Premium
Guinea Pig Histamine H 1 Receptor. I. Gene Cloning, Characterization, and Tissue Expression Revealed by In Situ Hybridization
Author(s) -
Traiffort E.,
Leurs R.,
Arrang J. M.,
TardivelLacombe J.,
Diaz J.,
Schwartz J.C.,
Ruat M.
Publication year - 1994
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1994.62020507.x
Subject(s) - microbiology and biotechnology , biology , northern blot , 5 ht5a receptor , in situ hybridization , receptor , gabbr1 , complementary dna , southern blot , chinese hamster ovary cell , gene , gene expression , biochemistry , protease activated receptor 2
An intronless DNA encoding the guinea pig H 1 receptor was cloned from a genomic library using probes derived from the bovine H 1 receptor. It encodes a protein of 488 amino acids with a calculated molecular mass of 55,619 daltons compared with a size of 56–68 kDa for the photoaffinity‐labeled receptor as determined by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis analysis. The protein displays a 66% homology with the bovine receptor. Stable expression of the H 1 receptor, characterized by the appearance of [ 3 H]mepyramine binding sites with a pharmacology similar to that of the native H 1 receptor, was obtained following transfection of Chinese hamster ovary cells. Southern blot analysis, using a variety of restriction enzymes, did not provide any evidence of multiple H 1 isoreceptors. Northern blot analysis of a variety of guinea pig peripheral or cerebral tissues identified, in most cases, a single transcript of 3.3 kb, but also, in some tissues, a second transcript of 3.7 kb, possibly generated by the use of different promoter or polyadenylation sites or corresponding to a transcript from a distinct gene. In situ hybridization studies showed the highly contrasted cerebral expression of H 1 ‐receptor gene transcripts, which was compared with autoradiographic receptor localization. This allowed the identification of some major cell populations expressing the H 1 receptor, e.g., Purkinje cells in cerebellum or pyramidal cells in the hippocampal complex.