ION CHANNEL PROTEINS.

form hydrophilic pores transport inorganic ions generally called ion channels. participate only in passive transport (facilitated diffusion) molecules are transported "downhill" are ion selective and gated (i.e.. open briefly and then close). Stimuli that open gates include 1. Mechanical stress mechanical-gated ion channels 2. Changes in voltage across the cell membrane (voltage-gated ion channels) 3. Ligand binding (ligand-gated ion channels). One important types = neurotransmitter-gated ion channels mediate ion movement. neuotransmitter-gated ion channels may have an 1. Excitatory 2. Inhibitory Some important ion channel proteins include the following:

Voltage-gated ion channels

L-type Ca2+ channel protein L = long-lasting
(dihydropyridine receptor) is an important determinant of vascular tone and cardiac contractility. In skeletal muscle cells, found on the T-tubules action potential T- tubuleschanges its conformation induces the action of the fast Ca2+ channel protein (ryanodine receptor). Skeletal muscle depends mainly on intracellular pools of Ca2+ from SR for contraction and does not require much transmembrane Ca2+ influx through the L-type Ca2+ channel protein. Therefore, skeletal muscle is not significantly affected by Ca2+ channel blockers.

In vascular smooth muscle cells:

L-type Ca2+ channel protein is found on the cell membrane and changes its conformation in response to the depolarization of an action potential to allow transmembrane Ca2+ influx into the cytoplasm and also induces more Ca2+ release from the sarcoplasmic reticulum via the fast Ca2+ release channel protein. Ca2+ binds to calmodulin and the Ca2+-calmodulin complex that activates myosin kinase, which in turn phosphorylates the myosin crossbridge-ADP-PO42- complex. ADP-PO42- complex is released, leaving the phosphorylated myosin crossbridge bound to actin, which initiates vascular smooth muscle contraction.

In Heart, L-type Ca2+

found in the SA nodal cells, AV nodal cells, and ventricular myocytes. In the SA and AV nodes, phase 0 of the slow action potential is caused by Ca2+ influx into nodal cells through the L-type Ca2+ channel protein. In ventricular myocytes, phase 2 of the fast action potential is caused by Ca2+ influx into ventricular myocytes through the L-type Ca2+ channel protein. This transmembrane Ca2+ influx is called "trigger Ca2+" and is involved in myocyte contraction.

Ca2+ channel antagonists (blockers).

Blocking Ca2+ influx through L-type Ca2+ channel protein. Nifedipine and amlodipine o dihydropyridine class of Ca2+ channel blockers o predominately block Ca2+ influx in vascular smooth muscle cells arterial vasodilation Diltiazem = benzothiazepine class of Ca2+ channel blockers. verapamil = phenylalkylamine class of Ca2+ channel blockers Diltiazem and verapamil have two major actions: Act on SA and AV nodal cells ; which depend on Ca2+ currents for phase 0. Slow rise of phase 0 slowed conduction velocity through the AV node. Prolong repolarization of AV node effective refractory period of AV node. Both actions = very effective in re-entrant arrhythmias.

Fast Ca2+ release channel protein = Ryanodine receptor

In skeletal muscle cells:
found on SR and changes its conformation in response to its interaction with the L-type Ca2+ channel protein to allow transmembrane Ca2+ influx into the cytoplasm. The Ca2+ binds to troponin C, which allows the myosin crossbridge-ADP-PO42- complex to bind actin. ADP-PO42- complex is released leaving the myosin crossbridge bound to actin, which initiates skeletal muscle contraction.

In smooth muscle cells;

fast Ca2+ release channel found on the SR and changes its conformation in response to its interaction with the L-type Ca2+ channel protein to allow transmembrane Ca2+ influx into the cytoplasm. The Ca2+ binds to calmodulin and the Ca2+-calmodulin complex that activates myosin kinase, which in turn phosphorylates the myosin crossbridge- ADP-PO42- complex. ADP-PO42- is released, leaving the phosphorylated myosin crossbridge bound to actin which initiates vascular smooth muscle contraction. the fast Ca2+ release channel protein is found on the SR of cardiac myocytes. "Trigger Ca2+" stimulates the release of a large pool of Ca2+ stored in the SR through the fast Ca2+ release channel protein into the cytoplasm. The Ca2+ binds to troponin C which allows the myosin crossbridge- ADP-PO42- complex to bind actin. ADP-PO42- is released leaving the myosin crossbridge bound to actin, which initiates cardiac myocyte contraction.

In the heart

T-type Ca2+ channel protein (T = transient)

In the heart
found on SA and AV nodal cells and plays a role in phase 4 of the action potential. Phase 4 is caused by : 1. Ca2+ influx into nodal cells through the T-type Ca2+ channel protein 2. and Na+ influx into nodal cells through slow (funny) Na+ channels.

In the brain,
found on relay neurons connecting the thalamus to the cortex. In awake state, these neurons faithfully transmit sensory signals to the cortex and the T-type Ca2+ channel protein is depolarized and inactive. In sleep state, the transient, bursting activity of T-type Ca2+ channel alters incoming signals so that output to the cortex has an oscillator firing rate, which has a characteristic "spike and wave" readout on an electroencephalograph (EEG). Absence seizures (petit mal seizure) are associated with paroxysmal hyperpolarization and activation of the T-type Ca2+ channel protein in the awake state. Patients with absence seizures have EEG readings somewhat similar to patterns generated in slowwave (stage 3) sleep. Ethosuximide is a specific T-type Ca2+ channel protein antagonist and is the first line therapy for uncomplicated absence seizures.

K+ channel protein.
These are the most abundant class of ion channel proteins whereby at least 16 types and many more subtypes have been described but their physiologic functions are far from being fully understood. There are voltage-gated K+ channel proteins that include the: 1. delayed rectifier (Kv) 2. inward rectifier (KIR), 3. transient outward rectifier (KA). There are also Ca2+-activated K+ channel proteins which include the: 1. large conductance channel (BKCa), 2. intermediate conductance channel (IK), 3. small conductance channel (SKCa). metabolically gated K+ channel proteins that include: 1. the ATP-sensitive K+ channel (KATP), 2. arachidonic acid modulated K+ channel (KAA), 3. acetylcholine-activate K+ channel (KACh). In the heart, the K+ channel protein is found on SA and AV nodal cells and cardiac myocytes.

In SA and AV nodal cells.

Adenosine stimulates P1 purinergic receptors opening of a G protein-linked K+ channel (possibly KIR or KATP) so that K+ moves from intracellular into the extracellular space resulting in a hyperpolarization. This inhibits the SA nodal, atrial, and AV nodal conduction. The AV node is more sensitive to adenosine than the SA node. Adenosine has a half-life of 10 seconds and is often used as a first line therapy for converting narrowcomplex paroxysmal supraventricular tachycardia to a normal sinus rhythm.

In cardiac myocytes.
Phase 2 (plateau phase) of the action potential in cardiac myocytes is determined by the depolarizing Ca2+ influx through the L-type Ca2+ channel protein and the hyperpolarizing K+ efflux through the K+ ion channel protein. Larger hyperpolarizing K+ currents shorten the phase 2 plateau duration causing the membrane potential to return to its resting value more rapidly. Shorter hyperpolarizing K+ currents lengthen the phase 2 plateau duration causing the membrane potential to return to its resting value more slowly. When K+ channel proteins are blocked, a smaller hyperpolarizing K+ current is generated and therefore cause a longer phase 2 (plateau phase) and prolonged repolarization. On the beneficial side, a longer phase 2 (plateau phase) increases the effective refractory period and decreases the incidence of re-entry. On the toxic side, a longer phase 2 (plateau phase) increases the likelihood of early after-depolarizations and torsades de pointes. Ibutilide is a class III antiarrhythmic K+ channel protein antagonist (blocker) that also enhances a slow Na+ influx that further prolongs repolarization. Sotalol is a mixed class II (nonselectively antagonizes (-adrenergic receptors) and a class III antiarrhythmic K+ channel protein antagonist (blocker). Bretylium is an antihypertensive agent (performs a "chemical sympathetectomy" by inhibiting the release of norepinephrine) and a class III antiarrhythmic K+ channel protein antagonist (blocker). Amiodarone is mainly a class III antiarrhythmic K+ channel protein antagonist (blocker). Amiodarone is also a class I (blocks Na+ channels and therefore decreases the rate of firing of SA and AV nodal cells), class II (noncompetitive antagonist of - and -adrenergic receptors), and class IV (L-type Ca2+ channel protein antagonist) antiarrhythmic.

Slow (funny) Na+ channel protein.

The slow (funny) Na+ channel protein is found on SA and AV nodal cells in the heart and causes a slow, inward current of Na+. Phase 4 of the slow action potential observed in SA and AV nodal cells is caused by Ca 2+ influx into nodal cells through T-type Ca2+ channel proteins and Na+ influx into nodal cells through slow (funny) Na+ channel proteins.

Fast Na + channel protein.

The fast Na+ channel protein is found on cardiac myocytes and causes a fast, inward current of Na+. Phase 0 of the fast action potential observed in cardiac myocytes is caused by Na+ influx into cardiac myocytes through fast Na+ channel proteins. Na+ channel protein antagonists (blockers) decrease the upstroke velocity of phase 0 and thereby decrease the conduction velocity through cardiac tissue. Class IA Na+ channel protein antagonists prolong repolarization and thereby increase the effective refractory period. In addition, Na+ channel protein antagonists decrease the slope of phase 4 depolarization and increase the threshold potential in SA and AV nodal cells and thereby decrease the automaticity in SA nodal cells. Quinidine, procainamide, disopyramide are class IA antiarrhythmic Na+ channel protein antagonists (blockers) that bind preferentially to open Na+ channel proteins. Lidocaine, tocainide, and mexiletine are class IB antiarrhythmic Na+ channel protein antagonists (blockers) that bind preferentially to open and inactivated Na+ channel proteins. A major distinguishing characteristic of class IB antiarrhythmics is their fast dissociation from Na+ channel proteins. Flecainide, propafenone, moricizine, and encainide are class IC antiarrhythmic Na+ channel protein antagonists (blockers), which are the most potent Na+ channel protein blockers.

Transmitted-gated ion channels

Nicotinic acetylcholine (nACh) receptor.
The nACh receptor is composed of two , one , one , and either one or one subunit (i.e., five subunits). The 2 nACh receptor is found at the neuromuscular junction of mature skeletal muscle and the 2 nACh receptor is found at the neuromuscular junction of embryonic skeletal muscle. The -subunits are responsible for binding acetylcholine (ACh), which is the structural basis for the binding of two ACh molecules per receptor. When two molecules of ACh bind to the nACh receptor during a discrete, brief pulse, the gate is opened and the influx of Na+ and Ca2+ and efflux of K+ occurs. The gate remains open for only about 1 msec when it is nonselectively permeable to all cations (but excludes anions). The influx of Na+ (30,000 Na+/nACh receptor/msec) is primarily responsible for the depolarization of the postsynaptic membrane and mediates the excitatory postsynaptic potential (EPSP). During a continuous exposure of nACh receptor to ACh, the nACh receptor undergoes a change to a desensitized conformation whereby the channel is locked closed. nACh receptors are also found in the CNS, autonomic ganglia, and the adrenal medulla where they are termed N2 or NN. nAChRs found in skeletal muscle are termed N1 or NM. Succinylcholine (Anectine, Quelicin, Sucostrin) is a nACh receptor agonist that competes with ACh for the nACh receptor. Succinylcholine maintains an open Na+ channel eventually causing skeletal muscle relaxation and paralysis. Succinylcholine is used to induce paralysis during surgery by means of a depolarizing blockade. Succinylcholine may cause malignant hyperthermia which is a major cause of anesthesia related deaths. Malignant hyperthermia results from an excessive release of Ca2+ from the sarcoplasmic reticulum and presents as hyperthermia, metabolic acidosis, tachycardia, and accelerated muscle contractions. Treatment includes: rapid cooling, 100% oxygen, control of acidosis, and administration of Dantrolene, which blocks release of Ca2+ from the sarcoplasmic reticulum. Nicotine is considered a nACh receptor agonist because it binds to ACh-binding sites and activates the nACh receptor. Nicotine may be responsible for the psychophysical effects of smoking addiction by its interaction with N2 or NN nACh receptors in the CNS. Tubocurarine, pancuronium, vecuronium, and rocuronium are nACh receptor antagonists (blockers) that block endogenous ACh binding. These blockers are used to induce paralysis during surgery by means of a nondepolarizing blockade. Because nACh receptors (i.e., N2 or NN) are found on autonomic ganglia, these blockers often have side effects associated with ganglionic blockade, which can be reversed by acetylcholinesterase inhibitors (e.g., neostigmine).

Glutamate receptors.
Glutamate is the major excitatory neurotransmitter in the CNS and elicits its action by binding to glutarnate receptors. There are two main classes of glutarnate receptors: 1. transmitter-gated ion channel proteins (discussed here) and 2. G protein-linked receptors (sometimes called metabotropic receptors will be discussed later). The glutamate-gated ion channel proteins include the following: N-methyl-D-aspartate (NMDA) Receptor. The NMD A receptor is found throughout the CNS (e.g., hippocampus, cerebral cortex, and spinal cord). When glutarnate and the cofactor glycine bind to the NMDA receptor, the gate is opened and the influx of Na+ and Ca2+ and efflux of K+ occurs. In addition, a voltage-dependent membrane depolarization is required to open the gate because the NMDA receptor is "plugged" by extracellular Mg2+ at resting membrane potential. Amantadine is a noncompetitive NMDA receptor antagonist (blocker). Memantine is a noncompetitive, use-dependent NMDA receptor antagonist. Ketamine (Ketalar; Special K; K) is an NMDA receptor antagonist that acts as a CNS depressant and a dissociative anesthetic. Ketamine has sedative, analgesic, and hallucinogenic effect and is used as a "date rape" drug. Phencyclidine (PCP; angel dust) and dizocilpine (MK-801) are NMDA receptor antagonists that cause hallucinogenic behavior and require the NMDA receptor to be open to gain access to their binding sites (i.e., open channel blockers). Kainate receptor. The kainate receptor is found throughout the CNS (e.g., hippocampus, cerebellum). When glutarnate binds to the kainate receptor, the gate is opened and the influx of Na+ and the efflux of K+ occurs. -amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor. The AMPA receptor is found throughout the CNS (e.g., hippocampus, cerebral cortex). When glutarnate binds to the AMPA receptor, the gate is opened and the influx of Na+ and Ca2+ and the efflux of K+ occurs. Ca2+ influx through AMPA receptors may play a role in neuronal cell death. Excitotoxicity refers to neuronal cell death caused by toxic levels of glutamatergic neurotransmission, which increase neuronal Ca2+ levels. Some diseases associated with excitotoxicity include CNS ischemia, physical trauma, epilepsy, Parkinson disease, amyotrophic lateral sclerosis (ALS), and Alzheimer disease.

5-Hydroxytryptamine3 (5-HT3) serotonin receptor. When serotonin binds to the 5-HT3 receptor, the gate is opened and the influx and Na+ and efflux of K+ occurs. The 5-HT3 receptor belongs to a lamily of 5-HT receptors whose other members are G protein-linked receptors, which will be discussed later.

Purinergic2x (P2X) receptor. Adenosine and ATP are neurotransmitters that elicit their action by binding to purinergic receptors. The P2X receptor is found on peripheral terminals of nociceptive neurons and is involved in the sensation of acute pain caused by tissue damage and inflammation (i.e., nociception). When adenosine or ATP binds to the P2X receptor, the gate is opened and theinflux and Na+ and Ca2+ and efflux of K+ occurs. The P2X receptor belongs to a family of purinergic receptors whose other members are G proteinlinked receptors, which will be discussed later. Gamma-aminobutyric acidA (GABAA) Receptors. GABA is the major inhibitory neurotransmitter in the CNS and elicits its action by binding to GABA receptors. There are two main classes of GABA receptors: transmitter-gated ion channel proteins (GABAA receptor discussed here) and G protein-linked receptors (GABAB receptor, sometimes called metabotropic receptors,will be discussed later). The GABAA receptor is found in most areas of the CNS and generates a fast inhibitory postsynaptic potentials (IPSPs). The GABAA receptor is composed of two a subunits and three (3 or 7 subunits (i.e., five subunits). The a subunits are responsible for binding GABA which is the structural basis for the binding of two GABA molecules per receptor. When two molecules of GABA bind to the GABAA receptor, the gate is opened and the influx of Cl~ occurs (causes hyperpolarization of the cell). Muscimol is a GABAA receptor agonist. Bicuculline is a GABAA receptor competitive antagonist. Picrotoxin is a GABAA receptor noncompetitive antagonist. The drugs used in clinical practice (i.e., barbiturates and benzodiazepines) to effect the GABAA receptor fall into the category of GABAA receptor modulators. Barbiturates. Thiopental, methohexital, pentobarbital, secobarbital, amobarbital, and phenobarbital are GABAX receptor modulators that enhance the efficacy of GABA by increasing the duration of CI channel opening leading to membrane hyperpolarization and decreased neuronal excitability.

Benzodiazepines. Midazolam, clorazepate, alprazolam, lorazepam, chlor-diazepoxide, clonazepam, diazepam, triazolam, estazolam, temazepam, flurazepam, and quazepam are GABAA receptor modulators that enhance the potency of GABA by increasing the frequency of CI- channel opening leading to membrane hyperpolarization and decreased neuronal excitability. Flunitrazepam (rohypnol; "roofies"; Forget Me Pill) is a fast-acting benzodiazepine that causes amnesia and is used as a "date rape" drug. Glycine receptor. The amino acid glycine is the major inhibitory neurotransmitter in the spinal cord and elicits its action by binding to glycinereceptors. The glycine receptor is composed of -subunits and -subunits. When glycine binds to the glycine receptor, the gate is opened and the influx of CI- occurs (causes hyperpolarization of the cell).