Vasopressin mRNA in situ hybridization: localization and regulation studied with oligonucleotide cDNA probes in normal and Brattleboro rat hypothalamus.
Hybridizable vasopressin mRNA may be quantitatively localized in situ in sections from rat hypothalamus. Radiolabeled oligonucleotide cDNA probes, synthesized by chemical and enzymatic means, provide strong hybridization in zones known to contain vasopressin cell bodies. Multiple single-stranded 32P-, 35S-, or 3H-labeled oligonucleotides demonstrate localized hybridization that increases as probes are lengthened from 8 to 75 bases. Competition studies, RNase experiments, anatomic localization, and use of multiple probes all support hybridization specificity. An approximate doubling of hybridizable mRNA in both supraoptic and paraventricular nuclei can be detected with dehydration of the animals. Hybridizable mRNA densities are virtually normal in hypothalamic nuclei of Brattleboro rats given free access to water. These methods can provide insight into regional mRNA dynamics and may reflect functional activity of peptidergic neurons.
D000818:Animals; D001345:Autoradiography; D004247:DNA / Q000378:metabolism*; D007031:Hypothalamus / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides; D010761:Phosphorus Radioisotopes; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D011910:Rats, Brattleboro; D011919:Rats, Inbred Strains; D013045:Species Specificity; D013462:Sulfur Radioisotopes; D014316:Tritium; D014667:Vasopressins / Q000235:genetics*
D009841:Oligonucleotides; D010761:Phosphorus Radioisotopes; D012333:RNA, Messenger; D013462:Sulfur Radioisotopes; D014316:Tritium; D014667:Vasopressins; D004247:DNA