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Adipocytes as endocrine cells and their roles in mice (PPARgamma knockout mice)

脂肪细胞作为内分泌细胞及其在小鼠中的作用（PPARgamma 敲除小鼠）

基本信息

批准号：
08557063
负责人：
KASUGA Masato
金额：
$ 10.37万
依托单位：
Kobe University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

项目摘要

To elucidate the physiological function of adipocytes in intact animals, we tried to make PPAR_<gamma> knockout mice. However, during our experiments, we were informed that PPAR_<gamma> knockout mice were embryonic lethal. therefore, we tried another approach. That is, adipose tissue-specific expression of dominant-negative mutants of PPAR_<gamma>, To find dominant-negative mutants of PPAR_<gamma>, we made several mutants and expressed in 3T3-L1 preadipocytes using adenovirus vector and found that the mutant, in which 16 amino acids in C-terminal were deleted, inhibited the adipocyte differentiation induced by thiazolidinedione. Therefore, this mutant may be useful to make fat-less mice by the transgenic method.Since PPAR_<gamma> has an important role in adipocyte differentiation and systemic insulin action, functional defects in PPAR_<gamma> might be expected to result in the impaired adipogenesis and insulin resistance, conditiones typical of lipoatrophic diabetes. Therefore, we examined the mutation in the coding sequences of PPAR_<gamma> gene by PCR-SSCP and found that the coding sequences of the PPAR_<gamma> gene are normal in individuals with lipoatrophic diabetes.Furthermore, we also examined the mutation in the coding sequences of the PPAR_<gamma> gene in individuals with type 2 diabetes and found the Prol2Ala mutations. However, the allele frequency of this mutation between normal and type2 diabetic patiants were not changed.

为了阐明正常动物脂肪细胞的生理功能，我们尝试制造PPAR基因<gamma>敲除小鼠。然而，在我们的实验中，我们被告知，PPAR_1基因<gamma>敲除小鼠是胚胎致死的。所以我们尝试了另一种方法。为了寻找PPAR_1的显性失活突变体，<gamma>我们构建了PPAR_1的显性失活突变体<gamma>，并利用腺病毒载体在3 T3-L1前脂肪细胞中表达，发现C端16个氨基酸缺失的突变体抑制噻唑烷二酮诱导的脂肪细胞分化。由于PPAR_1<gamma>在脂肪细胞分化和全身性胰岛素作用中具有重要作用，PPAR_1功能缺陷<gamma>可能导致脂肪形成障碍和胰岛素抵抗，这是脂肪萎缩性糖尿病的典型症状。因此，我们用PCR-SSCP方法检测了脂肪萎缩性糖尿病患者的PPAR_1基因编码序列的突变<gamma>，发现<gamma>脂肪萎缩性糖尿病患者PPAR_1基因编码序列正常，同时检测了<gamma>2型糖尿病患者PPAR_1基因编码序列的突变，发现Pro 12 Ala突变。2型糖尿病患者与正常对照组相比，其等位基因频率无明显变化。