Physiological analysis of the role of PACAP using transgenic and knockout mice

使用转基因和基因敲除小鼠对 PACAP 作用的生理分析

• 批准号: 12470016
• 负责人: BABA Akemichi
• 金额: $ 8.38万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470016/
• 关键词: PACAP; gene knockout mice; transgenic mice; psychomotor behavior; pain; circadian rhythm; diabetes; β-cell neogenesis; 遺伝子改変マウス; 精神疾患モデル動物; 海馬機能; 神経因性・炎症性疼痛; 好奇心; 不安; 神経細胞保護; グルタミン酸

This study examined the physiological and pathophysiological roles of endogenous PACAP in the central and peripheral nervous system using PACAP gene knockout (PACAP^<-/->) mice, transgenic mice overexpressing PACAP in the pancreas (PACAP-Tg), and PACAP (PAC_1) receptor exon 2-targeted (PAC_1^<-/->) mice which were recently generated in this laboratory. Through these in vivo mutagenesis studies, we obtained the following results.1. Psychomotor function(1) PACAP^<-/-> mice exhibited hyperactive and explosive jumping behaviors in an open field, and increased exploratory behavior and less anxiety. These aberrant behaviors are ameliorated by the antipsychotic drug and SSRI.(2) PACAP^<-/-> mice exhibited a significant impairment of prepulse inhibition of the acoustic startle response.These results provide evidence that PACAP plays a previously uncharacterized role in the regulation of psychomotor behaviors and suggest that the phenotype of PACAP^<-/-> mice in part resembles those in human sc … More hizophrenia and/or attention deficit/hyperactivity disorder (ADHD).2. Hippocampal functionPACAP^<-/-> mice and PAC_1^<-/-> mice showed an impairment of hippocampal long-term potentiation (LTP). In addition, the behavioral study revealed a deficit in hippocampus-dependent associative learning, implicating PACAP-ergic neurons in regulation of higher brain functions.3. Pain transmissionAllodynia and hyperalgesia in the neuropathic and inflammatory pain models were significantly reduced in PACAP^<-/-> mice.4. Circadian rhythmThe light-induced phase resetting and c-Fos expression in the suprachiasmatic nucleus (SCN) were attenuated in PACAP^<-/-> mice, highlighting the in vivo role of PACAP in circadian photoentrainment.5. Glucose and insulin homeostasis(1) The PACAP transgene ameliorated streptozotocin-induced diabetes in PACAP-Tg mice. Notably, an increase in 5-bromo-2-deoxyuridine (BrdU)-positive β cells in the streptozotocin-treated PACAP-Tg mice was observed.(2) We crossed PACAP-Tg and lethal yellow KKA^y mice, a genetic model for obesity-diabetes. KKA^y mice showed marked hyperglycemia associated with compensatory hyperinsulinaemia and islet hyperplasia; however, both hyperinsulinemia and islet hyperplasia were significantly suppressed in KKA^y mice carrying the PACAP transgene.The present data suggest a protective role for PACAP against type I diabetes as well as obesity-associated (type II) diabetes, and the possible involvement of PACAP in the β cell neogenesis. This is the first report suggesting in vivo roles of PACAP on islet hyperplasia and β cell neogenesis, providing the possibility that drugs associated with PACAP-signaling pathways might be of therapeutic value for the treatment of diabetes. Less

本研究采用PACAP基因敲除（PACAP^<-/->）小鼠、胰腺中过表达PACAP的转基因小鼠（PACAP-Tg）和本实验室最近培育的PACAP（PAC_1）受体外显子2靶向（PAC_1^<-/->）小鼠，研究内源性PACAP在中枢和外周神经系统中的生理和病理生理作用。通过这些体内诱变研究，我们获得了以下结果.心理功能（1）PACAP^<-/->小鼠在开阔场地表现出过度活跃和爆发性跳跃行为，探索行为增加，焦虑减少。抗精神病药物和SSRI可以改善这些异常行为。(2)PACAP^<-/->小鼠表现出对声惊吓反应的前脉冲抑制的明显损害。这些结果提供了证据，表明PACAP ^<-/->小鼠在精神行为的调节中起着以前未被表征的作用，并表明PACAP^<-/->小鼠的表型部分类似于人类的表型。 ...更多信息 多动症和/或注意力缺陷/多动障碍（ADHD）。PACAP ^<-/->小鼠和PAC ^<-/->小鼠海马长时程增强（LTP）功能受损。此外，行为学研究揭示了海马区依赖性联想学习的缺陷，暗示PACAP能神经元参与了高级脑功能的调节.疼痛传递PACAP^<-/->小鼠神经病理性疼痛和炎症性疼痛模型中的异常性疼痛和痛觉过敏显著降低.昼夜节律在PACAP^-/->小鼠中，光诱导的相位重置和视交叉上核（SCN）中的c-Fos表达减弱，突出了PACAP在昼夜光调节中的体内作用。（1）PACAP转基因改善了链脲佐菌素诱导的PACAP-Tg小鼠糖尿病。值得注意的是，在链脲佐菌素处理的PACAP-Tg小鼠中观察到5-溴-2-脱氧尿苷（BrdU）阳性β细胞增加。(2)我们将PACAP-Tg和致命的黄色KKA^y小鼠（肥胖糖尿病的遗传模型）杂交。KKA^y小鼠表现出明显的高血糖，并伴有代偿性高胰岛素血症和胰岛增生;然而，携带PACAP转基因的KKA^y小鼠的高胰岛素血症和胰岛增生均受到显着抑制。目前的数据表明PACAP对I型糖尿病以及肥胖相关（II型）糖尿病具有保护作用，并且PACAP可能参与β细胞新生。这是第一份表明PACAP对胰岛增生和β细胞新生的体内作用的报告，提供了与PACAP信号通路相关的药物可能对糖尿病治疗具有治疗价值的可能性。少

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