DEPARTMENT OF HUMAN PHYSIOLOGY,

FACULTY OF BASIC MEDICAL SCIENCES,

COLLEGE OF HEALTH SCIENCES, AHMADU BELLO
UNIVERSITY, ZARIA.

ADVANCED NERVE AND MUSCLE PHYSIOLOGY

(HPHY 811)

THE PATHWAY AND FATE OF SYNAPTIC VESICLES

JULY, 2019
1
 VESICULAR PHYSIOLOGY
INTRODUCTION
 A vesicle is a small structure within a cell, consisting of liquid or
cytoplasm enclosed by a lipid bilayer membrane.

 Golgi apparatus and ER also form vesicles that play a role in

secretion.

 Membrane-bound cytoplasmic vesicles are either secretory vesicles

that contains proteins that will be released from the cell (e.g synaptic
vesicles) or storage vesicles such as lysosomes and peroxisomes never
leave the cytoplasm.

 Molecules such as hormones and neurotransmitters synthesized by the

cell are transported around the cell in vesicles.
2
VESICULAR PHYSIOLOGY
INTRODUCTION
 Vesicles can fuse with the plasma membrane to release their contents
outside the cell.

 Vesicles can also fuse with other organelles within the cell
(Silverthorn, 2009).

 Vesicles perform a variety of functions because it is separated from

the cytosol.

 Vesicles are involve in metabolism, transport, buoyancy control, as

well as temporary storage of food and enzymes.

 They can also act as chemical reaction chambers. 3

 TYPES OF VESICLES

1. Natural Vesicles: formed during the process of secretion

(exocytosis), upake (endocytosis), and transport of materials within
the cytoplasm.

2. Artificial Vesicles: Artificially prepared vesicles called liposomes.

 They may be made up of only one phospholipid bilayer called
unilamellar liposome vesicles otherwise called multilamellar
liposome vesicles.
 Liposomes are being used as a medium to deliver drugs through the
skin.

4
 VESICLES FORMATION
 Vesicle formation provides a means of cellular entry for extracellular
substances and for recycling of membrane constituents.

 Vesicles are form based on three processes:

I. Fluid endocytosis
II. Phagocytosis
III. Receptor-mediated endocytosis

 Coat proteins induce curvature in membranes to form vesicles.

 Clathrin and caveolin are coat proteins that are well known to form
vesicles.
5
Fluid endocytosis (Adapted from Vander’s Human physiology: The mechanisms of
body function, 2014) 6
Phagocytosis (Adapted from Vander’s Human physiology: The mechanisms of body
function, 2014) 7
 SYNAPTIC VESICLES
 Synaptic vesicles are small membrane sacs that carry
neurotransmitters from the soma where they are produced to the
terminal button or pre-synaptic membrane where they are release.

 Small vesicles are produced by the Golgi apparatus located in the

soma and transported down the axon via current follow in the
cytoplasm.

 The released neurotransmitter molecules then bind to their cognate

receptors on the postsynaptic neuron, eliciting an array of chemical
and electrical changes.

8
Receptor-mediated endocytosis (Adapted from Vander’s Human physiology: The
mechanisms of body function, 2014)
9
 Synaptic
Vesicle

Synaptic Vesicles (Adapted from Vander’s Human physiology: The mechanisms

of body function, 2014)
10
 FORMATION OF SYNAPTIC VESICLES
 Delivery of synaptic vesicle components to plasma membrane.

 Endocytosis of synaptic vesicle components to form new

synaptic vesicles directly.

 Endocytosis of synaptic vesicle components and delivery to

endosome.

 Budding of synaptic vesicle from endosome.

 Loading of neurotransmitter into synaptic vesicle.

 Secretion of neurotransmitter by exocytosis in response to

action potential
11
Formation of Synaptic Vesicles (Adapted from Molecular Biology of
the Cell, 2002) 12
 SUMMARY
 A vesicle is a structure within or outside a cell, consisting of
liquid or cytoplasm enclosed by a lipid bilayer membrane.

 Vesicles may be natural or artificial.

 The synaptic vesicles are formed by two pathways, direct

and indirect pathways.

7/19/2019 13
 VESICULAR TRAFFICKING
INTRODUCTION

 Vesicular trafficking is at the basis of cellular life, it governs

cell communication via secretion and uptake of signalling
molecules (Anitel and Haflack 2011).

 Vesicles bud from the membrane of a donor compartment

and are subsequently transported to a different destination
compartment, where membrane fusion occurs.

14
 VESICULAR TRAFFICKING
INTRODUCTION
 Membrane trafficking can be divided into two basic
pathways based on the direction of travel, Exocytosis and
Endocytosis (McMahon, and Boucrot 2011).

 Exocytosis: this refers to the movement of cargo to the

plasma membrane or out of the cell.

 Endocytosis: this is the movement of cargo into the cell

from the plasma membrane

15
 TRANSPORT VESICLES

 Transport vesicles differ from one another in the type of cargo they
ferry from one site to another, the route they take (Faini et al., 2013).

 The presence or absence of proteins on the cytosolic surface which

can form a coat

 These transport vesicles bud off from one membrane and can
dynamically fuse with other membranes, or split up into smaller
vesicles by fission (Faini et al., 2013).

16
Schematic presentation of various membrane trafficking pathways. (Defining Mechanobilogy
2018). 17
The cellular mechanism that maintain neural polarity. (Marvin and Gary, 2016)
18
 TRANSPORT VESICLES

 The cytoskeleton is responsible for moving vesicles throughout the

cell.

 Most vesicles traffic along microtubules using kinesin or dynein

motors, although they can also use myosin II and Myosin V motors to
move along the actin network (Hirokawa et al., 2009).

19
 VESICULAR BUDDING AND FUSION

20
Steps of vesicular budding and fusion (Juan and Benjamin 2004).
 TRANSPORT VESICLES

 Members of the Rab small GTPase family are primarily responsible

for providing specificity.

 Rab proteins can be expressed on both transport vesicles and target

membranes, providing a further level of regulation (McMahoon and

Boucrot, 2015).

21
 TRANSPORT VESICLES

 SNARE proteins dock the transport vesicle at the correct membrane

location and catalyse membrane fusion.

 SNAREs bring the apposing membranes of the transport vesicle and

the target region closely together so that the lipids from the different
bilayers can mix, and this eventually results in fusion, and the release
of cargo (McMahoon and Boucrot, 2015).

22
Structure and Functions of Snares (Juan and Benjamin 2004) 23
 VESCULAR DOCKING
INTRODUCTION
 Docking is a process by which vesicles are juxtaposed to the plasma
membrane at the active zone which undergo exocytosis with the
presynaptic terminal in response to Calcium influx (Ting and Scheller,
2016).

 Docking also the association of secretory vesicles with the plasma

membrane, which precedes formation of the SNARE complex (Heidi
et al., 2009; Wang et al., 2016).

24
 ACTIVE ZONE

 In a nerve terminal, synaptic vesicle docking and release are

restricted to an active zone (Wang et al., 2016).

 The active zone is a highly organized protein scaffold that

docks synaptic vesicles close to release machinery and
presynaptic Ca2+ channels (Wang et al., 2016).

25
 VESICULAR DOCKING PROTEINS
 SNARE Proteins

 Munc 18-1

 Synaptotagmin-1

 Rab GTpase

 Rab effector

26
 VESICULAR DOCKING PROTEINS

 The SNARE protein complexes are the core protein machinery

involved in synaptic vesicle docking and fusion (Fortoul et al., 2016).

 The SNARE proteins form link between the vesicles and plasma
membrane providing a mechanism for zippering the two together
(Zilly et al., 2006; Fortoul et al., 2016).

27
 VESICULAR DOCKING PROTEINS

 The SNARE Protein complex applies an attractive force to overcome

the long range repulsion between the vesicles and the membrane
(Zilly et al., 2006; Fortoul et al., 2016).

 The balance between the attractive and repulsive forces lead to

formation of an equilibrium docked state,

 Even a single SNARE complex is able to bring a typical synaptic

vesicle to within a distance of -3nm from the membrane (Fortoul et
al., 2016).
 Other proteins such as Munc18, Munc13 and RIM proteins are
essential for the maintenance of a docked pool of synaptic vesicles
(Verhage and Sorensen, 2008;Wentzel et al., 2013).
28
 MECHANISMS OF VESICULAR DOCKING

 The transmembrane vesicle associated protein(VAMP) synaptobrevin

(v-SNARE) forms a four-helix bundle with the proteins SNAP-25,The
transmembrane protein syntaxin (t-SNARE), which are attached to the
neuronal plasma membrane,

 SNAP-25 contributes two helices (SN1 and SN2) to the bundle, while
both Syx and Syb contribute one helix each (Fortoul et al., 2016).

29
 MECHANISMS OF VESICULAR DOCKING

 After docking, priming occurs, which finally leads to vesicle-to

membrane fusion in the presence of Calcium (Fortoul et al., 2016).

 After fusion NSF interacts with the post-fusion SNARE complex on

the target membrane and dissociate them into individual SNAREs
(Jani and Setty, 2014).

 These SNAREs are removed for another fusion process (Jani and
Setty, 2014).

30
 Mechanisms

Vesicular docking mechanism (Deleo, 2003).

31
 Mechanisms

Vesicular docking mechanism (Deleo, 2003).

32
 Mechanisms

Exocytotic pathway from Docking to Fusion (Jani and Setty, 2014).

33
 APPLIED PHYSIOLOGY
 The SNAREs of the synaptic vesicle and presynaptic membrane
are the targets of two of the most potent bacterial toxins; those
responsible for botulism and tetanus (Deleo, 2003).

 These deadly toxins act as proteases whose only known

substrates are SNAREs (Deleo, 2003).

 Cleavage of the neuronal SNAREs by the toxins blocks the

release of neurotransmitters, which causes paralysis (Deleo,
2003).

34
 SUMMARY

 The process of synaptic vesicle docking and fusion can be viewed as

deformation of mechanical system, in which a synaptic vesicle, a
nearly spherical lipid bilayer shell, is brought in proximity to the
plasma membrane, a nearly flat lipid bilayer, under the influence of
an attractive forces exerted by SNARE complex.

35
 VESICULAR PRIMING
INTRODUCTION

 The release of neurotransmitter from synaptic vesicles

represents the final event by which pre synaptic neurons send
their chemical signal to the receiving post synaptic neurons.

 Prior to fusion, synaptic vesicles undergo a series of

maturation events, most notably the membrane-delimited
docking and priming steps. (Becherer and Rettig, 2006)

36
 WHAT IS PRIMING IN SYNAPTIC IMPULSE
TRANSMISSION?
 Priming is a process that converts synaptic vesicles to a state
of competence for calcium triggered fusion with the active
zone membrane

 Priming is a process that makes the vesicle release competence

such that it can fuse with the plasma membrane when the
trigger (calcium) arrives (Matthijs and Jakob, 2008).

 Once synaptic vesicles have docked at the active zone, they

must be transformed into releasable vesicles. This process is
called PRIMING.

 It is a calcium dependent step and it typically but not always

occurs after docking (Jahn and Fasshauer, 2012). 37
 ROLE OF PRIMING

 Prepares the synaptic vesicles for fusion in response to

calcium influx

 Helps in bringing synaptic vesicle membrane and plasma

membranes into close proximity with the aid of SNARE-
containing complexes and thereby facilitating membrane
fusion.

 Priming also determines the probability of the vesicle

membrane fusing with the presynaptic membrane when a
nerve impulse arrives

38
 REQUIREMENTS FOR PRIMING IN SYNAPTIC
VESSICLES

 Two proteins, Munc-13 and CAPS (Calcium-dependent activator

protein for secretion), have been shown to be crucial for vesicle
priming.
 These proteins are important for the initiation of SNARE
(Soluble N-ethylmaleimide Attachment protein Receptor)
complex formation and help to stabilize synaptic vesicles in a
fusion competent state (Sudhof, 2004, Jockusch et al., 2007).
 Munc13 catalyzes opening of Syntaxin1 to initiate SNARE
assembly through the minimal activity domain of Munc13
(MUN) domain (Basu et al., 2005; Ma et al., 2011; Wang et al.,
2017)
 CAPS facilitates SNARE complex formation in a manner
dependent on interaction with individual SNAREs and/or the
partially assembled SNARE complex (James et al., 2009; Daily
et al.,2010; Khodthong et al., 2011)

39
 40
Neurocellular priming (Zhou et al.,2019)
Munc-13 function in priming (Current opinion in Neurobiology, 2012) 41
 REGULATION OF SYNAPTIC VESICLE PRIMING

 Fusion of synaptic vesicles with the plasma membrane

requires the formation of the soluble N-ethlymaliemide-
sensitive factor attachment protein receptor (SNARE)
complex.

 Munc 13 proteins regulate this SNARE complex assembly

process and thereby prime synaptic vesicles to fusion
competence

 Munc 13-mediated synaptic vesicle priming is absolutely

essential for synaptic vesicle fusion.

42
 EXOCYTOSIS
INTRODUCTION

 Exocytosis is defined as the transport and fusion of secretory

vesicles with the plasma membrane and the extracellular
space.(luzio et al.,2007).

 It is a form of active transport and bulk transport in which a

cell transport molecules (e.g neurotransmitters and proteins)
out of the cell, by secreting them through an energy-
dependent process.
 BASIC PROCESS OF EXOCYTOSIS

 Vesicles containing molecules are transported from within the

cell to the cell membrane.

 The vesicle membrane attaches to the cell membrane.

 Fusion of the vesicle membrane with the cell membrane

releases the vesicle contents outside the cell.
 FUNCTIONS OF EXOCYTOSIS

 It allows cells to secrete waste substances and molecules, such

as hormones and proteins.

 Exocytosis is also important for chemical signal messaging

and cell to cell communication.

 exocytosis is used to rebuild the cell membrane by

fusing lipids and proteins removed through endocytosis back
into the membrane.
 TYPES OF EXOCYTOSIS

 Regulated exocytosis ( Ca2+triggered non constitutive

exocytosis)

 Non Regulated exocytosis (Non Ca2+triggered constitutive).

 REGULATED EXOCYTOSIS

 It relies on the presence of extracellular signals for the

expulsion of materials within vesicles.

 occurs commonly in secretory cells and not in all cell types.

 Ca2+ triggered non-constitutive exocytosis requires an external

signal, a specific sorting signal on the vesicles, a clathrin coat,
as well as an increase in intracellular calcium.
 NON REGULATED EXOCYTOSIS

 This type of exocytosis is non calcium triggered.

 Constitutive exocytosis functions to deliver membrane

proteins and lipids to the cell's surface and to expel substances
to the cell's exterior.
PROCESSES INVOLVED IN EXOCYTOSIS
 STEPS INVOLVED IN EXOCYTOSIS

 Trafficking: it involves the transportion of vesicles to the cell

membrane.Is powered by the motor proteins: kinesins,
dyneins, and myosin.

 Tethering: When the vesicles reach the cell membrane, they

become linked to and pulled into contact with the cell
membrane
 Docking: Docking involves the attachment of the vesicle
membrane with the cell membrane. The phospholipids bilayers
of the vesicle membrane and cell membrane begin to merge.

 Priming: occurs in regulated exocytosis and not in

constitutive exocytosis.
 Fusion: There are two types of fusion that can take place in
exocytosis, which are:

 Complete fusion

 Kiss and run fusion

 COMPLETE FUSION

• In complete fusion, the vesicle membrane fully fuses with the

cell membrane. The energy required to separate and fuse the
lipid membranes comes from ATP. The fusion of the
membranes creates a fusion pore, which allows the contents of
the vesicle to be expelled as the vesicle becomes part of the
cell membrane.
 KISS AND RUN

 In kiss and run fusion, the vesicle temporarily fuses with the
cell membrane long enough to create a fusion pore and release
its contents to the exterior of the cell. The vesicle then pulls
away from the cell membrane and reforms before returning to
the interior of the cell.
 VESICLE RETRIEVAL

 Retrieval of synaptic vesicles occurs by endocytosis. Some

synaptic vesicles are recycled without a full fusion into the
membrane (kiss-and-run fusion), while others require a
complete reformation of synaptic vesicles from the membrane
by a specialized complex of proteins.
 EXOCYTOSIS IN NEURON

 Some neurons communicates through the transmission of

neurotransmitters.

 A synaptic vesicle filed with neurotransmitters In the presynaptic

neuron fusses with the presynaptic membrane releasing
neurotransmitters into the synaptic cleft (Gap between neurons).

 The neurotransmitters can then bind to receptors on the post

synaptic neurons.
 EXOCYTOSIS IN NEURONS

 Synaptic vesicle exocytosis occurs in neurons of the nervous

system.

 Nerve cells communicate by electrical or chemical

(neurotransmitters) signals that are passed from one neuron to
the next.
 EXOCYTOSIS IN NEURONS

 Neurotransmitters are transmitted by exocytosis.

 They are chemical messages that are transported from nerve to

nerve by synaptic vesicles.

 Synaptic vesicles are membranous sacs formed by endocytosis

of the plasma membrane at pre-synaptic nerve terminals.
 EXOCYTOSIS IN NEURONS

 Once formed, these vesicles are filled with neurotransmitters

and sent toward an area of the plasma membrane called the
active zone.

 The synaptic vesicle awaits a signal, an influx of calcium ions

brought on by an action potential, which allows the vesicle to
dock at the pre-synaptic membrane.

 Actual fusion of the vesicle with the pre-synaptic membrane

does not occur until a second influx of calcium ions occurs.
 EXOCYTOSIS IN NEURON

 After receiving the second signal, the synaptic vesicle fuses

with the pre-synaptic membrane creating a fusion pore.

 This pore expands as the two membranes become one and the
neurotransmitters are released into the synaptic cleft (gap
between the pre-synaptic and post-synaptic neurons). The
neurotransmitters bind to receptors on the post-synaptic
neuron.

 The post-synaptic neuron may either be excited or inhibited by

the binding of the neurotransmitters.
 SUMMARY

 During exocytosis, cells transport substances from the interior

of the cell to the exterior of the cell.

 This process is important for the removal of waste, for

chemical messaging between cells, and for rebuilding the cell
membrane.

 Exocytotic vesicles are formed by the Golgi apparatus,

endosomes, and pre-synaptic neurons.
 SUMMARY

 Three pathways of exocytosis are constitutive exocytosis,

regulated exocytosis, and lysosome mediated exocytosis.

 Steps of exocytosis include vesicle trafficking, tethering,

docking, priming, and fusing.

 Vesicle fusion with the cell membrane may be complete or

temporary.

 Exocytosis occurs in many cells including pancreatic cells and

neurons.
 DIFFERENCES BETWEEN NEUROTRANSMITTERS AND
HORMONES
INTRODUCTION

 Neurotransmitters and Hormones are two different types of

chemicals that signal from one part of the body to the other.

 Both chemicals play an important part in the body’s

physiology.

 Hormones can be either proteins, lipids or cholesterol-based

molecules while, Neurotransmitters are proteins (Wiley, 2015)

63
 NEUROTRANSMITTERS

 Neurotransmitters are endogenous chemical substances that act as

a mediator for the transmission of nerve impulse from one neuron
across a chemical synapse, such as a neuromuscular junction,
another neuron, muscle cell, or gland cell (Lodish etal 2000).

 Their exact numbers are unknown, but more than 200 chemical
messengers have been uniquely Identified (Cherry and Kendra,
2014).

64
 SOME NEUROTRANSMITTERS

Excitatory Inhibitory Neurotransmitters With

Neurotransmitters Neurotransmitters Excitatory And Inhibitory
Function
1. Acetylcholine 1. Gamma-aminobutyric acid 1. Noradrenaline
2. Nitric oxide 2. Glycine 2. Adrenaline
3. Histamine 3. Serotonin 3. Dopamine
4. Glutamate
5. Aspartate

65
Neurotransmission mechanism (Source: Biology learnspot.com)
 MAIN CRITERIA FOR IDENTIFYING
NEUROTRANSMITTERS

 The chemical must be synthesized in the neuron or otherwise

be present in it.

 When the neuron is active, the chemical must be released and

the release is calcium dependent to produce a response in some
target.

67
 MAIN CRITERIALS FOR IDENTIFYING
NEUROTRANSMITTERS

 The same response must be obtained when the chemical is

experimentally placed on the target.

 A specific receptor for the chemical must be present on the

target and a mechanism must exist for removing the chemical
from its site of activation. (Breedlove and Watson, 2013).

68
 HORMONES

 Hormones are a class of signalling molecules produced by

endocrine glands in multicellular organisms that are
transported by the circulatory system to target distant
organs to regulate physiology and behaviour (Neave, 2008).

 Hormones have diverse chemical structures, mainly of three

classes:
 Eicosanoids,
 Steroids, and;
 Amino acid/protein derivatives (amines, peptides, and proteins).
(Neave, 2008)

69
 DIFFERENCES BETWEEN HORMONES
AND NEUROTRANSMITTERS

S/No NEUROTRANSMITTER HORMONE

1. OCCURENCE Neurotransmitters are found Hormones are found both
only in animals in plants and animals
2. ORGAN SYSTEM Neurotransmitters belong to Hormones belong to the
the nervous system endocrine system
3. CHEMISTRY Neurotransmitters are Hormones are protein
protein based molecules based molecules,
(Amino acids, peptides and eicosanoids and steroids.
monoamines) and
gasotransmitters
4. POINT OF Neurotransmitters are Hormones are produced
RELEASE released by presynaptic in the endocrine glands
nerve terminal into the and are secreted in to the
Synapse blood stream

70
S/N NEUROTRANSMITTER HORMONE

5. TRANSMISSION Neurotransmitters are transmitted Hormones are transmitted

across the synaptic cleft through the blood. Act on
receptor cells throughout the
body via the circulatory
system
6. POINT OF ACTION Neurotransmitters are in direct Hormones act on a distant
apposition to their target cells. site from where it is
(Target cells of neurotransmitters produced
can be specific neurons or other
cells)
7. RESPONSE Neurotransmitters quickly make Hormones take few minutes
the response, usually within to few days to make their
milliseconds response
8. FUNCTION Neurotransmitters are involved in Hormones have diverse roles
Neurotransmission (transmission and functions in controlling
of nerve signals). physiological , psychological
patterns and behaviour
71
S/N NEUROTRANSMITTER HORMONE

9. ROLE Neurotransmitters only Hormones are capable of

stimulate the postsynaptic
neurons
10. EXAMPLES Examples include serotonin,
dopamine, norepinephrine,
epinephrine, glutamate,
aspartate, glycine, nitrogen
oxide, and carbon monoxide
11. SIGNAL RANGE Neural signals are restricted to
pre-existing nerve tracts
12. NATURE OF Neural signalling is an all-or-
ACTION nothing (digital) action.
regulating target organs or
tissues
Examples include oxytocin,
cortisol, testosterone, and
estrogen (in animals) and
abscisic acid, cytokines, and
gibberellins (in plants)
Hormonal signals can travel
virtually anywhere in the
circulatory system.
Hormonal signalling is an
action that can be
continuously variable as
dependent upon hormone
concentration

72
S/N NEUROTRANSMITTER HORMONE

13. ESTABLISHED The exact numbers of List of known human

QUANTITY neurotransmitters are hormones has it at about
unknown but more than 200 50 or so.
of them have been uniquely
identified

14. FREQUENCY OF Can relay the same signal Once a hormone binds
TRANSMISSION multiple times with its receptor, it cannot
impart its signal a second
time

15. QUALITY OF Voluntary or involuntary Involuntary action

RESPONSE action
ACTION

73
 NEUROTRANSMITTERS RECEPTORS
INTRODUCTION

 The sensory receptors are specialized structures located at the

peripheral ends of sensory neurons, and they may be a part of
the neuron or a separate organ.

 The act as detectors and transducers.

 They inform the central nervous system about the changes

occurring inside and outside the body.

2
 PROPERTIES OF RECEPTORS

 Specificity: ability to respond to a specific sensation.

 Excitability(Receptor Potential):This is the property of
responding to stimuli by generating action potentials
 Discharge of impulses: It occurs according to the weber
Fechner law which states that “the frequency of discharge
from receptors is directly proportional to the logarithm of
intensity of the applied stimulus”.
 Adaptation: this is a decline in the frequency of discharge of
action potential due to maintained stimulation by stimuli of a
constant strength.

3
 CLASSIFICATION OF RECEPTORS

Receptors are classified based on: Specificity, Location and Adaptation

Receptors

Specificity Location Adaptation

Chemo Exteroceptor Interoceptor Slow

Mechano Rapid
Receptor Receptor
Thermo Nociceptor Moderate
Receptor

4
 NEUROTRANSMITTER RECEPTORS

 Neurons in the human brain communicate with one another by

releasing chemical messengers called neurotransmitters.
 Neurotransmitters evoke postsynaptic electrical responses by
binding to members of a diverse group of proteins called
neurotransmitter receptors.
 There are two major classes of neurotransmitter receptors
1. Ionotropic receptors
2. Metabotropic receptors

5
 IONOTROPIC RECEPTOR

 These receptors are linked directly to ion channels. They

contain two functional domains: an extracellular site and a
membrane-spanning domain.
 Thus ionotropic receptors combine transmitter-binding and
channel functions into a single molecular entity (they are
also called ligand-gated ion channels).
 In neuromuscular junction, transmission of impulses
involve the release of acetylcholine from presynaptic
membrane that move through the synaptic cleft to
postsynaptic membrane.
 Then, the acetylcholine molecules bind with its
receptor(nicotinic receptor) causing opening of sodium
channels in the postsynaptic membrane.
6
Mechanism of ionotropic receptors.
Retrived from https://gozasso.com/kikkenlab/en/036-ligand-gated-ion-channel/ 7
 METABOTROPIC RECEPTORS

 These receptors are called metabotropic receptors, because

the eventual movement of ions through a channel depends
on one or more metabolic steps.
 These receptors do not have ion channels as part of their
structure; instead, they affect channels by the activation of
intermediate molecules called G-proteins, hence they are
also called G-protein-coupled receptors.
 Neurotransmitter binding to metabotropic receptors
activates G-proteins, which then dissociate from the
receptor and interact directly with ion channels or bind to
other effector proteins, such as enzymes, that make
intracellular messengers that open or close ion channels.

8
 G- PROTEIN

 G proteins or guanosine nucleotide-binding proteins are the

membrane proteins situated on the inner surface of cell
membrane. These proteins play an important role in the
formation of cAMP.
 Each G protein molecule is made up of trimeric (three)
subunits called α, β and γ subunits.
 The α-subunit is responsible for most of the biological actions.
It is bound with guanosine diphosphate (GDP) and forms α-
GDP unit.
 The α-subunit is also having the intrinsic enzyme activity
called GTPase activity.
 The β and γ subunits always bind together to form the β-γ
dimmer. 9
SEQUENCE OF EVENTS IN THE FORMATION OF
CAMP
 Hormone binds with the receptor in the cell membrane and forms the hormone-
receptor complex
 It activates the G protein
 G protein releases GDP from α-GDP unit
 The α-subunit now binds with a new molecule of GTP, i.e. the GDP is
exchanged for GTP
 This exchange triggers the dissociation of α-GTP unit and β-γ dimmer from the
receptor
 Both α-GTP unit and β-γ dimmer now activate the second messenger pathways
 The α-GTP unit activates the enzyme adenyl cyclase, which is also present in
the cell membrane.
 Most of the adenyl cyclase protrudes into the cytoplasm of the cell from inner
surface of the cell membrane
 Activated adenyl cyclase converts the adenosine triphosphate of the cytoplasm
into cyclic adenosine monophosphate (cAMP)
 Cyclic AMP executes the actions of hormone inside the cell by stimulating the
enzymes like protein kinase A. Cyclic AMP produces the response, depending
upon the function of the target cells through these enzymes.
10
Mechanism of metabotropic receptor.
Essential of Medical Physiology by K Sembulingam and Prema Sembulingam 6 th 11
Edition.
NEUROTRANSMITTERS, THEIR RECEPTORS AND LOCATION.

S/N NEUROTRANSMITTER RECEPTOR CLASS LOCATION

IONOTROPIC METABOTROPIC
1 Acetylcholine Nicotinic Skeletal muscle
Muscarinic Heart, smooth muscle
2 GABA GABAA Limbic system
GABAB CNS and autonomic
division of the PNS
3 Dopamine D1 and D2 like Cortex, striatum and
limbic system, CVS
4 Epinephrine and α –adrenergic Blood vessels, heart,
Norepinephrine β -adrenergic bronchioles
5 Serotonin 5HT3 5HT Enterochromaffin
cells in the gut, CNS
6 Glutamate NMDA, AMPA,kainate Metabotropic CNS
receptor glutamate receptor

KEYS: 5HT: 5-hydroxytryptamine receptor

NMDA: N-methyl-D-aspartate
AMPA: α-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate
PNS: Peripheral nervous system
CNS: Central nervous system
CVS: cardiovascular system
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 POST SYNAPTIC POTENTIALS
INTRODUCTION
 Postsynaptic potential is the potential difference between the
intracellular fluid and extracellular fluid across the membrane of the
postsynaptic cell.

 A synapse is structure that permit neuron ( nerve cell) pass electrical
current, chemical substance or information to another neuron or
target
 organ.(Sudhof, 2018).

Types of synaptic potential

 Electrical
 chemical

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 POSTSYNAPTIC POTENTIALS

 Electrical synapses utilize gap junctions which allow flow of

current between pre- and postsynaptic cells. (Purves, D. I., 2004)

 Chemical synapses are cell to cell communication via

use of neurotransmitters.

TYPES OF POSTSYNAPTIC POTENTIAL

 Excitatory post synaptic potential (EPSP)

 Inhibitory post synaptic potential (IPSP)

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 Excitatory post synaptic potential (EPSP) – resulting from
depolarization of the postsynaptic membrane. (Jones et al., 2017).

 Inhibitory post synaptic potential (IPSP) – resulting from the

hyperpolarization of the postsynaptic membrane(Jones et al., 2017).

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(Purves et all., 2004) 88
CAUSES OF CHANGE IN POSTSYNAPTIC
POTENTIAL

(Sembulingam and senbulingam 2012)89

INHIBITORY POSTSYNAPTIC POTENTIAL (IPSP)

 Activation of the receptors that open potassium channel – allows

more potassium ion to leave, making the cell more negative inside

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91
EPSP IPSP

Is cause by influx of positive ions Is cause by influx of negative ions

it makes the post synaptic membrane excited it takes post synaptic membrane less excited

it bring the post synaptic membrane close to It takes the post synaptic membrane away from
threshold the threshold

(Wikimedia foundation., 2019)

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 SIMILARITIES
 Both are types of post synaptic potential

 Both occur at post synaptic membrane

 Both are mediated by ligand gated ion channels which are open by
the binding of a neurotransmitter. (Wikimedia foundation 2018).

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94
 FATE OF NEUROTRANSMITTERS AFTER
NEUROTRANSMISSION
INTRODUCTION

 After synaptic transmission neurotransmitters in the synaptic

cleft must be cleared through the following processes:

 Simple diffusion

 Enzymatic destruction

 Reuptake (Harris, 2018).

Fate of Neurotransmitters after

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Neurotransmission
 FATE OF NEUROTRANSMITTERS AFTER
NEUROTRANMISSION

 Neurotransmitter can simply diffuse out of the synaptic cleft

away from the receptors, into nearby blood vessels.

 Neurotransmitter can be destroyed directly using Monoamine

Oxidases (MAO) or CatecholOMethyltransferase (COMT)
(Marco, et al., 2008).

 MAO catalyzes the oxidative deamination of monoamine

neurotransmitters.

 COMT is responsible for degrading catecholamines (Michael and

Matthew, 2018).
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Oxidative deamination of monoamine neurotransmitters (adapted from Marco et al., 2008).

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 RE-UPTAKE

 These is the reabsorption of neurotransmitters by neurotransmitters

transporters (Sembulingam and Sembulingam, 2012).

 Some neurotransmitters are too large and hydrophilic hence,

specific transport proteins are necessary for the reabsorption of
neurotransmitters.

 Re-uptake can either be through the pre-synaptic cell membrane or

glial cells membrane(Südhof, 2018).

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 REUPTAKE

 Reuptake is necessary for normal synaptic physiology

because;

 It regulates the level of neurotransmitter.

 It allows for the recycling of neurotransmitters (David et

al., 2008).

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 NEUROTRANSMITTER TRANSPORTERS

 These are ion-coupled carriers which regulate the accumulation of

the neurotransmitter (Rudnick, 1998).

 Reuptake transporters are members of a large super-family of

neurotransmitter which include:

 Na and Cl dependent symporters.

 Na and K dependent antiporters.

Fate of Neurotransmitters after

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Neurotransmission
 MECHANISM OF ACTION

 Na and Cl dependent Symporters: This transport protein is

Na+ and Cl− ion dependent and uses transmembrane ion
gradients and electrical potential (Claxton et al., 2010).

 A substrate could be released only to the side from which it

bound.

 But the transporter has the capacity to convert the binding site
so that it is accessible from the other side of the membrane.

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Ion Coupling Stoichiometry for Plasma membrane Neurotransmitter Reuptake

(Adapted from Rudnick, 1998). 102

Mechanism for Ion-Coupled Neurotransmitter Transport (Adapted from

Rudnick, 1998). 103

 Na and K-Dependent Antiporters

 The Na- and K-dependent anti-porters are indirectly

driven by the Na+/K+ATPase which generates
gradients of Na+ (out to in) and K+ (in to out)

 This process creates a membrane potential (inside

negative) (Rudnick 1998).

Fate of Neurotransmitters after

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Neurotransmission
Na- and K-dependent Antiporters (Adapted from Rudnick, 1998).

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 Na and K-Dependent Antiporters

 Cytoplasmic ATP acts at the plasma membrane Na+/K+-

ATPase (white circle)

 This pump Na+ out of the cell and K+ into the cell. Since 3
Na+ ions are pumped per 2 K+ ions, a membrane potential
(interior negative) is generated.

 Cl- ion is driven out of the cell by interior negative

membrane potential, generating a Cl- gradient (Rudnick
1998).

Fate of Neurotransmitters after

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Neurotransmission
Specialized Neurotransmitter Transporters in Astrocytes (Adapted from Yongjie and Jeffrey

2008). 107
 SUMMARY
 After synaptic transmission, neurotransmitters must be cleared through
at least one of the following processes:

 Simple diffusion; enzymatic destruction; and reuptake.

 Reuptake is the reabsorption of neurotransmitters by neurotransmitters

transporters located at the plasma membrane of an axon terminal or
glial cells after it has perform its function.

 Reuptake transport protein uses transmembrane ion gradients and

electrical potential to transport neurotransmitter

 Neurotransmitter Sodium Symport (NSS) transporters take advantage of

both Na+ and Cl− gradients, inwardly directed across the membrane.

 The Na- and K-dependent anti-porters are indirectly driven by the

Na+/K+ATPase which generates gradients of Na+ (out to in) and K+ (in
to out) and in the process creates a membrane potential (inside
negative).
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 THANK YOU

FOR

YOUR AUDIENCE
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