Oxytocin and vasopressin gene expression and RNA splicing patterns in the rat supraoptic nucleus

Molecular Neuroscience Section, Laboratory of Neurochemistry, National Institute of Neurological Disorders and Stroke; National Institutes of Health; Bethesda, Maryland In this study, we test the hypothesis that there are differential splicing patterns between the expressed oxytocin (OT) and vasopre...

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Bibliographic Details
Published in:Physiological genomics 2008-11, Vol.35 (3), p.231-242
Main Authors: Yue, Chunmei, Ponzio, Todd A, Fields, Raymond L, Gainer, Harold
Format: Article
Language:English
Subjects:
Animals
Female
Gene Expression Profiling
In Situ Hybridization
Male
Oxytocin - genetics
Rats
Rats, Sprague-Dawley
Reverse Transcriptase Polymerase Chain Reaction
RNA Splicing - genetics
Supraoptic Nucleus - metabolism
Vasopressins - genetics
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Summary:Molecular Neuroscience Section, Laboratory of Neurochemistry, National Institute of Neurological Disorders and Stroke; National Institutes of Health; Bethesda, Maryland In this study, we test the hypothesis that there are differential splicing patterns between the expressed oxytocin (OT) and vasopressin (VP) genes in the rat supraoptic nucleus (SON). We quantify the low abundance, intron-containing heteronuclear RNAs (hnRNAs) and the higher abundance mRNAs in the SON using two-step, quantitative SYBR Green real-time reverse transcription (RT)-PCR and external standard curves constructed using synthetic 90 nt sense-strand oligonucleotides. The levels of OT and VP mRNA in the SON were found to be similar, 10 8 copies/SON pair, whereas the copy numbers of VP hnRNAs containing intron 1 or 2 and the OT hnRNA containing intron 1 are much lower, i.e., 10 2 –10 3 copies/rat SON pair. However, the estimated copy number of the intron 2-containing OT hnRNA is much larger, 10 6 copies/SON pair. The relative distributions of all the OT and VP RNA species were invariant and independent of the physiological status of the rats (e.g., osmotically stimulated or lactating rats). Using intron-specific riboprobes against hnRNAs, we demonstrate by fluorescence in situ hybridization strong signals of OT hnRNA containing intron 2 predominantly in the cytoplasm, in contrast to the localization of the VP hnRNA found only in the nuclei. Taken together, these data support the view that the splicing patterns between OT and VP gene transcripts are different and show that there is a selective cytoplasmic retention of OT intron 2. real-time polymerase chain reaction; intron retention; hypothalamus
ISSN:1094-8341
1531-2267
DOI:10.1152/physiolgenomics.90218.2008