Efficient coupling of 5-HT1a receptors to the phospholipase C pathway in Xenopus oocytes.
To investigate the receptor-channel coupling pathway, the coding region of the 5-HT1a receptor was subcloned into two plasmid vectors pSP64(polyA+) and pSP64T. Compared to the original 5-HT1a receptor construct G-21, both new constructs increased greatly the expression of functional 5-HT1a receptors in Xenopus oocytes, which developed large inward current responses to 5-HT. These responses were dose-dependent (EC50 approximately 150 nM), and could be elicited also by 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT). The 5-HT1a receptor mediated current had an oscillatory time course, and a reversal potential close to the equilibrium potential for Cl- (ca. -25 mV). Moreover, during and for some minutes following the application of 5-HT, these oocytes acquired the property of generating a transient inward current when their membrane was hyperpolarized. These features are characteristic of responses mediated by other receptors (e.g. muscarinic, angiotensin, serum receptors, etc.) that are known to couple to the endogenous PLC/PI second messenger pathway in Xenopus oocytes. In particular, the 5-HT1a receptor mediated current was very similar to the current induced by 5-HT-stimulation of heterogenic 5-HT2c receptors. Our results show further that the 5-HT1a receptor couples to the endogenous PLC/PI pathway much less efficiently than the 5-HT2c receptor. These results demonstrate clearly that the human 5-HT1a receptor can couple efficiently to the Xenopus oocyte endogenous PLC/PI pathway, and provide additional evidence for cell-specific signal transduction.
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Department of Psychobiology, University of California, Irvine 92697-4550, USA.
Panicker
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Miledi
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D017371:8-Hydroxy-2-(di-n-propylamino)tetralin / Q000494:pharmacology*; D000818:Animals; D003001:Cloning, Molecular; D004298:Dopamine / Q000494:pharmacology; D005260:Female; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects; D009638:Norepinephrine / Q000494:pharmacology; D009865:Oocytes / Q000502:physiology*; D010716:Phosphatidylinositols / Q000378:metabolism; D014176:Protein Biosynthesis; D051381:Rats; D044405:Receptor, Serotonin, 5-HT2C; D011985:Receptors, Serotonin / Q000096:biosynthesis / Q000502:physiology*; D044263:Receptors, Serotonin, 5-HT1; D011994:Recombinant Proteins / Q000096:biosynthesis; D015290:Second Messenger Systems; D012701:Serotonin / Q000494:pharmacology*; D017366:Serotonin Receptor Agonists / Q000494:pharmacology; D014158:Transcription, Genetic; D010738:Type C Phospholipases / Q000378:metabolism*; D014981:Xenopus
D010716:Phosphatidylinositols; D044405:Receptor, Serotonin, 5-HT2C; D011985:Receptors, Serotonin; D044263:Receptors, Serotonin, 5-HT1; D011994:Recombinant Proteins; D017366:Serotonin Receptor Agonists; D012701:Serotonin; D017371:8-Hydroxy-2-(di-n-propylamino)tetralin; D010738:Type C Phospholipases; D004298:Dopamine; D009638:Norepinephrine