PERTUSSIS TOXIN-SENSITIVE GUANINE NUCLEOTIDE-BINDING PROTEIN(S) COUPLE ADENOSINE A1 AND 5-HYDROXYTRYPTAMINE1A RECEPTORS TO THE SAME EFFECTOR SYSTEMS IN RAT HIPPOCAMPUS - BIOCHEMICAL AND ELECTROPHYSIOLOGICAL STUDIES

  • 1 April 1989
    • journal article
    • research article
    • Vol. 35  (4) , 484-494
Abstract
Distinct membrane receptors that elicit similar cellular responses may share elements of signal transduction. In the present study, rat hippocampal adenosine (AD) and 5-hydroxytryptamine (5-HT) receptors were chosen to test this possibility using biochemical and electrophysiological techniques. Responses elicited by the AD receptor that mediates the inhibition of forskolin-stimulated adenylyl cyclase activity in rat hippocampal membranes and hyperpolarization of resting membrane potential (RMP) in rat hippocampal pyramidal cells were characterized and compared, in the same preparation, with those analogous responses elicited by the 5-HT1A receptor. A series of AD agonists including the selective AD A1 agonist (R)-phenylisopropyladenosine [(R)-PIA]inhibited forskolin-stimulated adenylyl cyclase activity in rat hippocampal membranes in a concentration-dependent manner. Cyclopentyltheophylline (CPT), a selective AD A1 antagonist, was a potent, competitive antagonist of this response with a dissociation constant (Kb) of 6 nM (Schild analysis). The rank order of agonist EC50 values and antagonist Kb values, as well as stereoselectivity, are consistent with the classification of this receptor as the AD A1 receptor. Spiperone, a potent 5-HT1A antagonist, competitively antagonized 5-HT-mediated inhibition of forskolin-stimulated adenylyl cyclase activity in rat hippocampal membranes with a Kb value of 14 nM. Intracellular recording techniques revealed that AD, (R)-PIA, 5-HT, and 5-carboxyamidotryptamine (5-CT) elicited concentration-dependent hyperpolarization of RMP within the same hippocampal pyramidal cell. The maximal hyperpolarization obtained for the AD or 5-HT analogs was the same for individual pyramidal cells. CPT and spiperone antagonized the hyperpolarization by (R)-PIA and 5-CT, respectively. Saturating concentrations of spiperone failed to antagonize (R)-PIA-mediated responses and CPT did not block responses elicited by 5-HT in either the biochemical or electrophysiological preparations. The combination of saturating concentrations of 5-HT and (R)-PIA evoked nonadditive biochemical responses relative to those observed with (R)-PIA alone. Similarly, electrophysiological experiments conducted under voltage-clamp conditions demonstrated that maximally effective concentrations of AD and 5-CT exhibited nonadditive behavior. Because the amount of outward current elicited when these agonists were coperfused was significantly less than the algebraic sum of the currents evoked individually by these agents, we infer that a population of AD A1 and 5-HT1A receptors activates a common pool of guanine nucleotide-binding proteins. Central administration of pertussis toxin produced similar, dose-dependent reductions in 5-HT- and (R)-PIA-mediated inhibition of forskolin-stimulated adenylyl cyclase activity and hyperpolarization of RMP. In summary, these results demonstrate that AD A1 and 5-HT1A receptors are colcalized on the same hippocampal pyramidal cells and suggest that a common pool of pertussis toxin-sensitive guanine nucleotide-binding proteins couple these receptors to both the reduction of adenylyl cyclase activity and hyperpolarization of RMP.