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Titolo:
CORTICOTROPIN-RELEASING HORMONE-RECEPTOR TYPE-1 - GENERATION AND CHARACTERIZATION OF POLYCLONAL ANTIPEPTIDE ANTIBODIES AND THEIR LOCALIZATION IN PITUITARY-CELLS AND CORTICAL-NEURONS IN-VITRO
Autore:
CASTRO MG; MORRISON E; PERONE MJ; BROWN OA; MURRAY CA; AHMED I; PERKINS AV; EUROPEFINNER G; LOWENSTEIN PR; LINTON EA;
Indirizzi:
UNIV MANCHESTER,DEPT MED,MOLEC MED UNIT,STOPFORD BLDG,RM 1-302,OXFORDRD MANCHESTER M13 9PT LANCS ENGLAND UNIV OXFORD,JOHN RADCLIFFE MATERN HOSP,NUFFIELD DEPT OBSTET & GYNAECOL OXFORD OX3 9UD ENGLAND
Titolo Testata:
Journal of neuroendocrinology
fascicolo: 7, volume: 8, anno: 1996,
pagine: 521 - 531
SICI:
0953-8194(1996)8:7<521:CHT-GA>2.0.ZU;2-N
Fonte:
ISI
Lingua:
ENG
Soggetto:
ANTERIOR-PITUITARY; HUMAN MYOMETRIUM; BINDING PROTEIN; PLASMA-MEMBRANE; RAT PITUITARY; CRF RECEPTORS; TUMOR-CELLS; IN-VITRO; IDENTIFICATION; BRAIN;
Keywords:
CRH-RECEPTOR 1 ANTISERUM; ANTERIOR PITUITARY; ATT20; NEURONS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Science Citation Index Expanded
Citazioni:
42
Recensione:
Indirizzi per estratti:
Citazione:
M.G. Castro et al., "CORTICOTROPIN-RELEASING HORMONE-RECEPTOR TYPE-1 - GENERATION AND CHARACTERIZATION OF POLYCLONAL ANTIPEPTIDE ANTIBODIES AND THEIR LOCALIZATION IN PITUITARY-CELLS AND CORTICAL-NEURONS IN-VITRO", Journal of neuroendocrinology, 8(7), 1996, pp. 521-531

Abstract

Corticotrophin-releasing hormone (CRH) is a 41 amino acid neuropeptide which plays a major role in regulating the endocrine response to stress. CRH acts by first binding to specific receptors on the plasma membrane of target cells. A CRH receptor from a human corticotroph adenoma and rat brain has recently been cloned (CRH-R1). In this paper, we have chosen three different peptide sequences within the CRH-R1 molecule which bear no similarity to other members of this receptor subfamily(or indeed any known protein) and which are likely to be exposed on the surface of the native protein, for antibody production. Some of these fragments produced antipeptide antibodies of good titre which cross-reacted with the CRH-R1 receptor expressed in transiently transfectedCOS-7 cells and in tissue extracts from rat cerebellum, cortex, pituitary gland and human myometrium, both in Western blots and in liquid-phase radioimmunoassay. We used immunofluorescence techniques to localize the CRH receptor in transiently transfected COS-7 cells, primary cultures of rat anterior pituitary (AP) cells, the corticotroph-tumour cells AtT20 D16-16 and cortical neurons in primary culture. Our resultsindicate IR-CRH-R1 receptors have a punctate distribution on the plasma membrane of AP cells and AtT20 D16-16 cells. Whilst in AP cells their appearance is a fine punctate pattern, in AtT20 cells, they appear as large patches which could account for receptor clusters. Within primary cortical neurons, their distribution does not appear to be polarized. Our results suggest that distribution of CRH-R1 receptors within the different cell-types investigated depends not only on the amino acid sequence but also on cellular factors.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 28/11/20 alle ore 01:01:36