Activity 4.25 : cAMP (Cyclic adenosine monophosphate)

What is adenylyl cyclase? What is the substrate for adenylyl cyclase? What activates this enzyme?
Where in the cell is this enzyme found?
- Adenylyl cyclase is the enzyme that synthesizes cyclic adenosine monophosphate. The substrate
for adenylyl cyclase is adenosine triphosphate (ATP). It is activated by the attachment of a
hormone or neurotransmitter to a specific membrane-bound receptor. This enzyme can be
found in the plasma membrane (integral protein).

What is the name of the enzyme that is activated by cAMP? What happens when this enzyme is
activated? By using specific examples, explain the sequence of events that take place after this
enzyme is activated until a final physiological response is produced.
cAMP will activate an enzyme known as cAMP-dependent protein kinase (also termed protein kinase A).
when activated, protein kinase will add phosphate group (phosporylation) to the other enzyme.
Eg : regulation of glycogen
- Adenylate cyclase manufactures cAMP (cyclic AMP), which activates protein kinase A (cAMP
dependent protein kinase)
- This enzyme, in turn, activates phosphorylase kinase, which, in turn, phosphorylates glycogen
phosphorylase, converting into the active form called phosphorylase A.
- Phosphorylase A is the enzyme responsible for the release of glucose-1-
phosphate from glycogen polymers.



Activity 4.26 : Role of cAMP

Explain the sequence of events that involve second messenger cAMP in regulating cellular responses
to signaling molecules.
- Refer to the module.

Give physiological examples of cellular responses resulting from cAMP activities.
- used for intracellular signal transduction, such as transferring the effects
of hormones like glucagon and adrenaline, which cannot pass through the cell membrane.
- It also regulates the passage of Ca
2+
through ion channels.


In 1971, Earl Sutherland received the Nobel Prize in Physiology or Medicine for his work in elucidating
the role played by cAMP as a second messenger in signal transduction. In 1992, Edmond Fisher and
Edwin Krebs shared the prize for their part in demonstrating the role of protein phosphorylation in the
signal transduction process.
Discuss these important discoveries.
-

Activity 4.27 : Functions of IP
3
and DAG

Based on the diagram in Fig. 4.24, explain the sequence of events that involve second messengers IP
3

and DAG in regulating cellular responses to signal molecules.
- Refer to the module.

Give physiological examples of cellular responses resulting from IP
3
and DAG activities.
- IP3, generated from PIP2 has a vital role in the control of cellular and physiological processes as
diverse as cell division, cell proliferation, apoptosis, fertilization, development, behaviour,
memory and learning.


Activity 4.28: Ca2+ as second messenger
1. The concentration of Ca2+ in the intracellular fluid is far lower than that in the extracellular
fluid. Give the values. Describe how this phenomenon is maintained
y Within a typical cell, the intracellular concentration of ionized calcium is roughly 100nM,
but is subject to increases of 10100 fold during various cellular functions.
y The intracellular calcium level is kept relatively low with respect to the extracellular
fluid, by an approximate magnitude of 12,000-fold.
y This gradient is maintained through various plasma membrane calcium pumps that
utilize ATP for energy, as well as a sizable storage within intracellular compartments
y Calcium ion in ECF=8,4-10,5 mEq/L / 2.5mM
y Calcium ion in ICF =2-4 mEq/L / 1.5mM
y The active transport of calcium out of cytosol, via Ca2+ - ATPase, is one reason that the
cytosol of most cells has a very low calcium concentration compared to ECF. (120)

2. The increase in intracellular Ca2+ can be regulated and this is an important strategy to get a
physiological response from the cell. How is intracellular Ca2+ regulated? Give examples of
how Ca2+ is involved in modulating cellular physiological responses.
y 1. By means of active transport systems in the plasma membrane and cell organelles,
Ca2+ is maintained at an extremely low concentration in cytosol.
2. There is always a large electrochemical gradient favoring diffusion of calcium into
cytosol via calcium channels in both the plasma membrane and ER.
3. A stimulus to cell can alter this steady state by influencing the active transport
systems and/or the ion channels, resulting in a change in cytosolic calcium
concentration. (147)
y 1. Calcium binds to calmodulin. On binding calcium, the calmodulin changes shape,
which allows it to activate or inhibit a large variety of enzymes are protein kinase
2. Calcium combines with calcium-binding intermediary proteins other than calmodulin.
These proteins then act in a manner analogous to calmodulin.
3. Calcium combines with and alters response proteins directly, without the
intermediation of any specific calcium-binding protein.

Activity 4.29 Signal transduction via gene transcription
Compare and contrast between the effect on gene transcription by the lipid-soluble first messengers
and the lipid-insoluble first messengers. Give examples. (139-140)
y The receptors for these two broad chemical classes of messenger are in different
locations the former inside the cell and the latter in the plasma membrane of the cell
y Most lipid-soluble messengers are hormones steroid n thyroid
y Lipid-insoluble r mostly act as enzymes n ion channels (receptor)
y Lipid-soluble messenger function in the nucleus as transcription factors or suppressors
to alter the rate of transcription of particular genes
y Lipid-insoluble messenger function as ion channels and enzymes. Receptors that are
bound to and activate cytoplasmic JAK Kinases, and the receptors that activate G-
protein, which in turn act upon effector proteins either ion channels or enzymes in
plasma membrane.