A hVIPR transgene as a novel tool for the analysis of circadian function in the mouse suprachiasmatic nucleus
A hVIPR transgene as a novel tool for the analysis of circadian function in the mouse suprachiasmatic nucleus
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DOI：
10.1046/j.1460-9568.2003.02487.x
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发表时间：
2003-02-01
影响因子：
3.4
通讯作者：
Hastings, MH
中科院分区：
文献类型：
--
作者：
King, VM;Chahad-Ehlers, S;Hastings, MH
携带编码人VPAC 2受体的新型转基因的小鼠（hVIPR ; Shen et al.（2000）PNAS，97，11575-11580）用于研究下丘脑视交叉上核（SCN）中的昼夜节律功能。通过β-半乳糖苷酶（β-GAL）标签检测到的表达hVPAC 2 R的神经元在SCN内具有独特的分布，与neurophysin（NP）神经元的分布紧密匹配，并延伸到肽组氨酸异亮氨酸（PHI）细胞的区域中。与NP和PHI细胞一样，表达hVPAC 2 R的神经元本质上是昼夜节律的，如mPERIOD（mPER）蛋白的同步节律表达所揭示的。不表达PHI、NP或hVPAC 2 R的SCN细胞群体表现出与SCN的其余部分反相的昼夜PER表达。夜间光照诱导mPER 1在腹侧SCN和mPER 2广泛的整个核。在hVPAC 2 R细胞中诱导核mPER 2证实了它们的光响应性。在建立了它们的昼夜节律特性之后，我们测试了表达hVIPR转基因的SCN神经元作为SCN组织移植物的功能和解剖学明确标记物的效用。来自hVIPR转基因幼仔的产前SCN组织移植到成年CD 1小鼠中存活，并且表达β-GAL、PER和PHI。在一系列的研究中，hVIPR转基因SCN移植物恢复了72例缺血性SCN损伤受者中的17例（23.6%）的昼夜活动节律。通过在杂合Clock突变体背景上使用杂合hVIPR转基因移植物，我们证实了恢复的活动节律是由供体组织赋予的。我们的结论是，hVIPR转基因是一个强大的和灵活的工具，在小鼠SCN的昼夜节律功能的检查。
A mouse bearing a novel transgene encoding the human VPAC2 receptor (hVIPR ; Shen et al . (2000) PNAS , 97, 11575-11580) was used to investigate circadian function in the hypothalamic suprachiasmatic nuclei (SCN). Neurons expressing hVPAC2R, detected by a beta-galactosidase (beta-GAL) tag, have a distinct distribution within the SCN, closely matching that of neurophysin (NP) neurons and extending into the region of peptide histidine isoleucine (PHI) cells. In common with NP and PHI cells, neurons expressing hVPAC2R are circadian in nature, as revealed by synchronous rhythmic expression of mPERIOD (mPER) proteins. A population of SCN cells not expressing PHI, NP or hVPAC2R exhibited circadian PER expression antiphasic with the rest of the SCN. Nocturnal light exposure induced mPER1 in the ventral SCN and mPER2 widely across the nucleus. Induction of nuclear mPER2 in hVPAC2R cells confirmed their photic responsiveness. Having established their circadian properties, we tested the utility of SCN neurons expressing the hVIPR transgene as functionally and anatomically explicit markers for SCN tissue grafts. Prenatal SCN tissue from hVIPR transgenic pups survived transplantation into adult CD1 mice, and expressed beta-GAL, PER and PHI. Over a series of studies, hVIPR transgenic SCN grafts restored circadian activity rhythms to 17 of 72 arrhythmic SCN lesioned recipients (23.6%). By using heterozygous hVIPR transgenic grafts on a heterozygous Clock mutant background we confirmed that restored activity rhythms were conferred by the donor tissue. We conclude that the hVIPR transgene is a powerful and flexible tool for examination of circadian function in the mouse SCN.