Analysis of sinnalling pathway of apoptosis degradation using PKCδ gene-targeting mice

PKCδ基因靶向小鼠分析细胞凋亡降解信号通路

• 批准号: 09480189
• 负责人: NAKAYAMA Kei-ichi
• 金额: $ 8.83万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 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-09480189/
• 关键词: PKCδ; apoptosis; knockout mice; singal transduction; B cell; proliferation control; プロテインキナーゼC; ジーンターゲティング; ES細胞; ICE様プロテアーゼ

It has been reported that protein kinase Cδ (PKCδ) a subtype of PKC family members, is cleaved by caspase resulting in conversion to activated PKCδ upon apoptosis. The aim of this research project is to elucidate the physiological function of PKCδ by generating mice deficient in PKCδ gene.We constructed the targeting vector and transfect it into embryonic stem (ES) cells to obtain the homologous recombinant of ES cells for PKCδ locus. According to the standard method to generate gene-ablated mice, PKCδ-deficient mice were produced with the mutant ES cells. The PKCδ-deficient mice were viable, healthy, and not prone to cancer development.Histlogical examinations of the PKCδ-deficient mice revealed splenomegaly due to abnormal accumulation of T and B lymphocytes. In lymph nodes, increase in the number of B cells was more apparent than that of T cells. In vitro culture experiments showed that PKCδ-deficient B cells grew faster than wild-type B cells.We concluded that PKCδ is not essential for the apoptosis during development, because of the lack of developmental abnormalities. Furthermore, apoptosis of thymovytes and embryonic fibroblasts from PKCδ-deficient mice seems unaffected. However, the extent of apoptosis in B cells appeared to be reduced, suggesting that PKCδ may play an important role in the signaling pathway of apoptosis in B cells.

据报道，蛋白激酶 Cδ (PKCδ) 是 PKC 家族成员的一种亚型，被 caspase 裂解，导致细胞凋亡时转化为激活的 PKCδ。本研究项目的目的是通过产生缺乏 PKCδ 的小鼠来阐明 PKCδ 的生理功能。 PKCδ基因。按照标准方法构建靶向载体，转染胚胎干细胞，获得PKCδ基因座的ES细胞同源重组体。为了生成基因敲除小鼠，我们用突变型 ES 细胞培育出了 PKCδ 缺陷小鼠。PKCδ 缺陷小鼠能够存活、健康且不易患癌症。PKCδ 缺陷小鼠的组织学检查显示，由于异常积累，脾脏肿大。在淋巴结中，B 细胞数量的增加比 T 细胞更明显。 体外培养实验表明，PKCδ 缺陷的 B 细胞比野生型 B 细胞生长得更快。我们得出结论，由于缺乏发育异常，PKCδ 对于发育过程中的细胞凋亡不是必需的。此外，PKCδ 缺陷小鼠的胸腺细胞和胚胎成纤维细胞的凋亡似乎不受影响，但 B 细胞的凋亡程度似乎不受影响。降低，提示 PKCδ 可能在 B 细胞凋亡信号通路中发挥重要作用。

项目成果

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