Spatiotemporal profiles of arginine vasopressin transcription in cultured suprachiasmatic nucleus.

摘要: Arginine vasopressin (AVP), a major neuropeptide in the suprachiasmatic nucleus (SCN), is postulated to mediate output of circadian oscillation. Mice carrying reporter gene AVP transcription (AVP(ELuc)) were produced by knocking-in cDNA Emerald-luciferase (ELuc) translational initiation site. Homozygous mice did not survive beyond postnatal day 7. Using heterozygous (AVP(ELuc/+)) mice, bioluminescence system was developed that enabled monitor through AVP-ELuc measurement real time for more than 10 cycles cultured brain slice. AVP(ELuc/+) showed behaviour rhythms and light responsiveness indistinguishable from those wild-type. Robust detected SCN slice at single cell as well tissue levels. The rhythm whole stable, with peak mid-light phase light-dark cycle, while variable. By comparison, rhythmicity paraventricular supraoptic hypothalamus unstable damped rapidly. Spatiotemporal profiles expression pixel level revealed significant entire area AVP-positive cells SCN, least two clusters different oscillations. Contour analysis intensity cell-like region demonstrated radiation almost identical size. This newly useful tool study rhythms.