Modification of brain function regulating metabolism by antisense oligodeoxynucleotides

反义寡脱氧核苷酸调节代谢的脑功能的改变

• 批准号: 07506004
• 负责人: TAKAHASHI Michio
• 金额: $ 8.58万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07506004/
• 关键词: Antisense oligodeoxynucleotides; Hypothalamus; Metabolism; Feeding behavior; Obesity; Running activity; gamma-aminobutyric acid; glutamic acid decarboxylase; γ-アミノ酪酸

In the ventromedial nucleus of the hypothalamus (VMH), gamma-aminobutyric acid (GABA) plays an important role in regulating metabolism. The GABA synthetic enzyme, glutamic acid decarboxylase (GAD) consists of two isozymes, GAD65 and GAD67. In the present study, the effect of injection of antisense oligodeoxynucleotides (ODNs) to each GAD isozyme into the VMH of the male rat on food intake, body weight and locomotor activity was examined. Phosphorothioated antisense ODNs were incorporated in the water-absorbent polymer (WAP) and injected into bilateral VMH,and the day of injection was designated as day 0. Antisense ODNs to GAD65 and GAD67 tended to reduce food intake on day 1, though not significant. Simultaneous injection of antisense ODNs to both GAD isozymes significantly decreased food intake on only day 1, but body weight remained suppressed by this treatment for 5 days. On the other hand, simultaneous administration of antisense ODNs to GAD65 and GAD67 significantly increased loco … More motor activity between days 3-5, whcreas other treatments did not. This increase in daily locomotor activity was mainly due to the increase in locomotor activity during the light, but not dark, phase. Neither vehicle (WAP) nor control ODN affected food intake, body weight and locomotor activity. Histological studies indicated that, 24 h after the injection, antisense ODN distributed within about 800 mum from the edge of area where WAP was located and then gradually disappeared with days, but still remained within 300 mum distance even 7 days after the injection. Antisense ODN was effectively incorporated by all the cell types examined, i.e.neurons, astrocytes and microglias. Further, HPLC analysis revealed that antisense ODN to either GAD isozyme decreased the content of GABA,but not other amino acids, in the VMH by 24 h. These results suggest that either GAD isozyme is synergistically involved in enhancing food intake and suppressing locomotor activity in rat VMH.This study also indicates that central injection of antisense oligodeoxynucleotides is useful means to regulate animal metabolism. Less

在下丘脑腹内侧核（VMH）中，γ-氨基丁酸（GABA）在调节代谢中发挥着重要作用。 GABA 合成酶谷氨酸脱羧酶 (GAD) 由两种同工酶 GAD65 和 GAD67 组成。在本研究中，检查了将每种GAD同工酶的反义寡脱氧核苷酸（ODN）注射到雄性大鼠的VMH中对食物摄入、体重和运动活性的影响。将硫代磷酸化反义 ODN 掺入吸水性聚合物 (WAP) 中并注射到双侧 VMH，注射日指定为第 0 天。针对 GAD65 和 GAD67 的反义 ODN 在第 1 天倾向于减少食物摄入量，但并不显着。同时注射两种 GAD 同工酶的反义 ODN 仅在第一天就显着降低了食物摄入量，但这种治疗方法在 5 天的时间里体重仍然受到抑制。另一方面，同时给予 GAD65 和 GAD67 反义 ODN 显着增加了第 3-5 天之间的局部运动活动，而其他治疗则没有。每日运动活动的增加主要是由于亮阶段运动活动的增加，而不是暗阶段的运动活动的增加。媒介物 (WAP) 和对照 ODN 均不影响食物摄入、体重和运动活动。组织学研究表明，注射后24小时，反义ODN分布在距WAP所在区域边缘约800μm的范围内，随后逐渐消失，但注射后7天仍保持在300μm的距离内。反义 ODN 被所有检查的细胞类型（即神经元、星形胶质细胞和小胶质细胞）有效地掺入。此外，HPLC 分析显示，GAD 同工酶的反义 ODN 降低了 VMH 中 GABA 的含量，但没有降低其他氨基酸的含量 24 小时。这些结果表明，任一GAD同工酶协同参与增强大鼠VMH的食物摄入和抑制运动活性。该研究还表明，中枢注射反义寡脱氧核苷酸是调节动物代谢的有用手段。较少的

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