Phar 590/735 - Winter 2006 Dr. Jane Ishmael Office: Phar 411 Phone: 737-5783 E mail: Jane.Ishmael@oregonstate.edu.

SIGNAL TRANSDUCTION THE LIGAND-GATED ION CHANNELS

1.
BACKGROUND

Three Types of Transmembrane Channel: 1. Ligand-gated Ligand binds to ion channel alters ion conductance 2. Voltage-gated Change in transmembrane voltage gradient alters ion conductance 3. Second messenger regulated Binding of a ligand to a G-protein coupled receptor generates a second messenger that regulates ion conductance of a channel

Neurons communicate with each other at synapses

Neurons signal by transmitting electrical signals (action potentials) along their axons (0.1 mm to 3 m) Action potentials are initiated at the initial segment of the axon and are conducted down the axon at rates of 1 100m / sec Many axons are insulated by a myelin sheath to increase the speed of transmission (many are not insulated)

Electrical signals flow in one direction (action potential is propagated unidirectionally) and make specific connections with postsynaptic target cells (i.e networks are not random)
At the end of the axon, voltage changes trigger the release of neurotransmitters Drug selectivity is based on the fact that different neuronal pathways utilize different neurotransmitters It has been estimated that one neuron communicates with 1000 others!

ION CHANNELS MEDIATE FAST SIGNALLING EVENTS Speeds of around one millisecond G PROTEIN-COUPLED RECEPTORS (GPCR) use mechanisms that are at least one hundred times slower TRANSMEMBRANE RECEPTORS WITH CYTOSOLIC DOMAINS 100s millisecs to minutes STEROID/HORMONE FAMILY of NUCLEAR RECEPTORS Respond in minutes to hours

Note that in many cases MORE THAN ONE TYPE of RECEPTOR has evolved to recognize a specific ligand/neurotransmitter e.g GABA, glutamate, serotonin, acetylcholine use GPCRs and ligand-gated ion channels

BASIC CHARACTERISTICS of ligandactivated channels

LIGAND GATED = NEUROTRANSMITTER ACTIVATED

Ligand-gated ion channels are transmembrane proteins. Comprised of multiple subunits that assemble in the membrane to form a central pore. WHEN NEUROTRANSMITTER binds the PROTEIN undergoes a CONFORMATIONAL CHANGE The CONFORMATIONAL CHANGE OPENS a PORE ACROSS the PLASMA MEMBRANE The STRUCTURE of the protein determines which IONS flow into or out of the cell through the pore resulting in: o EXCITATION or o INHIBITION

A conformational change INACTIVATES the CHANNEL Further conformational change resets the channel to a CLOSED state i.e. ready to be activated

THERE ARE SEVERAL SUPERFAMILIES OF LIGAND-GATED IN CHANNELS

CYS-LOOP SUPERFAMILY

5 subunits make 1 receptor Each subunit has 4 transmembrane domains

Two distinct groups: 4.1 CATIONIC RECEPTORS Nicotinic Acetylcholine Receptors 5-HT3 Receptors 4.2 ANIONIC RECEPTORS GABAA Glycine

4.3 Nicotinic Acetylcholine Receptor

REVIEW pages 71 to 75 (Chapter 6) of the required, Principles of Pharmacology Text.
Mediates fast neurotransmission at the neuromuscular junction, peripheral autonomic ganglia and the CNS

4.4 GABA and Glycine-activated channels

GABA (g-aminobutyric acid) and Glycine are inhibitory neurotransmitters

GABA is the major inhibitory neurotransmitter in brain Glycine is the major inhibitory neurotransmitter in spinal cord and brainstem (trace amounts of GABA are found in the periphery). Antagonists of inhibitory neurotransmitters cause convulsions

4.4.1

GABAA receptor

Heterooligomeric protein, composed of 5 subunits that span the cell membrane to form a chloride channel Multiple subtypes of each subunit GABA binds to the extracellular surface channel opens allowing Cl_ ions flow down their concentration gradient hyperpolarization of postsynaptic neuronal membrane

10

4.4.2

Glycine Receptor

High concentrations in the spinal cord and brain stem Inhibitory neurotransmitter Chloride channel Oligomeric transmembrane protein comprised of 5 subunits Strychnine is a competitive glycine antagonist (powerful convulsant) Strychnine-sensitive inhibitory glycine receptor

5. Glutamate Receptors
Major excitatory neurotransmitter in the brain. 25-30% of neurons utilize glutamate as a neurotransmitter. 4 subunits make 1 receptor membrane topology is distinct from subunits of the cys loop family

Ionotropic Glutamate receptors

Metabotropic glutamate receptors are GPCR Originally classified pharmacologically based upon activation by synthetic glutamate analogues: (1) NMDA (N-methyl-D-aspartate) (2) non-NMDA: All 3 types of ionotropic glutamate receptor are composed of multiple subunits that assemble to form a central cation channel. Each subunit is encoded by its own gene.

11

Membrane Topology of Glutamate Receptor Subunits

H2N

OUT

IN

COOH

12