Experimental approaches for the study of oxytocin and vasopressin gene expression in the central nervous system.
Intron-specific probes measure heteronuclear RNA (hnRNA) levels and thus approximate the transcription rates of genes, in part because of the rapid turnover of this intermediate form of RNA in the cell nucleus. Previously, we used oxytocin (Oxt)- and vasopressin (Avp)- intron-specific riboprobes to measure changes in Oxt and Avp hnRNA levels in the supraoptic nucleus (SON) by quantitative in situ hybridization (ISH) after various classical physiological perturbations, including acute and chronic salt loading, and lactation. In the present experiments, we used a novel experimental model to study the neurotransmitter regulation of Oxt and Avp gene expression in the rat SON in vivo. Bilateral cannulae connected via tubing to Alzet osmotic mini-pumps were positioned over the SON. In every experiment, one SON was infused with PBS and served as the control SON in each animal, and the contralateral SON received infusions of various neurotransmitter agonists and antagonists. Using this approach, we found that Avp but not Oxt gene expression increased after acute (2-5h) combined excitatory amino acid agonist and GABA antagonist treatment, similar to what we found after an acute hyperosmotic stimulus. Since both OXT and AVP are known to be comparably and robustly secreted in response to acute osmotic stimuli in vivo and glutamate agonists in vitro, our results indicate a dissociation between OXT secretion and Oxt gene transcription in vivo.
Scordalakes
Elka M
EM
Laboratory of Neurochemistry, National Institute of Neurological Disorder and Stroke, National Institutes of Health, Bethesda, MD, USA.
Yue
Chunmei
C
Gainer
Harold
H
D000818:Animals; D002490:Central Nervous System / Q000502:physiology*; D017931:DNA Primers; D017403:In Situ Hybridization; D007438:Introns; D008297:Male; D010121:Oxytocin / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012332:RNA, Heterogeneous Nuclear / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D013495:Supraoptic Nucleus / Q000502:physiology*; D014158:Transcription, Genetic; D014667:Vasopressins / Q000235:genetics* / Q000378:metabolism
D017931:DNA Primers; D012332:RNA, Heterogeneous Nuclear; D014667:Vasopressins; D010121:Oxytocin