Study of nuclear membrane differentiation between germ and vegetative nuclei in early development of Paramecium.

草履虫早期发育过程中胚芽与营养细胞核膜分化的研究。

• 批准号: 02804063
• 负责人: FUJISHIMA Masahiro
• 金额: $ 1.09万
• 依托单位: Yamaguchi University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02804063/
• 关键词: Paramecium; Macronucleus; Nuclear membrane; Nuclear membrane antigen; Nuclear membrane differentiation; Monoclonal antibody; Nuclear differentiation; 栄養核; 核膜特異的抗原; 繊毛; 接合型物質

1. Isolated macronuclei of Paramecium caudatum were injected into mice, and hybridomas bet ween the spleen cells and mieroma cells (NS-1) were obtained by cell fusion. The hybridoma was screened by indirect immunofluorescence microscopy and cloned by limiting dilution. Monoclonal antibody (MAM-1) against macronuclear membrane of P. caudatum was obtained from cell-fee medium of the hybridoma cells.2. Molecular weight of the MAM-1 antigen was ca. 70 kDa by immunoblotting.3. Indirect immunofluorescence microscopy showed that the antigens appeared in 4 out of 8 postzygotic nuclei immediately after the 4 nuclei differentiated morphologically into the macronuclear anlagen, and the antigens were constitutionally present in the membrane through the cell cycle. The antigens were not detected in the micronucleus. The results indicate that the nuclear membranes differentiate between the two kinds of nuclei in early development.4. Immunofluorescence microscopy on species-specificity of the cross reactivity of the antibody showed that all strains of P. caudatum and P. jenningsi examined showed the flurescence in the macronuclear membranes but not in P. bursaria and Tetrahymena thermophila and T. pyriformis examined.

1.将分离的草履虫大核注入小鼠体内，用细胞融合法获得小鼠脾细胞与瘤细胞（NS-1）的杂交瘤。用间接免疫荧光显微镜筛选杂交瘤细胞，有限稀释法克隆。从杂交瘤细胞的细胞培养液中获得抗尾状核单克隆抗体（MAM-1）. MAM-1抗原的分子量约为1000。70 kDa的免疫印迹。间接免疫荧光显微镜观察显示，在8个合子后细胞核中，有4个在形态上分化为大核原基后立即出现抗原，并在整个细胞周期中组成性地存在于细胞膜上。微核中未检测到抗原。结果表明，两种细胞核的核膜在发育早期就有区别.用免疫荧光显微镜观察抗体的种特异性，结果表明，尾拟单胞菌和詹氏拟单胞菌的大核膜均显示荧光，而囊拟单胞菌、嗜热四膜虫和T.梨形肌检查。

项目成果

M.Fujishima: "Developmental expression of macronuclear specific antigen in Paramecium caudatum." Developmental Genetics. (1992)

M.Fujishima：“尾草履虫大核特异性抗原的发育表达。”

M. Fujishima: "Injection of MRIF of ciliate Tetrahymena thermophila into amphibian Bufo and Xenopus oocytes induces MPF activity by dephosphorylation of the oocyte cdc2." Journal of Protozoology.

M. Fujishima：“将纤毛虫嗜热四膜虫的 MRIF 注射到两栖动物蟾蜍和爪蟾卵母细胞中，通过卵母细胞 cdc2 的去磷酸化诱导 MPF 活性。”

M. Fujishima: "Induction of reinitiation of meiosis in amphibian Bufo and Xenopus oocytes by injection of M-phase extracts of ciliate Tetrahymena needs the recipient protein synthesis." Experimental Cell Research. 193. 155-160 (1991)

M. Fujishima：“通过注射纤毛虫四膜虫的 M 期提取物来诱导两栖动物蟾蜍和非洲爪蟾卵母细胞减数分裂的重新启动，需要受体蛋白质的合成。”

M. Fujishima: "Developmental expression of macronuclear specific antigen in Paramecium caudatum." Developmental Genetics.

M. Fujishima：“尾草履虫大核特异性抗原的发育表达。”

M. Fujishima: "Costimulation-induced rounding in Tetrahymena thermophila : Early cell-shape transformation induced by sexual cell-to-cell collisions between complementary meting types." Developmental Biology.

M. Fujishima：“共刺激诱导的嗜热四膜虫变圆：互补交配类型之间有性细胞间碰撞诱导的早期细胞形状转变。”