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Involvement of a GTP-binding protein in mediation of serotonin and acetylcholine responses in Xenopus oocytes injected with rat brain messenger RNA

Dascal, Nathan and Ifune, Catherine and Hopkins, Rosemary and Snutch, Terry P. and Lübbert, Hermann and Davidson, Norman and Simon, Melvin I. and Lester, Henry A. (1986) Involvement of a GTP-binding protein in mediation of serotonin and acetylcholine responses in Xenopus oocytes injected with rat brain messenger RNA. Molecular Brain Research, 1 (3). pp. 201-209. ISSN 0169-328X. https://resolver.caltech.edu/CaltechAUTHORS:20201012-152225953

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Abstract

Injection of poly(A)⁺ RNA from rat brain into Xenopus oocytes caused the appearance of Cl currents in response to serotonin (5-HT) and acetylcholine (ACh). Both neurotransmitters evoked two-component currents similar in their time course to the oocyte's endogenous cholinergic muscarinic response, which was shown in previous studies to be mediated by IP₃ synthesis leading to Ca release from intracellular stores. The responses to ACh and 5-HT exhibited self- and cross-desensitization, i.e., application of either ACh or 5-HT inhibited the subsequent response to either one of the two transmitters. Intracellular injection of guanosine 5′-O-(3-thiotriphosphate) (GTP-γ-S) mimicked the 5-HT and ACh response, and also completely suppressed the response to the subsequent application of either ACh or 5-HT. Treatment of the oocytes with pertussis toxin (PTX) caused a 50% attenuation of ACh and 5-HT responses. In the membranes of both control and mRNA-injected oocytes, PTX catalyzed the ADP-ribosylation of a single M_r = ∼40,000 protein. Injection of the purified βγ-subunits of transducin enhanced the 5-HT response. The 5-HT and GTP-γ-S responses were inhibited by intracellular injection of the Ca²⁺ chelator, EGTA, as previously shown for the ACh response. These data suggest that ACh and 5-HT receptors, synthesized in the oocytes on the template of brain mRNA, act through a common pathway that involves (a) a guanine nucleotide binding protein and (b) IP₃ production leading to Ca mobilization.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/0169-328x(86)90026-4DOIArticle
ORCID:
AuthorORCID
Lester, Henry A.0000-0002-5470-5255
Additional Information:© 1986 Published by Elsevier. Accepted 9 September 1986. This work was supported by NIH Grants GM-29836 to H.A.L., GM-10991 to N.D., and GM-34236 to M.S. N.D. was a recipient of Bantrell and Fulbright Postdoctoral Fellowships. H.L. was a fellow of the Deutsche Forschungsgemeinschaft. T.S. was a postdoctoral fellow of the American Heart Association/Greater Los Angeles Affiliate. The authors are grateful to Ms. Moira Fearey for excellent technical assistance.
Funders:
Funding AgencyGrant Number
NIHGM-29836
NIHGM-10991
NIHGM-34236
Natural Sciences and Engineering Research Council of Canada (NSERC)UNSPECIFIED
Fulbright FoundationUNSPECIFIED
Deutsche Forschungsgemeinschaft (DFG)UNSPECIFIED
American Heart Association, Greater Los Angeles AffiliateUNSPECIFIED
Subject Keywords:Inositol trisphosphate; Chloride channel; Calcium
Issue or Number:3
Record Number:CaltechAUTHORS:20201012-152225953
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20201012-152225953
Official Citation:Nathan Dascal, Catherine Ifune, Rosemary Hopkins, Terry P. Snutch, Hermann Lübbert, Norman Davidson, Melvin I. Simon, Henry A. Lester, Involvement of a GTP-binding protein in mediation of serotonin and acetylcholine responses in Xenopus oocytes injected with rat brain messenger RNA, Molecular Brain Research, Volume 1, Issue 3, 1986, Pages 201-209, ISSN 0169-328X, https://doi.org/10.1016/0169-328X(86)90026-4.
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:105992
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:13 Oct 2020 14:17
Last Modified:13 Oct 2020 14:17

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