Visualisation of non-glycosylated somatostatin receptor two (ngsst2) immunoreactivity in the rat central nervous system

被引:8
作者
Hervieu, G [1 ]
Emson, PC [1 ]
机构
[1] Babraham Inst, Dept Neurobiol, Lab Cognit & Mol Neurosci, Cambridge CB2 4AT, England
来源
MOLECULAR BRAIN RESEARCH | 1998年 / 58卷 / 1-2期
基金
英国医学研究理事会;
关键词
somatostatin; somatostatin receptor 2; immunocytochemistry; cell line; glycosylation;
D O I
10.1016/S0169-328X(98)00120-X
中图分类号
Q189 [神经科学];
学科分类号
071006 ;
摘要
The biological actions of the neuropeptides somatostatin-14 and -28 are receptor-mediated. To date, five G protein-coupled receptors sst(1) to sst(5) have been characterised pharmacologically and their genes have been cloned. In this study, we used an affinity-purified polyclonal antibody (AS-68) raised against a specific N-terminal peptide sequence of sst2 to localise N-terminal sst2-immunoreactive regions in the rat brain and the cervical spinal cord. The specificity of the antiserum was demonstrated by Western and slot blotting experiments using a N-terminal sst2 fusion protein. Further blotting experiments with a sst2((A))-transfected cell line and rat CNS membrane proteins showed that the antibody detected the non-glycosylated and/or non-sialated receptor. A strong signal using an sst2((A))-transfected CHO-K1 cell line was obtained only if the cells had been treated with N-Glycosidase F prior to the immunochemical detection. Two variants of sst2 (sst2((A)) and sst2((B))) have been identified by cloning procedures and gene expression studies in the rodents. They differ in their carboxy-termini: AS-68 would, however, be able to recognise the non-glycosylated form of both these variants. We present here the central nervous system distribution of non-glycosylated sst2-immunoreactivity in the rat using this N-terminal antibody. The sst2 non-glycosylated N-terminal like immunoreactivity was distributed throughout the brain with cells and processes labelled in the cerebral cortex and the basal ganglia (neostriatum, substantia nigra), in the limbic system (hippocampal formation, amygdala), in the diencephalon (epithalamus, thalamus, hypothalamus), the superior colliculus, the periaqueductal grey matter and some of the reticular formation nuclei. The distribution of the non-glycosylated sst2-like immunoreactivity detected here was consistent with that predicted from the localisation of sst2 mRNA and SRIF-ligand binding studies. (C) 1998 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:138 / 155
页数:18
相关论文
共 62 条
[1]   PATTERNS OF EXPRESSION OF SSTR1 AND SSTR2 SOMATOSTATIN RECEPTOR SUBTYPES IN THE HYPOTHALAMUS OF THE ADULT-RAT - RELATIONSHIP TO NEUROENDOCRINE FUNCTION [J].
BEAUDET, A ;
GREENSPUN, D ;
RAELSON, J ;
TANNENBAUM, GS .
NEUROSCIENCE, 1995, 65 (02) :551-561
[2]  
BEAUDET A, 1995, CIBA GEIGY S, P142
[3]   MOLECULAR-BIOLOGY OF SOMATOSTATIN RECEPTORS [J].
BELL, GI ;
REISINE, T .
TRENDS IN NEUROSCIENCES, 1993, 16 (01) :34-38
[4]   GROWTH HORMONE-RELEASING HORMONE-SYNTHESIZING NEURONS ARE A SUBPOPULATION OF SOMATOSTATIN RECEPTOR-LABELED CELLS IN THE RAT ARCUATE NUCLEUS - A COMBINED INSITU HYBRIDIZATION AND RECEPTOR LIGHT-MICROSCOPIC AUTORADIOGRAPHIC STUDY [J].
BERTHERAT, J ;
DOURNAUD, P ;
BEROD, A ;
NORMAND, E ;
BLOCH, B ;
ROSTENE, W ;
KORDON, C ;
EPELBAUM, J .
NEUROENDOCRINOLOGY, 1992, 56 (01) :25-31
[5]   TRANSIENT EXPRESSION OF SOMATOSTATIN RECEPTORS IN THE RAT VISUAL-SYSTEM DURING DEVELOPMENT [J].
BODENANT, C ;
LEROUX, P ;
GONZALEZ, BJ ;
VAUDRY, H .
NEUROSCIENCE, 1991, 41 (2-3) :595-606
[6]   HYPOTHALAMIC POLYPEPTIDE THAT INHIBITS SECRETION OF IMMUNOREACTIVE PITUITARY GROWTH-HORMONE [J].
BRAZEAU, P ;
VALE, W ;
BURGUS, R ;
LING, N ;
BUTCHER, M ;
RIVIER, J ;
GUILLEMIN, R .
SCIENCE, 1973, 179 (4068) :77-79
[7]  
BREDER CD, 1992, J NEUROSCI, V12, P3920
[8]   ENDOCRINE PEPTIDES MOONLIGHTING AS IMMUNE MODULATORS - ROLES FOR SOMATOSTATIN AND GH-RELEASING FACTOR [J].
CAMPBELL, RM ;
SCANES, CG .
JOURNAL OF ENDOCRINOLOGY, 1995, 147 (03) :383-396
[9]  
Dournaud P, 1996, J NEUROSCI, V16, P4468
[10]   COMBINED AUTORADIOGRAPHIC AND IMMUNOHISTOCHEMICAL EVIDENCE FOR AN ASSOCIATION OF SOMATOSTATIN BINDING-SITES WITH GROWTH HORMONE-RELEASING FACTOR-CONTAINING NERVE-CELL BODIES IN THE RAT ARCUATE NUCLEUS [J].
EPELBAUM, J ;
MOYSE, E ;
TANNENBAUM, GS ;
KORDON, C ;
BEAUDET, A .
JOURNAL OF NEUROENDOCRINOLOGY, 1989, 1 (02) :109-115