INTRODUCTION TO AUTONOMIC PHARMACOLOGY: Mechanism of neurotransmission at in the autonomic nervous system

PHC 452

Peripheral Nervous System

Controls skeletal muscle Controls smooth & cardiac muscle & glands Autonomic Nervous System

Somatic Nervous System

One Neuron Efferent Limb

Two Neuron Efferent Limb

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Peripheral Nervous System Somatic Nervous System Autonomic Nervous System

Parasympathetic Sympathetic Nervous System Nervous System Selective Activation Skeletal Muscle Diffuse Activation

Glands, Smooth Muscle & Cardiac Muscle

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Objectives
To know: The autonomic nervous system neurotransmitters. Neurotransmitter functions:
synthesis storage release receptor activation termination of activity

NE biosynthesis and metabolism ACh biosynthesis and metabolism Receptors of the autonomic nervous system
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Biochemical Aspects of ANS

Neurotransmission: Nerve impulses are transmitted from one nerve to another or to a tissue by chemicals (neurotransmitters). ANS Neurotransmitters:
Acetylcholine Norepinephrine Dopamine
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Summary of neurohumoral transmission:

Norepinephrine: NT at most postganglionic fibers of the SNS & is released by adrenal medulla chromaffin cells into the circulation

Sympathetic cholinergic nerve

Acetylcholine
Released by:

all preganglionic fibers of the SNS and the PSNS preganglionic nerves to the adrenal medulla all postganglionic nerves of the PSNS some postganglionic nerves of the SNS (sweat glands)

sympathetic cholinergic nerves

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The adrenal medulla

The adrenal medulla belongs to the sympathetic division, since all preganglionic fibers that innervate its cells originate in the thoracolumbar region of the spinal cord. Its cells are homologous to postganglionic neurons Preganglionic nerves terminate on modified nerve cells which release adrenaline and noradrenaline into the bloodstream. These circulating catecholamines are removed from the blood slowly and maintain neuronal sympathetic activity.

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Adrenal Medulla
Cells of the adrenal medulla secrete epinephrine (EPI; adrenaline): a hormone structurally and functionally similar to NE as well as small amounts of NE itself ratio of EPI : NE is 4:1 (80% EPI & 20% NE)

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Maintain neuronal sympathetic activity, also responsible for sustained effect

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Key features of neurotransmitter function:

Synthesis Storage Release of NT by the nerve terminal Activation of receptor: Interaction of NT with the postjunctional cell and initiation of postjunctional activity 5. Termination of neurotransmitter activity 1. 2. 3. 4.

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Uptake of precursor

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Mechanism of NT release
Action potential at axon terminal depolarisation of membrane opening of voltage-gated calcium channels. a high, but transient increase in intracellular Ca2+ near NT storage vesicles activation of calmodulin (Ca2+-binding protein) activation of Ca2+/calmodulin protein kinase catalyses phosphorylation of proteins associated with storage vesicles (including synapsin I). Synapsin I binds to actin on cytoskeleton and interacts with other proteins (synaptobrevin, synaptophysin, synaptoporin) to initiate docking & fusion of storage vesicles with the cell membrane Exocytosis of NT
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General Features of Peripheral Autonomic Neurotransmission

NT = Neurotransmitter

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Autonomic Neurotransmission

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General Features of Peripheral Autonomic Neurotransmission

Nerve Impulse Membrane Depolarization of Pre- or Postganglionic Fiber Calcium Entry into Varicosity Exocytosis of NT Activation of NT Receptors Depolarization of Postganglionic Fiber or Response of Effector Cell

Diffusion of NT Across Neuroeffector Junction or Synapse

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Neurohumoral deactivation
3 potential mechanisms: 1.diffusion of NT away from the junction 2.enzymatic destruction of NT 3.reuptake of NT by the prejunctional fiber The relative importance of each mechanism at a given junction depends on the type of transmission that occurs at that junction.
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Neurohumoral degradation
The sympathetic division tends to exert its effects for sustained periods of time, while the parasympathetic division produces responses of short duration. Sustained sympathetic effects are due in part to circulating EPI and NE from the adrenal medulla but also due to differences in neurotransmitter deactivation mechanisms. At most sympathetic neuroeffector junctions, the major deactivation mechanism for NE is via reuptake with diffusion playing a secondary role. These processes occur slowly relative to deactivation of ACh through enzymatic degradation by AChE.
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Synthesis of ACh

CAT: choline acetyltransferase (present in axonal terminals) Acetyl CoA: synthesized in mitochondria (present in large numbers in axonal terminals) Choline: transported from the plasma (rate-limiting step that requires a Na+-dependent carrier)

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Storage of ACh
Ach is transported into synaptic vesicles via a proton antiporter (inhibited by vesamicol)

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Release of ACh
(You should be able to explain this now)

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Degradation of ACh
ACh is rapidly hydrolyzed by acetyl cholinesterase present in cholinergic synapses
There is very little leak of ACh from the synaptic cleft.

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Degradation of ACh Hydrolysis by acetylcholinesterase (AChE): present in cholinergic synapses.

AChE rapidly converts ACh to choline and acetic acid with a turnover time of 150 sec.

AChE is anchored to the plasma membrane through a glycolipid

Binding of substrate. Formation of a transient intermediate involving the -OH group of Serine 203 of AChE. Loss of choline and formation of acetylated enzyme. 61 Deacylation of enzyme by attack with H2O.

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ACh Synthesis
Rate limiting step: uptake of choline into nerve terminal

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SUMMARY OF CHOLINERGIC NEUROTRANSMISSSION

Neuroeffector Junction

Parasympathetic/Sympathetic Preganglionic Fiber or Parasympathetic Postganglionic Fiber

Parasympathetic or Sympathetic Postganglionic Fiber or Effector Cell

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NE synthesis

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Norepinephrine Biosynthesis
Ratelimiting step

Stored with chromogranin, dopamine hydroxylase, ATP

PNMT: phenylethanolamine-N-methyl transferase, DOPA: Dihydroxyphenyl alanine

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Summary of Catecholamine Biosynthesis

Tyrosine Rate-limiting step DOPA

Dopamine SYMPATHETIC NERVE Norepinephrine ADRENAL GLAND Epinephrine

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 2 pools of NE within neuron:

Mobile: includes newly synthesized NE, preferentially released during depolarisation Reserve

90-95% of NE stored in vesicles, 5-10% found in cytoplasm. Autonomic axon resembles a string of beads as it passes among the smooth muscle fibers in blood vessels, intestines ect. The beaded varicosities release NT near relatively few directly innervated effector cells.
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Degradation of catecholamines
Primary mechanism: reuptake (Reuptake 1) by the prejunctional fiber Diffusion of NE is secondary. The reuptake process is considerably more effective for NE than for EPI. In mammals, two enzymes are present that can metabolize NE and EPI:
monoamine oxidase (MAO): mitochondrial of adrenergic nerve terminals, found in widely distributed especially in liver, kidney, brain catechol-O-methyl transferase (COMT): a cytoplasmic, extraneuronal enzyme, widely 70 distributed.

Degradation of catecholamines
neither MAO nor COMT plays a significant role in the deactivation of NE and EPI at sympathetic neuroeffector junctions but, they are important for the metabolism of circulating EPI (and NE) from the adrenal medulla and for the metabolism of exogenously administered EPI and NE.

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Noradrenaline Metabolism

Active, requires ATP & Mg2+

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Synthesis and degradation of catecholamines

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Receptors of the ANS

Named after the nerve type that innervates them

Cholinergic Receptors
Nicotinic
NM NN

Adrenergic Receptors

1 2

1 2 3

Muscarinic
M1 5

The receptors on which Ach acts

The receptors on which NE and Epi act 1: predominant postjunctional membrane receptor 2: located both prejunctionally & postjunctionally 74

Multiple subtypes of adrenergic receptors Molecular cloning: 1-adrenergic receptors: 1A,, 1B,, 1D 2A,, 2B,, 2C 1, 2, 3

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Cholinergic Receptors:

Nicotinic receptors:
Located on all autonomic ganglia (PSNS and SNS) and on adrenal medulla (NN)
Activation causes transmission of nerve impulses at all PS and S ganglia and release of Epi and NE from adrenal medulla

Located on skeletal muscle at the neuromuscular junction (NM)

Activation causes muscle contraction

The 2 receptor subtypes are structurally different based on different responses to drugs
ganglionic blockers and neuromuscular blockers
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Cholinergic Receptors:

Muscarinic Receptors:
Located on PS postganglionic effector tissue most internal organs, including the CV system, the respiratory system, the gastrointestinal system and the genitourinary systems Activation by ACh leads to stimulation or inhibition of the effector tissue function
Activation of M-receptors in heart causes a in HR; activation of M-receptors in GI causes in GI activity

5 receptor subtypes

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Thank You

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