The higher-order brain dysfunction in TRH knockout mice

TRH 基因敲除小鼠的高阶脑功能障碍

• 批准号: 11671080
• 负责人: YAMADA Masanobu
• 金额: $ 2.24万
• 依托单位: Gunma University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671080/
• 关键词: TRH; knockout mouse; neurophysiological action; ノックアウトマウス

TRH has been reported to possess several neurophysiological actions in the brain. To gain insights into the molecular mechanisms underlying these effects, particularly in the cerebellum, we attempted to clone a cDNA that was regulated by TRH using TRH knockout mice and subtractive cDNA analysis. Over 100 clones obtained by subtractive hybridization analysis between the wild-type and TRH^<-/-> cerebellum were analyzed. Four clones among them were identical and cdc2-related kinase (PFTAIRE protein kinase 1 (PFTK1)) cDNA, which was previously reported to be expressed only in the brain and testis. PFTK1 mRNA levels in the euthyroid TRH^<-/-> cerebellum supplemented with thyroid hormone were significantly decreased compared with those in the wild-type. Induction of PFTK1 mRNA by TRH was also observed in a time- and dose-dependent manner in human medulloblastoma-derived HTB-185 cells that expressed TRH receptor subtype 1 mRNA. In addition, treatment of 8-Br-cGMP significantly increased PFTK1 mRNA levels, and a specific inhibitor of cGMP production, ODQ, completely blocked TRH-induced expression of PFTK1 mRNA. Furthermore, induction of PFTK1 mRNA by TRH was significantly inhibited by a NOS specific inhibitor, L-NAME, but not by a MEK inhibitor, PD98059 or a calcium channel inhibitor, nimodipine. These findings demonstrated, for the first time, a novel pathway between a neuropeptide and a cell cycle related peptide in the brain, and PFTK1 may be a key regulator for TRH action in the cerebellum through the NO-cGMP pathway.

据报道，TRH在大脑中具有多种神经生理学作用。为了深入了解这些作用的分子机制，特别是在小脑中，我们试图克隆一个cDNA，TRH基因敲除小鼠和消减cDNA分析TRH调节。分析了通过野生型和TRH^<-/->小脑之间的消减杂交分析获得的100多个克隆。其中4个克隆是相同的cdc 2相关激酶（PFTAIRE蛋白激酶1（PFTK 1））的cDNA，这是以前报道的只在大脑和睾丸中表达。补充甲状腺激素的甲状腺功能正常的TRH^<-/->小脑中PFTK 1 mRNA水平与野生型相比显著降低。在表达TRH受体亚型1 mRNA的人髓母细胞瘤衍生的HTB-185细胞中，也观察到TRH对PFTK 1 mRNA的诱导作用呈时间和剂量依赖性。此外，8-Br-cGMP处理显著增加PFTK 1 mRNA水平，cGMP产生的特异性抑制剂ODQ完全阻断TRH诱导的PFTK 1 mRNA表达。TRH对PFTK 1 mRNA的诱导作用可被NOS特异性抑制剂L-NAME显著抑制，但不被MEK抑制剂PD 98059或钙通道抑制剂尼莫地平抑制。这些发现首次证明了大脑中神经肽和细胞周期相关肽之间的新途径，PFTK 1可能是通过NO-cGMP途径在小脑中TRH作用的关键调节剂。

项目成果

Hashida T: "A novel TRH-PFTAIRE protein kinase 1 pathway in the cerebellum : subtractive hybridization analysis of TRH-deficient mice"Endocrinology. 143(7). 2808-2811 (2002)

Hashida T：“小脑中一种新的 TRH-PFTAIRE 蛋白激酶 1 通路：TRH 缺陷小鼠的消减杂交分析”内分泌学。

Yamada M: "Abundance of cyclo (His-Pro)-like immunoreactivity in the brain of TRH-deficient mice"Endocrinology. 140(1). 538-541 (1999)

Yamada M：“TRH 缺陷小鼠大脑中环 (His-Pro) 样免疫反应的丰度”内分泌学。

Shibusawa N.: "Requirement of thyrotropin-releasing hormone for the postnatal functions of pituitary thyrotrophs: ontogeny study of congenital tertiary hypothyroidism in mice"Mol Endocrinol.. 14(1). 137-146 (2000)

Shibusawa N.：“促甲状腺激素释放激素对垂体促甲状腺细胞出生后功能的需求：先天性三级甲状腺功能减退症小鼠的个体发育研究”Mol Endocrinol.. 14(1)。

Hashida T.: "A novel TRH-PFTAIRE protein kinase 1 pathway in the cerebellum: subtractive hybridization analysis of TRH-deficient mice"Endocrinology. 143(7). 2808-2811 (2002)

Hashida T.：“小脑中一种新型 TRH-PFTAIRE 蛋白激酶 1 通路：TRH 缺陷小鼠的消减杂交分析”内分泌学。

Shibusawa N: "Requirement of thyrotropin-releasing hormone for the postnatal functions of pituitary thyrotrophs : ontogeny study of congenital tertiary hypothyroidism in mice"Mol Endocrinol. 14(1). 137-146 (2000)

Shibusawa N：“垂体促甲状腺素出生后功能对促甲状腺素释放激素的需求：先天性三级甲状腺功能减退症小鼠的个体发育研究”Mol Endocrinol。