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Mental Health Research Institute, University of Michigan Ann Arbor, Michigan 48109–0720
Department of Behavioral Neuroscience, University of Pittsburgh Pittsburgh, Pennsylvania 15260
Address requests for reprints to: Dr. James P. Herman, Mental Health Research Institute, 205 Zina Pitcher Place, University of Michigan, Ann Arbor, Michigan 48109–0720.
Abstract
In situ hybridization histochemistry with a probe directed against an intron sequence of the rat arginine vasopressin (AVP) gene was used to demonstrate localization and regulation of AVP heteronuclear RNA in discrete brain regions. Hybridization with an AVP intron I (AVPinl) probe revealed specific hybridization confined to cell nuclei of paraventricular nucleus, supraoptic nucleus (SON), and suprachiasmatic nucleus neurons of the rat hypothalamus. Grain counts revealed that the signal generated by the AVPinl probe represented 1.9% of that derived from an AVP exon C probe (AVPexC) in the SON. Interestingly, in the suprachiasmatic nucleus the proportion of AVPinl to AVP exon C ratio was much higher (12%), suggesting either increased transcription of the AVP gene or changes in posttranscriptional RNA processing. Regulatory experiments revealed that 2.6-fold increases in AVPinl signal could be visualized in the SON as little as 30 min after an acute salt load, a period during which no significant change in cytoplasmic AVP mRNA could be observed. In response to chronic salt loading, both AVP heteronuclear RNA and AVP mRNA were upregulated. These data compared favorably with transcription rate values determined by nuclear run-on assay, suggesting that intronic in situ hybridization affords a relatively reliable method for assessment of rapid changes in gene transcription in individual central nervous system neurons.
FOOTNOTES
This research was supported by NIH Grants NS-08267 (to J.P.H.), BNS-9021307 (to T.G.S.), DA-02265 (to S.J.W.), and MH-42251 (to S.J.W.).
Received for publication June 7, 1991. Revision received July 19, 1991. Accepted for publication July 24, 1991.
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