Autonomic nerve and integration of biological clock signal

使用大鼠，我们得到了以下结果。A)血糖控制：视交叉上核（SCN）中的血管活性肠肽(VIP) -神经元和精氨酸加压素(AVP) -神经元是昼夜节律的主振荡器，被认为通过增强交感神经参与血糖控制。B)神经通路：利用假狂犬病毒在神经元中逆行转运，我们获得了SCN通过交感和副交感传出神经多突触向胰腺传递神经信息的证据，这两条神经通路都经过下丘脑室旁核和纹带。C)发现2个新蛋白（SCOP和B10）在SCN中表达，SCOP（1696个氨基酸）包含pleckstrin同源结构域、富含亮氨酸的重复序列和蛋白磷酸酶2c样结构域，其在SCN中的表达具有昼夜节律性。此外，在SCN中注入SCOP mRNA的反义寡核苷酸可降低SCN中SCOP的表达，并消除运动活动的昼夜节律。而在COS-7细胞中，B10的过表达导致Per1-3与这些蛋白结合后进入细胞核。D)分子机制：Buijs和他的同事发现了Cry1和Cry2参与哺乳动物昼夜节律振荡机制的证据。因此，我们尝试制造针对这些和Per1-3的抗体。目前我们已经获得了针对Cry1， Per1和Per2的抗体，现在我们正试图研究这些蛋白在自主神经和生物钟整合机制中的作用。

Using rats we obtained following results. A) Control of blood glucose : vasoactive intestinal peptide (VIP) -neurons and arginine vasopressin (AVP) -neurons in the suprachiasmatic nucleus (SCN), a master circadian oscillator, are suggested to be involved in the control of blood glucose by enhancing the sympathetic nerves. B) Neural pathways : using pseudorabies virus which is retrogradely transported in neurons, we obtained evidence that the SCN multisynaptically send neural informations to the pancreas through both sympathetic and parasympathetic efferent nerves, and that both neural pathways pass the hypothalamic paraventricular nucleus and zona incerta. C) Discovery of two novel proteins : We found 2 novel proteins (SCOP and B10) expressed in the SCN, and that SCOP (1696 amino acids) contains pleckstrin homology domain, leucine-rich repeats and protein phosphatase 2C-like domain and showed a circadian rhythm in its expression in the SCN. Moreover, it was observed that infusions of antisense oligo to SCOP mRNA into the SCN reduced the expression of SCOP in the SCN and eliminated the circadian rhythm of locomotor activity. While, the overexpression of B10 in COS-7 cells caused nuclear entries of Per1-3 after bining with these proteins. D) Molecular machanism : Buijs and his colleags found evidence that Cry1 and Cry2 are involved in the mechanism of mammalian circadian oscillation. Thus, we tried to make antibodies against these and Per1-3. So far we obtained antibodies against Cry1, Per1 and Per2, and now we are trying to examine roles of these proteins in the integration mechanism of autonomic nerves and circadian clock.