Investigation on the biological limit cycles by using transgenic mice.

SCN内的双钟系统及喷射滞后的原因我们发现Per 1基因的诱导局限于SCN的腹外侧区，夜间短暂光照后，大鼠的视交叉上核（SCN）腹外侧区出现高水平的Per 1 mRNA的诱导。结果表明，在SCN中存在两种类型的时钟：光响应和光不响应。为了清楚地显示SCN中的双振子系统，我们利用了大的快速亮暗循环移位。LD周期延迟10小时后，我们发现SCN内部计时系统的解离; Per 1在腹外侧区的表达显示出快速的大位移，而背内侧区的影响要小得多。接下来两个区域的同化过程相当缓慢，需要几天时间。这些结果表明，在SCN中存在潜在的两个时钟，它们可以通过一定的扰动而分离。这些发现还表明， ...更多信息 我们将果蝇timeless（tim）启动子驱动的mPer 1和mPer 2基因导入果蝇，研究了果蝇Per基因的功能保守性。行为测定显示，两种mPer构建体都恢复了<01>由于缺乏内源性Per蛋白（PER）而在其他方面是嗜热的Per果蝇中的节律。本研究证明mPer 1和mPer 2都具有生物钟的功能，为不同的per基因的功能分析提供了依据。将这些cDNA对应于用FLAG标记的Per 1、Per 2和Per 3的区域亚克隆到表达载体中。我们将这些表达载体转染到细胞系中，并获得了这些蛋白的足够表达。现在我们正试图建立携带这些Per基因的转基因小鼠。少

Two-clock system in the SCN and cause of jet lagWe found the restriction of Per1 gene induction to the ventrolateral region of the SCN.After brief light exposure in the night, rats showed high level of Per1 mRNA induction in the ventrolateral region of the suprachiasmatic nucleus (SCN). The results suggested that there are two types of clocks in the SCN ; light responsive and light unresponsive. To show the two-oscillator system in the SCN clearly, we utilized a large rapid light-dark cycle shift. After the 10 hours delay of LD cycle, we found the dissociation of the internal timing system in the SCN ; the Per1 expression in the ventrolateral region showed a rapid large shift while the dorsomedial regions were far less affected. The following resynchronization process of the two regions was rather slow, requiring several days. These findings suggest that there are potential two clocks in the SCN, which could be dissociated by a certain perturbation. These findings also suggest that jet … More lag is caused by delayed shift of the clocks in the dorsomedial region of the SCN.Investigation on the transgenic flies carrying mouse Per genesWe investigated the functional conservation of per by introducing the mouse mPer1 and mPer2 genes, driven by the Drosophila timeless (tim) promoter, into Drosophila melanogaster. Behavioral assays showed that both mPer constructs restored rhythms in per^<01>flies that are otherwise arrhythmic due to a lack of endogenous per protein (PER). This study demonstrates that both mPer1 and mPer2 can function as clock components, and has implications for functional analysis of the different per genes.Analysis on the transgenic mouse carrying mPer1 genesWe obtained the cDNAs containing coding region and promoter regions of mPer1, Per2, and Per3. These cDNAs corresponding regions of Per1, Per2 and Per3 tagged with FLAG are subcloned into expression vectors. We transfected these expression vectors into cell lines and obtained enough expression of these proteins. Now we are trying to establish the transgenic mice carrying these Per genes. Less