Molecular mechanisms of development of circadian rhythm in relation to transcription factor of peptide synthesis.

与肽合成转录因子相关的昼夜节律发育的分子机制。

• 批准号: 06807008
• 负责人: ISOBE Yoshiaki
• 金额: $ 1.34万
• 依托单位: Nagoya City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06807008/
• 关键词: Suprachiasmatic nuclei; Arg-vasopressin; Vasoactive intestinal peptide; GABA; Glutamate; Arg-Vasopressin; slice culture; 概日リズム; VAsoactive gntestinal peptide; gdstrin; glutamete; 自発行動リズム; 体温リズム

Mammalian circadian rhythum is regulated by a pacemaker in the suprachiasmatic nuclei (SCN) of hypothalamus. The SCN containing variety of peptides. Arg-vasopressin (AVP) containing neuron is one of the output neuron of circadian signal from the SCN to the other brain area. For the entraining mechanism of pacemaker, vasoactive intestinal peptide (VIP), gastrin-releasing peptide are considered to be concerned. While the function of these peptides on the AVP release is reciprocal ; i.s.we found that the VIP increased AVP release, while gastrin-releasing peptide decreased AVP release. Photic information is the most important factor for the entrainment, which is conveyed to SCN by glutamate containing neuron. Although the glutamate is higher in SCN during the dark period, glutamate plays dark pulse type phase response curve. Light pulse induces increment of glutamate at the ventrolateral area of SCN.So, we are examining now the mechanism of glutamate release from the SCN using a biosensor, which is continuously measurable and specific to glutamate.Beside, almost all of the SCN neuron containing GABA for an inhibitory amino-acid. GABA release shows diurnal rhythm that is higher at dark period than in light period. However the function of GABA is not clear yet. Therefore, we examined the effect of GABA on the AVP release using a slice culture system. The direct effect of GABA is not obvious, but the muscimol (GABAa-receptor agonist) increased the AVP level and bicuculline (GABAa antagonist) decreased AVP release, while the GABAb receptor agonist and/or antagonist did not show a significant effect.In preceding to analyze the pacemaking mechanism from the transcription factor of peptides, we are screening a SCN-specific, mRNA using differential display PCR.Many mRNA from SCN found to be expressed at a constant rate throughout a day. However, a few products appeared only at CT 20-23. We are analyzing the relationship with AP-1 and cAMP levels with them.

哺乳动物的昼夜节律是由下丘脑视交叉上核（SCN）中的起搏点调节的。SCN含有多种多肽。精氨酸加压素（AVP）神经元是SCN向其他脑区输出昼夜节律信号的神经元之一。对于起搏器的转运机制，血管活性肠肽（VIP）、胃泌素释放肽（GRP）被认为是值得关注的。而这些肽对AVP释放的作用是相互的，即我们发现VIP增加AVP释放，而胃泌素释放肽减少AVP释放。光信息是导致这种夹带的最重要因素，它通过含谷氨酸的神经元传递给SCN。暗期SCN中谷氨酸含量虽较高，但呈暗脉冲型相反应曲线。光脉冲可引起SCN腹外侧区谷氨酸的增加，因此，我们正在利用生物传感器研究SCN释放谷氨酸的机制，这种生物传感器对谷氨酸是连续可测量的和特异的。GABA的释放呈昼夜节律，暗期高于明期。但GABA的功能尚不清楚。因此，我们使用切片培养系统来检测GABA对AVP释放的影响。GABA的直接作用不明显，蝇蕈醇（GABA α受体激动剂）增加AVP水平和荷包牡丹碱（GABAa拮抗剂）减少AVP释放，而GABAb受体激动剂和/或拮抗剂没有显示出显著的效果。在从转录因子肽分析起搏机制之前，我们正在筛选SCN特异性，发现许多来自SCN的mRNA在一天中以恒定的速率表达。然而，少数产品仅在CT 20-23出现。分析AP-1和cAMP水平与它们的关系。

项目成果

Isobe,Y: "Ultradian and circadion rhythms of runnig wheel activity under the short light-dark cycle(0.5hr:0.5hr)in relation to light intensity and duration" Photomadicin and Photobiology. 16. 4-11 (1994)

Isobe，Y：“短光暗周期（0.5 小时：0.5 小时）下运行轮活动的超昼夜节律和昼夜节律与光强度和持续时间的关系”《Photomadicin 和光生物学》。

Isobe, Y., Kumazaki, M., Sakurai, T.and Nishino, H.: "Circadian rhythm of Arg-vasopressin in explant culture of rat suprachiasmatic nucleus : Effects of antibodies of vasoactive intestinal peptide and gastrin-releasing peptide." 4th IBRO congress, Kyoto,

Isobe, Y.、Kumazaki, M.、Sakurai, T. 和 Nishino, H.：“大鼠视交叉上核外植体培养物中精氨酸加压素的昼夜节律：血管活性肠肽和胃泌素释放肽抗体的影响”。

Isobe, Y.: "Ultradian and circadian rhythms of running wheel activity under the short light-dark cycle (0.5hm, 0.5hr) in relation to light intensity and duration" Photomed. Photobiol.16. 15-21 (1994)

Isobe, Y.：“短明暗周期（0.5 小时，0.5 小时）下跑轮活动的超昼夜节律和昼夜节律与光强度和持续时间的关系”Photomed。

Isobe,Y.: "Body temperatuie and Metabolism" T.Nagasaka amd A.S Milton(IPEC lnc,Tokyo), 234 (1995)

Isobe,Y.：“体温和新陈代谢”T.Nagasaka amd A.S Milton（IPEC lnc，东京），234（1995）

Isobe,Y: "Day-night differences in the contents of vasoactive intestinal peptide,gastrin-releaing peptide and Arg-vasopressin in the suprachiasmatic nucleus of rat pups cluring development" Neuros cience Letter. 187. 1-4 (1995)

Isobe，Y：“发育期大鼠视交叉上核中血管活性肠肽、胃泌素释放肽和精氨酸加压素含量的昼夜差异”《神经科学快报》。