权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Function of Clock Genes : Examination in Cultured Suprachiasmatic Nucleus

时钟基因的功能：培养的视交叉上核的检查

基本信息

批准号：
11670057
负责人：
HONMA Sato
金额：
$ 2.5万
依托单位：
HOKKAIDO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

Clock gene functions were examined in single suprachiasmatic neurons by culturing suprachiasmatic nucleus (SCN) of new born rats or mice on an multi electrode dish. Spontaneous firing of single neurons was monitored continuously for up to several weeks and circadian rhythms in firing rate were analyzed.1.BMAL1 gene function in the circadian firing rhythms of individual SCN neuronsAntisense oligonucleotide was applied on SCN neurons showing robust circadian firing rhythms, and effects on rhythm parameters, such as phase, period, and amplitude, were examined at the level of single SCN neurons. Ten μM antisense oligonucleotide of BMAL1 gene phase-dependently phase shifted the firing rhythm of individual SCN neurons. The result suggests the critical role of BMAL1 gene in the circadiant system at the level of the SCN neuron.2.Phenotype of mutation at the level of single SCN neuronsA slice including the SCN was dissected from newborn Clock mutant mice and spontaneous firing was monitored continuously for over 5 days. Irrespective of genotype, most SCN neurons in an SCN slice showed significant circadian firing rhythms. Mean circadian periods of firing rhythms in homozygous, heterozygous, and wild type mice were 27.2, 24.8 and 23.5 h, respectively. The periods were similar to the circadian period of wheel-running activity of each genotype mice. The amplitude of firing rhythms were not different among them. Present result clearly showed that Clock mutaion lengthend the period of individual SCN neurons, but did not affects the rhythm generation or its amplitude.

通过在多电极培养皿上培养新生大鼠或小鼠的视交叉上核（SCN）来检查单个视交叉上神经元的时钟基因功能。对单个神经元的自发放电进行长达数周的连续监测，并分析放电率的昼夜节律。1.BMAL1基因在单个SCN神经元昼夜放电节律中的功能将反义寡核苷酸应用于显示出稳健昼夜放电节律的SCN神经元，以及对节律参数（例如相位、周期和频率）的影响。幅度，在单个 SCN 神经元水平上进行了检查。 BMAL1 基因的 10 μM 反义寡核苷酸相位依赖性地改变单个 SCN 神经元的放电节律。结果表明BMAL1基因在SCN神经元水平的昼夜节律系统中发挥着关键作用。2.单个SCN神经元水平的突变表型从新生Clock突变小鼠身上切下包括SCN的切片，并连续监测5天以上的自发放电。无论基因型如何，SCN 切片中的大多数 SCN 神经元都显示出显着的昼夜节律。纯合子、杂合子和野生型小鼠的放电节律的平均昼夜周期分别为27.2、24.8和23.5小时。该周期与各基因型小鼠滚轮活动的昼夜节律周期相似。它们之间的放电节律的幅度没有差异。目前的结果清楚地表明，时钟突变延长了单个SCN神经元的周期，但不影响节律的产生或其幅度。