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Titolo:
IMMUNOLABELING OF THE RAT INTESTINAL-TRACT WITH ANTIBODIES SPECIFIC TO THE LONG FORM OF THE 5-HYDROXYTRYPTAMINE(3) RECEPTOR
Autore:
DOUCET E; HAMON M; EMERIT MB;
Indirizzi:
UNIV PARIS 06,INSERM,U288,91 BLVD HOP F-75634 PARIS 13 FRANCE UNIV PARIS 06,INSERM,U288 F-75634 PARIS 13 FRANCE
Titolo Testata:
Neuroscience
fascicolo: 3, volume: 87, anno: 1998,
pagine: 691 - 707
SICI:
0306-4522(1998)87:3<691:IOTRIW>2.0.ZU;2-E
Fonte:
ISI
Lingua:
ENG
Soggetto:
IN-SITU HYBRIDIZATION; GATED ION-CHANNEL; ENTERIC NERVOUS-SYSTEM; 5-HT3 BINDING-SITES; PIG SMALL-INTESTINE; 2 SPLICE VARIANTS; PHARMACOLOGICAL CHARACTERIZATION; FUNCTIONAL EXPRESSION; H-3 ZACOPRIDE; RECOGNITION SITES;
Keywords:
5-HT3 RECEPTORS; SPLICE VARIANTS; ANTIBODIES; IMMUNOPRECIPITATION; IMMUNOLABELING; INTESTINE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
54
Recensione:
Indirizzi per estratti:
Citazione:
E. Doucet et al., "IMMUNOLABELING OF THE RAT INTESTINAL-TRACT WITH ANTIBODIES SPECIFIC TO THE LONG FORM OF THE 5-HYDROXYTRYPTAMINE(3) RECEPTOR", Neuroscience, 87(3), 1998, pp. 691-707

Abstract

The mouse 5-hydroxytryptamine(3) (5-HT3) type of serotonin receptors is expressed as two forms, 5-HT3R-A(L) and 5-HT3R-A(S), generated by alternative splicing of its primary transcript, that differ by a stretch of six amino acids in the second intracellular loop domain. Because this sis-amino acid region contains a putative phosphorylation site that may be important for the Function and/or regulation of 5HT(3)R-A(L)receptor, specifically, we developed polyclonal antibodies as appropriate tools for studies relevant to this question. Antibodies against a20-amino acid peptide corresponding to the sequence of 5-HT3R-A(L) atthe level of this six-amino acid region were obtained as soon as one month after injection of this synthetic peptide to rabbits. Immunocytochemistry with these antibodies led to a strong positive labelling of plasma membrane, reticulum and Golgi apparatus of COS-7 cells expressing cloned murine 5-HT3R-A(L), whereas COS-7 cells expressing similar levels of 5-HT3R-A(S) exhibited only a very weak labelling. Immunoblotsof fusion proteins combining glutathion-S-transferase and the second cytoplasmic loop of 5-HT3R-A(L) or 5-HT3R-A(S) revealed a c. 20-fold selectivity of the antibodies for the first, long form, as evaluated bydensitometric analysis of enhanced chemiluminescence detection. Similarly, immunoblots of COS-7 cells transfected with cloned 5-MT3 receptors showed that the anti-peptide antibodies detected a band at 53.000 mel. wt only in cells transfected with 5-HT3R-A(L). finder optimal conditions, antibodies immunoprecipitated 52% of 5-HT3R-A(L), but only 11%of 5-HT3R-A(L), solubilized from COS-7 cells transfected with the respective encoding plasmids. In the rat, no immunoautoradiographic labelling by the anti-peptide antibodies could be detected in brain structures which had previously been described to express preferentially a short form of the 5-HT3 receptor. In contrast, a strong immunolabelling was found in the intestinal mucosa: especially in the rat fetus (at the 17th embryonic day). suggesting the possible participation of the 5-HT3R-A(L) isoform in the development of this tissue. These results show that specific antibodies are useful tools for the Visualization of the least abundant 5-HT3 receptor isoform in rat tissue. (C) 1998 IBRO. Published by Elsevier Science Ltd.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 09/04/20 alle ore 07:30:16