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Quantitation of Vasopressin mRNA and Oxytocin mRNA in Hypothalamic Nuclei by Solution Hybridization Assays
Author(s) -
Burbach J. Peter H.,
Tol Hubert H. M.,
Bakkus Marleen H. C.,
Schmale Hartwig,
Ivell Richard
Publication year - 1986
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 - 10.1111/j.1471-4159.1986.tb13093.x
Subject(s) - messenger rna , supraoptic nucleus , vasopressin , oxytocin , in situ hybridization , microbiology and biotechnology , hypothalamus , biology , rna , precursor mrna , neuropeptide , gene expression , chemistry , endocrinology , medicine , biochemistry , gene , receptor , rna splicing
Concentrations of vasopressin (VP) precursor and oxytocin (OT) precursor mRNA were measured in magno‐cellular cell groups of the rat hypothalamus by newly developed solution hybridization assays. The assays employed single‐stranded 35 S‐labeled VP‐specific and OT‐specific DNA probes that were prepared by primer extension on re‐combinant M13 DNA templates. Solution hybridization assays were standardized by known amounts of cloned DNA. The detection limit was <1 pg DNA equivalent of the respective mRNA. In total RNA preparations of microdis‐sected supraoptic nucleus (SON) mean (SEM) basal levels of 1.370.18pgVP mRNA and 1.950.I4pgOT mRNA were measured. RNA of the microdissected paraventricular nucleus (PVN) contained 0.35 ± 0.02 pg VP mRNA and 1.77 ± 0.15 pg OT mRNA. Elevation of plasma osmolality induced by drinking of 2% saline for 25 days resulted in a 1.85‐fold increase in VP mRNA levels of the SON and a 1.6‐fold increase in VP mRNA levels of the PVN. The solution hybridization assays are suitable tools to study the regulation of VP and OT mRNAs in magnocellular neurons of the brain.