Neurotransmitter Receptors in the Postsynaptic Neuron

F Anne Stephenson, University of London, London, UK Lynda M Hawkins, National Institute of Health, Bethesda, USA
The neurotransmitter receptor proteins are integral components in the communication between adjacent cells of the nervous system, spanning the width of the postsynaptic membrane and protruding into the synaptic cleft and cell cytoplasm. They mediate the effects of neurotransmitters between adjacent neurons.

Introductory article
Article Contents
. Introduction . Interaction of Neurotransmitters with Specific Postsynaptic Receptor Proteins . Specialized Set of Postsynaptic Receptor Proteins for each Neurotransmitter . More than 50 Distinct Neurotransmitter Receptor Types . Rapid Synaptic Transmission via Ion Channel-coupled Neurotransmitter Receptors . Biochemical Signalling at Synapses via Metabotropic Neurotransmitter Receptors . Activation of Both Directly Coupled and Metabotropic Receptors by Acetylcholine and Amino Acid Neurotransmitters . Activation of Mainly Metabotropic Receptors by Small Peptides and Biogenic Amine Neurotransmitters . Signalling and Encoding Abilities of a Synapse Defined by its Complement of Specific Receptor Proteins . Summary

Introduction
Communication between adjacent neurons in the central nervous system occurs at specialized regions of the nerve cells termed the synapse. It is mediated by the movement of chemical mediators or neurotransmitters across a small gap that exists between the nerve cells. When a neuron is activated, it releases molecules of neurotransmitter from synaptic vesicles localized in the presynaptic terminal by a Ca2 1 -dependent exocytotic mechanism. The released neurotransmitter diuses across the gap between the neurons, the synaptic cleft, to the postsynaptic membrane. There are many dierent types of neurotransmitters. They are mostly small hydrophilic molecules such as biogenic amines, e.g. noradrenaline, dopamine and serotonin (also known as 5-hydroxytrytophan or 5-HT); amino acids, e.g. g-aminobutyric acid (GABA), glycine and l-glutamate and low-molecular weight peptides, e.g. enkephalin, substance P, neurotensin, etc. Neurotransmitters are unable to cross or permeate passively through the hydrophobic neuronal postsynaptic membrane. Instead, they mediate their eects by the interaction with a receptor embedded in the postsynaptic membrane, the neurotransmitter receptor. This article describes the properties of this important class of receptor.

binding site on the neurotransmitter receptor and subsequent removal from the synaptic cleft by either an Na 1 dependent transporter protein or by enzymatic inactivation, as occurs for the hydrolysis of acetylcholine by acetylcholinesterase. This is summarized in Figure 1.

Presynaptic terminal

Neurotransmitter transporters

Neurotransmitter

Interaction of Neurotransmitters with Specific Postsynaptic Receptor Proteins

Neurotransmitter receptors are integral membrane proteins, i.e. they span the width of the postsynaptic membrane and protrude both into the synaptic cleft and into the cell cytoplasm. The interaction of the neurotransmitter with the neurotransmitter receptor is a noncovalent, reversible interaction resulting in a conformational change in the receptor protein leading to either an alteration in the permeability of certain ions through the membrane or the activation of intracellular enzymes (see below). Neurotransmitter action is terminated by diusion away from its
Ligand-gated ion channels

Effector

G protein-coupled receptors Postsynaptic neuron

Neurotransmitter receptorassociated proteins

Figure 1 A typical synapse in the central nervous system.

ENCYCLOPEDIA OF LIFE SCIENCES 2001, John Wiley & Sons, Ltd. www.els.net

Neurotransmitter Receptors in the Postsynaptic Neuron

Specialized Set of Postsynaptic Receptor Proteins for each Neurotransmitter

Initially, it was thought that each neurotransmitter interacted with a single type of receptor; thus, for example, the excitatory amino acid neurotransmitter, l -glutamate, mediates neurotransmission by binding with high anity to the l-glutamate receptor. However, the advent of molecular cloning revolutionized this original concept and it is now clear that there exists a family of neurotransmitter receptors for each neurotransmitter. Thus, l-glutamate is now known to be the endogenous activator of the family of glutamate receptors. The dierent members of a family of neurotransmitter receptors are encoded by separate genes; thus, they are distinct proteins. However, these dierent proteins share a high degree of amino acid sequence similarity. Receptors that are ion channels are multisubunit proteins; that is, they are formed by the coassembly or coassociation of the receptor gene products. Neurotransmitter receptors whose transduction mechanism is intracellular enzyme activation, i.e. metabotropic or G protein-coupled receptors, are mostly single-subunit proteins. Ion channel receptors have the potential for the greatest number of dierent receptor subtypes, which is called receptor diversity. This is because, for a xed number of receptor genes, there are many dierent ways that the gene products can coassemble to form a functional receptor. For metabotropic receptors, the number of dierent receptors within one family is dependent only on the number of receptor genes.
R + L* Free kD = k 1 k1

k1 k 1 RL* Bound

Figure 2 The principles of radioligand-binding assays. The receptor (R) preparation and radioligand (L) are incubated to equilibrium. The bound radioactivity is then separated from the free, most frequently by rapid filtration. The bound radioactivity thus yields a measure of the receptor present. Using saturation-binding curves and Scatchard transformation, the affinity of the radioligand for the receptor can be determined.

to that of the receptor-containing tissue: at equilibrium, the bound radioactivity, i.e. a measure of the amount of receptor, can be readily separated from the free ligand by molecular size fractionation. This may be either ltration through glass bre lters or centrifugation. Data generated by these assays can be transformed using the Scatchard plot, which permits the determination of the number of receptors, referred to as Bmax, and the dissociation constant, Kd, i.e. the strength of the binding between the radioactive ligand and receptor. Figure 2 shows the principles of the radioligand-binding assay. Alternatively, the dierent receptor families can be characterized by virtue of their respective transduction mechanisms using either electrophysiological methodology (ion channel receptors) or by enzyme activation (G protein-coupled receptors).

More than 50 Distinct Neurotransmitter Rapid Synaptic Transmission via Ion Channel-coupled Neurotransmitter Receptor Types Receptors
There are over 50 dierent neurotransmitters and each neurotransmitter binds to its own family of neurotransmitter receptors. Because of this selectivity, neurotransmitter receptors are primary targets for pharmacological intervention. Drugs that mimic the action of the neurotransmitter are receptor agonists, whereas drugs that bind to the same site as the neurotransmitter but do not result in receptor activation are receptor antagonists. Some drugs can also discriminate between subtypes of the same receptor family. Neurotransmitter receptors are detected and characterized by radioligand binding assays. These assays involve the incubation to equilibrium of a high-anity, radioactive ligand with a tissue preparation, usually membranes, containing the receptor. The radioactive ligand could be either the natural neurotransmitter or a synthetic agonist or antagonist specic for the receptor under study. Generally, this ligand is of low molecular weight compared
2

As mentioned above, neurotransmitter receptors can be divided into two broad classes based on their transduction mechanisms following receptor activation. Rapid synaptic transmission is mediated by neurotransmitter receptors that are ligand-gated ion channels. The interaction of the neurotransmitter with the respective neurotransmitter receptor for a ligand-gated ion channel results, within milliseconds, in the opening of either a cation- or anionselective integral ion channel. Depending on the selectivity of the ion channel and the membrane resting potential, this results in depolarization or hyperpolarization of the recipient neuron. Prolonged exposure to the neurotransmitter leads to a waning of the conductance changes, i.e. desensitization. Examples of neurotransmitters that gate ion channels are, for excitatory responses, acetylcholine, glutamate, serotonin (5-HT) and adenosine triphosphate (ATP), and, for inhibitory responses, g-aminobutyric acid

Neurotransmitter Receptors in the Postsynaptic Neuron

(GABA) and glycine. Generally, depolarization results from the gating of cation channels, whereas hyperpolarization is the result of the opening of anion channels. But note that GABA can mediate excitatory responses in neonatal hippocampal neurons but this is probably due to the unusual resting potentials in these cells. Also, all the ionotropic glutamate receptors are permeable to Ca 2 1 except those receptors that contain the RNA edited form of the GluR2 subunit. Nicotinic acetylcholine, 5-HT3, GABAA, GABAC and glycine receptors all belong to the same superfamily of ligand-gated ion channels, of which the best characterized is the peripheral nicotinic acetylcholine receptor (nAChR), which is expressed at the neuromuscular junction. All members of this family share structural features and signicant amino acid sequence homologies, even though

acetylcholine and 5-HT gate cation channels and GABA and glycine receptors gate anion channels. They are all pentamers, i.e. they are formed from the coassembly of ve polypeptide chains with molecular weights in the region  4070 kDa. They have four transmembrane domains in each polypeptide, TM1TM4; the TM2 transmembrane domain forms the inner lining of the channel. A further common feature is a large N-terminal extracellular domain, which contains a conserved cys-cys b loop structure whose function is unknown. The C-terminal region of the subunits is also located extracellularly. nAChRs are anchored and clustered in the postsynaptic membrane by the protein, rapsyn. Dierent proteins have similar roles for the other ligand-gated ion channels. Figure 3a summarizes the pertinent features of the nAChR ligand-gated ion channel superfamily.

NT C C C 192 C 193 CT V In 1 Out 2 Acetylcholine binding sites

(a)

(c)

Na+ NT

ACh+

K+

Out 110 In CT

80 (b) (d) Rapsyn

Figure 3 Pertinent features of ligand-gated ion channel neurotransmitter receptors. (a) The key features, including the transmembrane topology of nicotinic acetylcholine, g-aminobutyric acid (GABAA), glycine and serotonin (5-HT3) receptors. (b) The key features of the ionotropic glutamate receptors. NT, N-terminal; CT, C-terminal; I, II, III, IV are the membrane domains; the red arrows point to the sites of N-glycosylation; C-C, the cys-cys loop. (c) The arrangement of the subunits and the channel pore of the nicotinic acetylcholine receptor as viewed perpendicular to the plane of the membrane. (d) The nicotinic acetylcholine (ACh) receptor in the membrane together with the acetylcholine-binding site (drawn to scale). (d) Reproduced in part from resolution. Journal of Molecular Biology 288: 765 786, by permission of the publisher, Miyasawa A et al. (1999) Nicotinic acetylcholine receptor at 4.6 A Academic Press.

Neurotransmitter Receptors in the Postsynaptic Neuron

The quaternary structure of ionotropic glutamate receptors is uncertain at the present time. However, it is known that they are heteromeric, with functional receptors being composed of four or ve polypeptide chains. These subunits are all larger than those found for members of the original ligand-gated ion channel superfamily, with molecular weights in the range 120180 kDa. Each has four membrane domains but, in contrast to the receptors described above, only three of these span the membrane. The second membrane domain is monotropic, i.e. it does not span the lipid bilayer but it does form the inner lining of the channel lumen. The C-terminal portion of the subunit is intracellular. The ionotropic glutamate receptor subunits do not contain the cys-cys b-loop structure, nor do they share any amino acid sequence similarity with nAChRs, GABAA/C or glycine receptors. ATP-gated neurotransmitter receptors are termed P2X receptors. They are also multisubunit proteins but they are dierent to both the nAChR and glutamate receptor families. There are seven dierent P2X subunits, which can assemble as either homomultimers or heteromultimers to form functional channels. The number of subunits in each channel molecule is not known. P2X subunits each have two transmembrane domains that contribute to the formation of the ion channel; both the N- and the Ctermini of each subunit are located intracellularly. The properties of ligand-gated ion channels are summarized in Table 1.

Biochemical Signalling at Synapses via Metabotropic Neurotransmitter Receptors

Neurotransmitter receptors that mediate slower responses at the synapse, of the order of seconds, are metabotropic or alternatively, G protein-coupled receptors. For this

receptor family, the signalling mechanism consists of a receptor protein, a guanine nucleotide protein (G protein) and an eector molecule, which is an enzyme that catalyses the production of intracellular messengers. Metabotropic receptors are the largest family of neurotransmitter receptor. The prototype is the b-adrenergic receptor, which itself has structural (but not amino acid sequence!) homology to the protein bacteriorhodopsin, and homology with rhodopsin, the light-harvesting protein expressed in the eye. Metabotropic neurotransmitter receptors are, mostly, single subunit receptors. Each receptor has seven transmembrane-spanning domains, an extracellular Nterminus and an intracellular C-terminus, which for some receptors is anchored to the cytoplasmic face of the membrane by palmitoylation (Figure 4). Receptor heterogeneity in this receptor family is created by the existence of separate genes for a particular neurotransmitter receptor. Additional diversity is created by the coupling of dierent receptor subtypes to dierent G proteins, hence dierent enzyme eectors. For example, b1-, b2-, b3- and b4adrenergic receptors can couple to Gs proteins and activate adenylate cyclase, thus increasing the intracellular concentration of cyclic adenosine monophosphate (cAMP); a1A, a1B and a1D-adrenergic receptors mediate their responses via coupling to Gp/Gq and subsequent activation of phospholipase C, resulting in an increase in the second messengers, diacylglycerol and inositol trisphosphate; a2A, a2B- and a2D-adrenergic receptors are negatively coupled to adenylate cyclase via Gi/GO. Although all members of this family of metabotropic neurotransmitter receptors have the characteristic seven transmembrane domains, the family can be subdivided on the basis of amino acid sequence homologies. Group 1 receptors are 300400 amino acids in length, they have a small N-terminal extracellular region, generally bind small ligands, such as adrenaline, noradrenaline, dopamine, serotonin, opiates and tachykinins, and have the highest degree of amino acid sequence similarity within the

Table 1 Ligand-gated ion channel neurotransmitter receptors Neurotransmitter Acetylcholine Adenosine triphosphate Glutamate Receptor type Nicotinic (peripheral) Nicotinic (neuronal) P2X17 NMDA AMPA Kainate GABAA GABAC GlyR 5-HT3 Channel selectivity Na 1 /K 1 /Ca2 1 Na+/K+/Ca2+ Na 1 /K 1 /Ca2 1 Na 1 /K 1 /Ca2 1 Na+/K+/Ca2+ Na+/K+/Ca2+ Cl 2 Cl 2 Cl 2 Na 1 /K 1 /Ca2 1

g-Aminobutyric acid Glycine 5-Hydroxytrypamine

AMPA, a-amino-3-hydroxy-5-methylisoxazolepropionate; NMDA, N-methyl-d -aspartate.

Neurotransmitter Receptors in the Postsynaptic Neuron

NT

Out In (a) CT

NT

V V V Out V Neurotransmitter binding site

transmembrane regions. Group 2 metabotropic receptors are signicantly larger, with 9001200 amino acids; they have large N-terminal and C-terminal domains and, signicantly, they do not share amino acid sequence similarity with group 1 receptors. Metabotropic glutamate receptors and GABAB receptors belong to this class. Interestingly, GABAB receptors are known to be heterodimers. Groups 3 and 4 also exist but, generally, they are not receptors for neurotransmitters but hormones. Nonpeptide neurotransmitters bind to their respective receptors in a cavity within the membrane formed by the seven transmembrane regions. Residues in the extracellular regions have been implicated in the binding of peptide ligands. The dierent types of adrenergic receptor, together with their transduction mechanisms, are summarized in Table 2. Table 3 lists additional examples of metabotropic neurotransmitter receptors.

In G protein-binding site CT

Activation of Both Directly Coupled and Metabotropic Receptors by Acetylcholine and Amino Acid Neurotransmitters
The neurotransmitters acetylcholine, GABA, l-glutamate and 5-HT are unusual, in that they activate both ionotropic and metabotropic receptors. Thus acetylcholine activates either nicotinic acetylcholine receptors, which are ion channels, or muscarinic acetylcholine receptors, which belong to the G protein-receptor family. Similarly GABAA/C and ionotropic l-glutamate receptors are ion channels, whereas GABAB and metabotropic l-glutamate receptors are G protein-coupled receptors. In each case, there is no structural or amino acid sequence similarity between the ionotropic and metabotropic receptors activated by the same neurotransmitter. Further, their respective pharmacological proles are dierent.

Figure 4 Pertinent features of G protein-coupled neurotransmitter receptors. (a) The key features, including the transmembrane topology of metabotropic G protein-coupled receptors. NT, N-terminal; CT, Cterminal; the solid rectangles are the seven transmembrane domains; the red arrows point to the sites of N-glycosylation and the zig-zag line represents the palmitoylation at the C-terminus. (b) The G protein-coupled receptor as viewed through the plane of the membrane.

Table 2 Adrenergic receptor subtypes: an example of a G protein-coupled neurotransmitter receptor which has multiple subtypes that couple to a variety of G proteins, resulting in activation of dierent intracellular eector molecules Adrenergic receptor subtype a1A, a1B, a1D a2A, a2B, a2C b1 b4 G protein Gq/11 Gi/o Gs Transduction mechanism "IP3/DAG #cAMP "cAMP

IP3, inositol 1,4,5- trisphosphate; DAG, diacylglycerol; cAMP, adenosine 3,5-cyclic monophosphate.

Neurotransmitter Receptors in the Postsynaptic Neuron

Table 3 G protein-coupled neurotransmitter receptors demonstrating receptor multiplicity and diversity with respect to transduction mechanism Neurotransmitter Acetylcholine Adenosine triphosphate Dopamine Receptor subtype Muscarinic M1, 3, 5 Muscarinic M2, 4 P2Y D1, D5 D2 D3, D4 GABAB (GBR1, 2) mGluR1, 5 mGluR2, 3, 4, 6, 7, 8 5-HT1A, B, D, E, F 5-HT2A, B, C 5-HT4, 5, 6, 7 SST1-5 G protein Gq/11 Gi/o Gq Gs Gi/o ? Gi/o Gq/11 Gi/os Gi/Go Gq/11 Gs Gi/o Transduction mechanism "IP3/DAG #cAMP "IP3/DAG "cAMP #cAMP ? #cAMP "IP3/DAG #cAMP #cAMP "IP3/DAG "cAMP #cAMP

g-Aminobutyric acid Glutamate 5-Hydroxytryptamine

Somatostatin

IP3, inositol 1,4,5-trisphosphate; DAG, diacylglycerol; cAMP, adenosine 3,5-cyclic monophosphate.

Activation of Mainly Metabotropic Receptors by Small Peptides and Biogenic Amine Neurotransmitters
In Tables 1, 2 and 3, it can be seen that, generally, metabotropic receptors are activated by low-molecular weight peptides and biogenic amines, whereas the fastacting ionotropic receptors are activated predominantly by amino acid neurotransmitters, the noted exception being acetylcholine. Peptides are often coreleased with fastacting neurotransmitters, thus implying a modulatory role following interaction with the slower-acting G proteincoupled receptors.

Presynaptic Group II m Glu R 2/3

Presynaptic active zone

AMPA

Group m Glu R 4/7/8 2 NMDA AMPA Postsynaptic specialization m Glu R 5

Signalling and Encoding Abilities of a Synapse Defined by its Complement of Specific Receptor Proteins
The mammalian neuromuscular junction is a specialized synapse as it receives only excitatory input from the neurotransmitter, acetylcholine, which interacts with a homogeneous population of receptors, ligand-gated nicotinic acetylcholine receptors. Receptor activation always results in depolarization of the muscle cell. In the central nervous system, synapses are more complex. Each synapse generally receives either an inhibitory or an excitatory input from a single type of neurotransmitter; hence, synapses are referred to as GABAergic, where GABA is the neurotransmitter, glutamatergic for l-glutamate, etc. Some synaptic terminals may release more than one neurotransmitter, thus multiple corresponding neuro6

Cerebellum

Hippocampus

Postsynaptic
Figure 5 Organization of the different types of glutamate receptor at glutamatergic synapses. The diagram summarizes information that has accrued from immunochemical studies of hippocampal and cerebellar glutamatergic synapses, as indicated, using the electron microscope. Up to four different types of glutamate receptors are found in the postsynaptic membrane, including both ionotropic and metabotropic receptors. Thus it can be seen that the signalling and encoding abilities of a synapse must be defined by the complement of the receptor proteins colocalized at these synapses. AMPA, a-amino-3-hydroxy-5-methylisoxazoleproprionate; NMDA, N-methyl-D-aspartate. Reproduced with permission from Takumi Y et al. (1998) Synaptic arrangement of glutamate receptors. Progress in Brain Research 116: 105 121, by permission of the publisher, Elsevier Press.

Ext ras me ynap mb tic ran e

m Glu R1

Perisynaptic annulus

ptic yna e ras n Ext mbra me

Neurotransmitter Receptors in the Postsynaptic Neuron

transmitter receptors may coexist within the postsynaptic neuron; however, even at synapses releasing a single neurotransmitter, the receptors can be heterogeneous with respect to both the type and subtype of receptor. Thus, the overall response at the synapse will be a summation or integration of all the receptor responses. For example, at a glutamatergic synapse, in the postsynaptic membrane metabotropic, non-NMDA (N-methyl-d -aspartate) and NMDA types of glutamate receptors are found. For each of these, subtypes may coexist. Figure 5 summarizes the organization of glutamate receptors at a typical glutamatergic synapse.

ionotropic, or they may be metabotropic receptors. This major class of neurotransmitter receptor is slower in action because it involves association with the transducer, the appropriate G protein, followed by intracellular enzyme activation. Many subtypes of neurotransmitter receptor may be found at a single synapse in the central nervous system; thus, the overall response of a synapse will be dened by its complement of specic receptor proteins. Other articles describe in more detail the structures and pharmacological properties of the neurotransmitter receptors, together with more in depth descriptions of their respective downstream signalling pathways.

Summary
There are many dierent types of neurotransmitter molecules which mediate the communication between adjacent neurons via interaction with integral membrane proteins, the neurotransmitter receptors. These neurotransmitter receptors can be either fast acting, i.e.

Further Reading
Levitan IB and Kaczmarek LK (1997) The Neuron: Cell and Molecular Biology, 2nd edn. New York: Oxford University Press. Stephenson FA and Turner AJ (eds) (1998) Amino Acid Neurotransmission. London: Portland Press. Zigmond MJ, Bloom FE, Landis SC, Roberts JL and Squire LR (eds) (1998) Fundamental Neuroscience. London: Academic Press.