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. 1987 Jan;382:523–535. doi: 10.1113/jphysiol.1987.sp016382

Neurotensin and acetylcholine evoke common responses in frog oocytes injected with rat brain messenger ribonucleic acid.

C Hirono, I Ito, H Sugiyama
PMCID: PMC1183039  PMID: 2442367

Abstract

1. The intracellular reaction mechanism underlying electrophysiological responses evoked by neurotensin (NT) was studied using Xenopus laevis oocytes injected with poly (A)+ messenger ribonucleic acid (mRNA) isolated from rat brains. 2. A few days after the injection of mRNA, oocytes were found to acquire sensitivity to NT and substance P. 3. Under voltage-clamp conditions (-60 mV), application of NT to mRNA-injected oocytes produced transient and oscillatory inward currents which began after a delay of several tens of seconds. These inward currents were accompanied by an increase in membrane conductance. 4. NT receptors on mRNA-injected oocytes showed essentially the same pharmacological properties as those of native NT receptors. 5. The NT response showed desensitization and was not readily recovered even after extensive washing of cells for more than 30 min. 6. NT response was suppressed when the muscarinic acetylcholine (ACh) response of the same cell, which was also induced by the same mRNA, was desensitized by a large dose of ACh. 7. NT response and ACh response showed many similarities: they were both inhibited by pertussis toxin and intracellular ethyleneglycol-bis-(beta-aminoethylether) N, N'-tetraacetic acid (EGTA), mimicked by intracellularly injected inositol 1, 4, 5-trisphosphate (InsP3), and suppressed when cell response to InsP3 was desensitized by a large dose of InsP3. Reversal-potential analyses indicated that both responses were mediated by an increase in membrane permeability to Cl-. 8. It is concluded that NT responses and muscarinic ACh responses of Xenopus oocytes induced by rat brain mRNA may most likely share a common reaction mechanism. The reaction sequence includes the activation of receptors, activation of inhibitory guanine nucleotide-binding regulatory protein, production of InsP3, intracellular Ca2+ mobilization, and increased membrane permeability to Cl-.

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Selected References

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  1. Amar S., Mazella J., Checler F., Kitabgi P., Vincent J. P. Regulation of cyclic GMP levels by neurotensin in neuroblastoma clone N1E115. Biochem Biophys Res Commun. 1985 May 31;129(1):117–125. doi: 10.1016/0006-291x(85)91411-1. [DOI] [PubMed] [Google Scholar]
  2. Araki K., Tachibana S., Uchiyama M., Nakajima T., Yasuhara T. Isolation and structure of a new active peptide xenopsin on rat stomach strip and some biogenic amines in the skin of Xenopus laevis. Chem Pharm Bull (Tokyo) 1975 Dec;23(12):3132–3140. doi: 10.1248/cpb.23.3132. [DOI] [PubMed] [Google Scholar]
  3. Baldino F., Jr, Davis L. G., Wolfson B. Structure-activity studies with carboxy- and amino-terminal fragments of neurotensin on hypothalamic neurons in vitro. Brain Res. 1985 Sep 9;342(2):266–272. doi: 10.1016/0006-8993(85)91125-4. [DOI] [PubMed] [Google Scholar]
  4. Checler F., Labbé C., Granier C., van Rietschoten J., Kitabgi P., Vincent J. P. [TRP11]-neurotensin and xenopsin discriminate between rat and guinea-pig neurotensin receptors. Life Sci. 1982 Sep 13;31(11):1145–1150. doi: 10.1016/0024-3205(82)90089-3. [DOI] [PubMed] [Google Scholar]
  5. Dascal N., Gillo B., Lass Y. Role of calcium mobilization in mediation of acetylcholine-evoked chloride currents in Xenopus laevis oocytes. J Physiol. 1985 Sep;366:299–313. doi: 10.1113/jphysiol.1985.sp015799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dascal N., Landau E. M., Lass Y. Xenopus oocyte resting potential, muscarinic responses and the role of calcium and guanosine 3',5'-cyclic monophosphate. J Physiol. 1984 Jul;352:551–574. doi: 10.1113/jphysiol.1984.sp015310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gorden P., Carpentier J. L., Cohen S., Orci L. Epidermal growth factor: morphological demonstration of binding, internalization, and lysosomal association in human fibroblasts. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5025–5029. doi: 10.1073/pnas.75.10.5025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gundersen C. B., Miledi R., Parker I. Messenger RNA from human brain induces drug- and voltage-operated channels in Xenopus oocytes. 1984 Mar 29-Apr 4Nature. 308(5958):421–424. doi: 10.1038/308421a0. [DOI] [PubMed] [Google Scholar]
  9. Haga K., Haga T., Ichiyama A., Katada T., Kurose H., Ui M. Functional reconstitution of purified muscarinic receptors and inhibitory guanine nucleotide regulatory protein. Nature. 1985 Aug 22;316(6030):731–733. doi: 10.1038/316731a0. [DOI] [PubMed] [Google Scholar]
  10. Hirono C., Yamagishi S., Ohara R., Hisanaga Y., Nakayama T., Sugiyama H. Characterization of mRNA responsible for induction of functional sodium channels in Xenopus oocytes. Brain Res. 1985 Dec 16;359(1-2):57–64. doi: 10.1016/0006-8993(85)91412-x. [DOI] [PubMed] [Google Scholar]
  11. Houamed K. M., Bilbe G., Smart T. G., Constanti A., Brown D. A., Barnard E. A., Richards B. M. Expression of functional GABA, glycine and glutamate receptors in Xenopus oocytes injected with rat brain mRNA. 1984 Jul 26-Aug 1Nature. 310(5975):318–321. doi: 10.1038/310318a0. [DOI] [PubMed] [Google Scholar]
  12. Kusano K., Miledi R., Stinnakre J. Cholinergic and catecholaminergic receptors in the Xenopus oocyte membrane. J Physiol. 1982 Jul;328:143–170. doi: 10.1113/jphysiol.1982.sp014257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Oron Y., Dascal N., Nadler E., Lupu M. Inositol 1,4,5-trisphosphate mimics muscarinic response in Xenopus oocytes. Nature. 1985 Jan 10;313(5998):141–143. doi: 10.1038/313141a0. [DOI] [PubMed] [Google Scholar]
  14. Parker I., Gundersen C. B., Miledi R. Intracellular Ca2+-dependent and Ca2+-independent responses of rat brain serotonin receptors transplanted to Xenopus oocytes. Neurosci Res. 1985 Aug;2(6):491–496. doi: 10.1016/0168-0102(85)90021-5. [DOI] [PubMed] [Google Scholar]
  15. Pastan I. H., Willingham M. C. Receptor-mediated endocytosis of hormones in cultured cells. Annu Rev Physiol. 1981;43:239–250. doi: 10.1146/annurev.ph.43.030181.001323. [DOI] [PubMed] [Google Scholar]
  16. Quirion R., Rioux F., Regoli D., St-Pierre S. Selective blockade of neurotensin-induced coronary vessel constriction in perfused rat hearts by a neurotensin analogue. Eur J Pharmacol. 1980 Feb 8;61(3):309–312. doi: 10.1016/0014-2999(80)90133-8. [DOI] [PubMed] [Google Scholar]
  17. Rioux F., Quirion R., Regoli D., Leblanc M. A., St-Pierre S. Pharmacological characterization of neurotensin receptors in the rat isolated portal vein using analogues and fragments of neurotensin. Eur J Pharmacol. 1980 Sep 5;66(4):273–279. doi: 10.1016/0014-2999(80)90459-8. [DOI] [PubMed] [Google Scholar]
  18. Sibley D. R., Lefkowitz R. J. Molecular mechanisms of receptor desensitization using the beta-adrenergic receptor-coupled adenylate cyclase system as a model. Nature. 1985 Sep 12;317(6033):124–129. doi: 10.1038/317124a0. [DOI] [PubMed] [Google Scholar]
  19. Sternweis P. C., Robishaw J. D. Isolation of two proteins with high affinity for guanine nucleotides from membranes of bovine brain. J Biol Chem. 1984 Nov 25;259(22):13806–13813. [PubMed] [Google Scholar]
  20. Sugiyama H., Hisanaga Y., Hirono C. Induction of muscarinic cholinergic responsiveness in Xenopus oocytes by mRNA isolated from rat brain. Brain Res. 1985 Jul 15;338(2):346–350. doi: 10.1016/0006-8993(85)90166-0. [DOI] [PubMed] [Google Scholar]
  21. Sumikawa K., Houghton M., Emtage J. S., Richards B. M., Barnard E. A. Active multi-subunit ACh receptor assembled by translation of heterologous mRNA in Xenopus oocytes. Nature. 1981 Aug 27;292(5826):862–864. doi: 10.1038/292862a0. [DOI] [PubMed] [Google Scholar]
  22. Sumikawa K., Parker I., Miledi R. Partial purification and functional expression of brain mRNAs coding for neurotransmitter receptors and voltage-operated channels. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7994–7998. doi: 10.1073/pnas.81.24.7994. [DOI] [PMC free article] [PubMed] [Google Scholar]

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