open access

https://hdl.handle.net/2115/63362

Spatiotemporal profiles of arginine vasopressin transcription in cultured suprachiasmatic nucleus

Spatiotemporal profiles of AVP transcription in the SCN

英語

SCN

2015-11

Wiley-Blackwell

European journal of neuroscience

42(9)

2678-2689

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

学術雑誌論文

Accepted Manuscript

This is the peer reviewed version of the following article: Yoshikawa, T., Nakajima, Y., Yamada, Y., Enoki, R., Watanabe, K., Yamazaki, M., Sakimura, K., Honma, S., Honma, K.-i. (2015), Spatiotemporal profiles of arginine vasopressin transcription in cultured suprachiasmatic nucleus. European Journal of Neuroscience, 42: 2678–2689., which has been published in final form at http://dx.doi.org/10.1111/ejn.13061. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.

0953-816X

AA10672270