2012 Fiscal Year Final Research Report
Real-time monitoring of GFP-expression in the rat hypothalamus via optical fiber
| Project/Area Number |
23659125
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Environmental physiology (including Physical medicine and Nutritional physiology)
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| Research Institution | Nippon Medical School |
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Principal Investigator |
IIJIMA Norio 日本医科大学, 医学部, 准教授 (00285248)
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| Project Period (FY) |
2011 – 2012
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| Keywords | 遺伝子発現リアルタイムモニタリング / 脳深部 / 光ファイバー / GFP / レーザー / 無麻酔・非拘束 / バソップレシン |
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| Research Abstract |
In vivo and real-time monitoring of gene expression in the deep structure of the brain could bring beneficial information for understanding neuroendocrine and autonomic nervous system. We developed the system which could monitor green fluorescent protein (GFP)-expression in the hypothalamus via optical fiber. One terminal of optical fiber was connected to blue semiconductor laser oscillators/ green fluorescent detector. The other terminal of it was inserted into the hypothalamus and fixed with thecranium. This system enables real-time monitoring of GFP-expression with awake and unrestrained rat. However optical fiber is vulnerable to twist caused by rat’s free moving. Therefore, we also developed the cage which turns the floor in response to turning of rat’s head for relief of twist-stress of optical fiber between laser oscillator / fluorescent detector and the rat’s head. We applied this system to the arginine vasopressin (AVP)-eGFP transgenic rats in which GFP was expressed in the paraventricular nucleus (PVN) and the supraoptic nucleus (SON) in the hypothalamus. Those rats show increase of GFP-expression in the PVN and the SON under salt-loading condition (Ueta et al. Endocrinology, 2005). Using this system, we monitored the change of AVP-expression in the hypothalamus via GFP-expression under dehydration and rehydration for one week. After monitoring via optical fiber, the brain were fixed and cut into slices for observation of the position of fiber-insertion. We are trying to monitor GFP-intensity in the PVN and the SON and that around the GFP-fibers in the hypothalamus. In this study, we show the new monitoring tool ofgene expression in the deep structure of brain.
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Research Products
(2 results)
