Research for modulation of expression of serotonin transporter and its involvement in mental disorder

血清素转运蛋白表达调节及其与精神障碍的关系研究

• 批准号: 09670091
• 负责人: SAITO Naoaki
• 金额: $ 2.05万
• 依托单位: KOBE UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670091/
• 关键词: serotonin; transporter; interferon; phosphorylation; depression; synapse; electron microscopy; 脱リン酸化; 変異体; COS-7細胞

We examined the regulatory mechanism of serotonin transporter and the involvement of serotonin transporter (SET) in depression to elucidate the function role of SET in mental disorder.1. Regulation of SET by phosphorylation. The regulation of SET expressed in COS7 cells by the activation of protein kinase C with phorbol ester or by inhibition of protein phosphatases with calyculin A was examined. Both treatment decreased the activity of SET with a reduction in Vmax without affecting Km. Site-directed mutagenesis of five putative PKC phosphorylation sites in SET revealed that these inhibitory modulation of SET activity did not act via direct phosphorylation of SET by PKC.2. Immunocytochemical localization of SET.The cellular and intracellular localization of SET was examined by immunocytochemistry. SET was localized in the varicose nerve fibers and nerve terminals but not in the cell bodies. Under electron microscopy, SET was seen around the synaptic vesicles as well as presynaptic membrane.3. The effects of interferon-alpha and -gamma on the transcription of SET.Both SET mRNA and SET activity were increased by treatment with interferon-alpha and -gamma for 3 h. Treatment with dibutyryl cAMP also increased SET mRNA and SET activity. The level of SET mRNA was increased both in the midbrain and adrenal glands of mice which were treated with interferons for 3 h. These results suggest that the interferon-induced psychiatric side effects arise through regulation of serotonin transporter transcription and that the transcriptional regulation of the serotonin transporter is a possible neurochemical mechanism of affective disorders.

我们研究了5-羟色胺转运蛋白的调节机制以及5-羟色胺转运蛋白（SET）在抑郁症中的参与，以阐明SET在精神障碍中的功能作用。1．通过磷酸化调节 SET。研究了通过佛波酯激活蛋白激酶 C 或通过花萼蛋白 A 抑制蛋白磷酸酶来调节 COS7 细胞中表达的 SET。两种治疗均降低了 SET 活性，同时降低了 Vmax，但不影响 Km。 SET 中五个假定的 PKC 磷酸化位点的定点诱变表明，这些 SET 活性的抑制性调节并不是通过 PKC.2 对 SET 的直接磷酸化来发挥作用。 SET的免疫细胞化学定位。通过免疫细胞化学检查SET的细胞和细胞内定位。 SET 定位于曲张的神经纤维和神经末梢，但不定位于细胞体。电镜下突触小泡周围及突触前膜可见SET。 3．干扰素-α和干扰素-γ对SET转录的影响。干扰素-α和-γ处理3小时后SET mRNA和SET活性均增加。二丁酰 cAMP 处理也增加了 SET mRNA 和 SET 活性。用干扰素处理3小时的小鼠的中脑和肾上腺中SET mRNA的水平均增加。这些结果表明，干扰素诱导的精神副作用是通过调节血清素转运蛋白转录而产生的，并且血清素转运蛋白的转录调节是情感障碍的可能的神经化学机制。

项目成果

Hashimoto,T.他: "Isoform-specific redistribution of calcineurin Aα and Aβ in hippocampal CA1 region of Gerbil after transient ischemia." J.Neurochem.70. 1289-1298 (1998)

Hashimoto, T. 等人：“瞬时缺血后沙鼠海马 CA1 区钙调神经磷酸酶 Aα 和 Aβ 的异构体特异性重新分布。J.Neurochem.70 (1998)”

Obata H.et al.: "GABAergic neurotransmission in rat taste buds ; Immunocytochemical study for GABA and GABA transporter subtypes." Mol.Brain Res.49. 29-36 (1997)

Obata H.等人：“大鼠味蕾中的 GABA 能神经传递；GABA 和 GABA 转运蛋白亚型的免疫细胞化学研究。”

Ohmori, S.et al.: "Three distinct mechanisms for translocation and activation of delta-subspecies of protein kinase C." Mol.Cell.Biol.18. 5263-5271 (1998)

Ohmori, S.et al.：“蛋白激酶 C δ 亚种易位和激活的三种不同机制。”

Nakashita M., 他: "Effects of tricyclic and tetracyclic antidepressants on the three subtypes of GABA transporter." Neurosci.Res.29. 87-91 (1997)

Nakashita M. 等人：“三环类和四环类抗抑郁药对 GABA 转运蛋白三种亚型的影响”，Neurosci.Res.29 (1997)。

Hashimoto T.et al.: "Isoform-specific redistribution of calcineurin Aalpha and Abeta in hippocampal CA1 region of Gerbil after transient ischemia." J.Neurochem.70. 1289-1298 (1998)

Hashimoto T.等人：“短暂性缺血后，沙鼠海马 CA1 区钙调神经磷酸酶 Aalpha 和 Abeta 的异构体特异性重新分布。”