Measurement of nitric oxide-related enzymes in the brain by in situ hybridization

Détails

ID Serval
serval:BIB_8C0D39178ED6
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Measurement of nitric oxide-related enzymes in the brain by in situ hybridization
Périodique
Methods in Molecular Biology
Auteur(s)
Braissant O.
ISSN
1064-3745 (Print)
Statut éditorial
Publié
Date de publication
2004
Volume
279
Pages
113-124
Notes
Journal Article Research Support, Non-U.S. Gov't
Résumé
Functional genomic increasingly requires the cell-specific localization of gene expression. The histological analysis of mRNA expression is performed by in situ hybridization (ISH), whereas proteins are detected by immunohistochemistry. ISH allows the rapid detection of any transcript with the same protocol provided that its cDNA is known. This is an advantage compared to immunohistochemistry, which necessitates the raise of specific antibodies and the test of different procedures depending on the antigens to detect. However, ISH is subject to multiple technical difficulties. We have developed a nonradioactive ISH protocol allowing the simultaneous coupling of immunohistochemistry to follow gene expression at the protein level on the same section as the mRNA detection. This chapter describes the neuronal nitric oxide synthase (nNOS), argininosuccinate synthetase (AS), and argininosuccinate lyase (AL) mRNA expression in the rat brain to illustrate the methods allowing the nonradioactive localization of transcripts by ISH, coupled to immunohistochemistry to identify the specific cell types of central nervous system (CNS) on the same section.
Mots-clé
Animals Argininosuccinate Lyase/*metabolism Argininosuccinate Synthase/*metabolism Brain/anatomy & histology/*enzymology Immunohistochemistry/methods In Situ Hybridization/methods Nitric Oxide/metabolism Nitric Oxide Synthase/*metabolism Nitric Oxide Synthase Type I RNA, Messenger/*metabolism Rats
Pubmed
Création de la notice
25/01/2008 15:21
Dernière modification de la notice
03/03/2018 19:12
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