Intro to Cholinergic Pharmacology Flashcards
What are the 2 methods of ACh metabolism?
- De novo synthesis (minor)
2. Alternative synthesis (major) - most choline is recycled from ACh
Describe the steps of de novo ACh synthesis
- ACh synthesis is a 2-step conversion of serine to choline.
- Step 1 requires enzyme serine decarboxylase.
- Step 2 requires choline N-methyl transferase and S-adenosyl methionine. - The enzyme choline acetyltransferase (ChAT) is responsible for the acetylation of choline. ChAT resides in the cytoplasm so ACh must be in the cytoplasm. Acetyl-CoA comes from the inner layer of the mitochondria and comes out of the mitochondria as citrate.
What modifications must ACh undergo to have activity in the brain and nervous system?
ACh is the major NT in the brain, but choline is charged so there is a BBB to overcome. Once inside the brain, choline remains charged and has to be transported into the neurons into the brain. ChAT converts choline into ACh by adding an acetyl group which allows it to be an active NT in neurons.
How is ACh stored and released?
ACh is stored in synaptic vesicles. Cholinergic vesicles contain 2 important transporters which are required to concentrate ACh:
1. proton ATPase
2. ACh-H+ antiport
This has reverse vs. depot pools. ACh-H+ antiport concentrates ACh. The vesicle contains ATP and heparin proteoglycans. Every time ACh is released, heparin sulfate proteoglycans are also released. There are 2 types of pools for release:
1. readily release pool
2. reverse pool - this pool waits for an action potential to be released, then it moves into position for supply.
Release of ACh can be done either by spontaneous or stimulated release.
- spontaneous release from terminal has constitutive activity of the NT at the synapse and produces a synaptic potential
- stimulated release relies on stimulation by an action potential.
Describe the function and mechanism of SNARE proteins
SNARE proteins anchor and release vesicles that contain NTs. There are SNARE proteins on the vesicles (v-SNAREs): synaptotagmin and synaptobrevin as well as SNARE proteins on the terminal membrane: SNAP-25 and Syntaxin.
Synaptotagmin is a Ca2+ receptor and Synaptobrevin binds to syntaxin and SNAP-25 on the membrane to form the ternary complex.
There are 3 steps to vesicle release of NT: 1. docking, 2. priming, and 3. fusion.
n-sec-1 binds syntaxin in the initial state, but it disassociates in order for synaptobrevin to interact with syntaxin and SNAP-25.
This complex tightens and leads to fusion and exocytosis when Ca2+ enters the neuronal terminal and binds to synaptotagmin. This leads to release of NT into the synaptic cleft. Then ATP is required to disassemble the ternary complex by releasing synaptobrevin from syntaxin and SNAP-25 to disassociate from synaptobrevin.
How does the botulinum toxin drug act upon SNARE proteins?
Botulinum toxin degrades the SNAP-25 SNARE protein on the terminal membrane and thus prevents vesicle docking on the membrane. This prevents vesicle fusion and exocytosis of ACh, so that it is unable to be released from the vesicle into the synaptic cleft.
What are 3 ways that ACh is degraded and recycled and describe their mechanism of action.
Metabolism of ACh:
- ACh is made from choline and acetyl CoA
- ACh is broken down by AChase in synpatic cleft
- Choline is transported back to the axon terminal
- Acetylcholinesterase
- this is found in the vicinity of cholinergic synapses and in RBCs
- it has a high affinity for ACh and degrades the NT in the synaptic cleft - Pseudocholinesterases (plasma or butyrlcholinesterase_
- found in plasma
- T1/2 range from 8-16 hours
- concentrations may directly affect succinylcholine therapy - Fate of metabolites
- choline is taken up by special choline transporters into pre-synaptic neurons
- coenzyme A is released from the mitochondria
How is ACh activity implicated in pts with Alzheimer’s?
Acetylcholinesterase will break down ACh and stop it from having effects.
Pseudocholinesterase is used in Tx of Alzheimer’s disease.
What are properties of muscarinic cholinergic receptors?
- G-protein coupled receptors (Gq/11 or Gi/o)
- 5 distinct subtypes exist in humans: M1 - M5
- M1/3/5 are stimulatory and coupled to Gq and PLC - increase in Ca/IP3/DAG
- M2/4 are inhibitory and are coupled to adenylyl cyclase and K+ channels, enhancing K+ conductivity (hyperpolarity)
What are properties of nicotinic cholinergic receptors?
- nAChR exhibit direct ligand-gated conductance
- enhance K+ and Na+ conductance equally
- there are 2 subtypes of nAChR:
1. Nm - skeletal muscles and neuromuscular junction
2. Nn - autonomic ganglia, adrenal medulla, and CNS
For the M1 receptor, describe:
1) typical locations
2) responses
3) mechanisms
4) prototype agonist
5) prototype antagonist
1) autonomic ganglia and CNS
2) late excitatory postsynaptic potential (EPSP), complex: at least arousal, attention, analgesia
3) Gq/11 –> PLC –> increase IP3 + DAG –> increase Ca2+ and PKC
4) agonist: oxotremorine
5) antagonist: pirenzepine
For the M2 receptor, describe:
1) typical locations
2) responses
3) mechanisms
4) prototype agonist
5) prototype antagonist
1) heart SA node, AV node, atrium and ventricle
2) slowed spontaneous depolarization; hyperpolarization, decreased conduction velocity, decreased refractory period, decreased contractility force, slight decrease in contractility
3) Beta-gamma portions of G protein inhibit AC and increase K+ channel opening
4) agonist: -
5) antagonist: AF-DX 117
For the M3 receptor, describe:
1) typical locations
2) responses
3) mechanisms
4) prototype agonist
5) prototype antagonist
1) location: smooth muscle
2) responses: contraction
3) mechanism: same as M1 (Gq/11 –> PLC)
4) agonist: -
5) antagonist: hexahydrosiladifenidol
For the M4 receptor, describe:
1) typical locations
2) responses
3) mechanisms
4) prototype agonist
5) prototype antagonist
1) location: CNS
2) responses: -
3) mechanism: same as M2 (Gi)
4) agonist: -
5) antagonist: Himbacine
For the M5 receptor, describe:
1) typical locations
2) responses
3) mechanisms
4) prototype agonist
5) prototype antagonist
1) location: CNS
2) responses: -
3) mechanism: same as M1 (Gq/11)
4) agonist: -
5) antagonist: -
For the Nm receptor, describe:
1) typical locations
2) responses
3) mechanisms
4) prototype agonist
5) prototype antagonist
1) location: skeletal muscle at NMJ
2) response: end-plate depolarization; skeletal muscle contraction
3) mechanism: opening of Na+/K+ channels
4) agonist: phenyltrimethylammonium
5) antagonist: tubocurare
For Nn receptor, describe:
1) typical locations
2) responses
3) mechanisms
4) prototype agonist
5) prototype antagonist
1) locations: autonomic ganglia, adrenal medulla, CNS
2) response: depolarization and firing of postganglionic neuron, secretion of catecholamines, complex: at least arousl, attention, analgesia
3) opening of Na+/K+ channels
4) agonist: dimethylphenylpiperazinium
5) antagonist: trimethylaphan
Where do muscarinic receptors dominate in the CNS?
brainstem and spinal cord
Where do nicotinic receptors dominate in the CNS?
substantia nigra, locus coeruleus, and septum
What are 6 locations where both muscarinic and nicotinic receptors are found in the CNS?
- corpus striatum
- cerebral cortex
- hippocampus
- thalamus
- hypothalamus
- cerebellum
What are 5 major cholinergic projection centers?
- septum
- nucleus basalis
- diagonal band
- laterodorsal tegmentum
- pendiculopontine tegmentum
Because of their location in this region of the brain, what neurologic disease can cholinergic agents be used as Tx for relief?
Cholinergic agents can be used for relief of Parkinson’s Sx because the substantia nigra is packed with nicotinic receptors
What are 3 locations of nicotinic receptors in the peripheral nervous system?
- autonomic ganglia
- adrenal medulla
- neuromuscular junction
How do the subunits of Nn receptors differ from Nm receptors?
Nn receptors are composed only of alpha and beta subunits, whereas Nm is comprised of alpha2-beta-epsilon-gamma (major) or alpha2-beta-gamma-delta