标记减数分裂遗传重组的免疫荧光染色方法

联会复合体免疫荧光技术在全基因减数分裂遗传重组研究中具有精确和直观的优势.本研究通过免疫荧光染色方法制备小鼠精母细胞联会复合体,研究其形态组成与遗传重组特征,展示雄性小鼠遗传重组图谱并分析其重组位点(MLH1位点)的分布特征.4只小鼠共145个精母细胞在平均每个细胞的MLH1位点数为23.3±2.4;在常染色体联会复合体中,未发现有3个MLH1位点的联会复合体,具有1个MLH1位点的联会复合体较多,平均为14.2;无XY联会复合体的细胞占所有细胞的4.1%,XY联会复合体上有MLH1位点的细胞占30.2%;联会复合体上有裂缝的细胞占0.7%.通过联会复合体免疫荧光染色可以清晰地分辨出联会复合体(红色)、着丝粒(蓝色)和MLH1位点(绿色),是遗传重组分析的一种强有力工具.

Immunofluorescent Staining Used for Marking Meiotic Recombination

Immunofluorescence techniques make the analysis of meiotic recombination across the whole genome more precise and the visualization of some important meiotic structures on synaptonemal complexes (SC) more distinguishable. Immunofluorescent staining was employed to prepare SCs of mouse spermatocytes. Meiotic recombination patterns of male mouse were exhibited by examing the distribution features of DNA mismatch repair protein (MLH1 foci). The mean number of MLH1 foci per cell in 145 cells from four individuals was 23.3±2.4 and none of the autosomal SCs was found to be with three MLH1 foci. However, the SCs with one MLH1 foci per cell dominated in autosomal SCs with an average of 14.2. The cells with asynapsed XY and MLH1 foci averaged 30.2% in the cells analyzed, while 4.1% of the cells exhibited no asynapsed XY. The percentage of cells with gaps was 0.7%. The SCs, centromere and MLH1 foci were immunofluorescently stained red, blue and green, respectively, as an effective technique to investigate meiotic recombination.