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In situ hybridization of putative somatostatin mRNA within hypothalamus of the rat using synthetic oligonucleotide probes
Author(s) -
Arentzen Rene,
Baldino Frank,
Davis Leonard G.,
Higgins Gerald A.,
Lin Yuan,
Manning Robert W.,
Wolfson Betty
Publication year - 1985
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.240270410
Subject(s) - in situ hybridization , hypothalamus , somatostatin , messenger rna , microbiology and biotechnology , complementary dna , biology , oligonucleotide , endocrinology , dna , biochemistry , gene
Abstract The distribution of mRNA with high sequence homology to somatostatin mRNA within the periventricular hypothalamus of rat was assessed using in situ hybridization techniques with synthetic oligodeoxyribonucleotide probes, complementary to the 3′ coding region of rat somatostatin mRNA. The probes (22‐ and 24‐mers) were 5′‐end labeled using T4 polynucleotide kinase and γ‐ 32 P‐ATP. They were used either individually or after ligation with T4 DNA ligase to form a 46‐mer. Serial tissue sections ( < 10 μm) were taken from the level of the preoptic/anterior hypothalamus through the paraventricular hypothalamus. In situ hybridizations were conducted at room temperature in hybridization buffer. Neurons immuno‐reactive with antiserum raised against somatostatin were identified in alternate sections using standard immunocytochemical procedures. The anatomical location of the hybridization signal was determined by autoradiography. Our results show that the peri‐ and paraventricular hypothalamus is rich in transcripts putatively coding for somatostatin and that these transcripts are co‐distributed with neurons immunoreactive with antisomatostatin immunoglobulin.