He Xiao-jin, Yang Qing-ling, Pan Zhen-zhen, Wu Huan, Wang Liu, Shi Qing-hua, Zhao Ji-hua, Cao Yun-xia
Reproductive Medicine Center, the First Affiliated Hospital, Anhui Medical University, Hefei, Anhui 230022, P. R. China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2012 Jun;29(3):275-9. doi: 10.3760/cma.j.issn.1003-9406.2012.03.006.
To analyze defective homologous chromosomal recombination in Han Chinese azoospermic patients.
Testicular biopsy samples from 7 healthy controls and 7 Han Chinese azoospermic patients including 2 obstructive azoospermia (OA group) and 5 non-obstructive azoospermia (NOA group) were analyzed. Immunofluorescence staining was performed to categorize early stage cells at meiosis prophase and to analyze chromosome pairing and recombination of pachytene spermatocyte. Newly developed meiotic proteins antibodies (anti-SCP3, anti-synaptonemal complex proteins 3, anti-MLH1, anti-Mut-L Homolog 1, anti-CREST, chromosome centromere antibody) were used to identify synaptonemal complex (anti-SCP3), recombination sites (anti-MLH1) and centromere (anti-CREST), respectively. Staging of spermatocyte was determined according to SCP3 formation progression. Qualitative data were compared by a Chi-square test, and ANOVA was used to analyze quantitative data.
Respectively, 2346 and 2932 spermatocytes were categorized in the controls and azoospermic patients. The proportions of zygotene cells in both OA group and NOA group were significantly higher than that of the control group. Investigation of 1967 pachytene cells from the controls and 354 pachytene cells from azoospermic patients indicated that the mean MLH1 foci per pachytene cell of NOA group was statistically lower than that of the controls. Compared with the controls, incomplete synaptonemal complexes cells (containing gap and/or split) were significantly increased in the NOA group.
Delayed meiosis prophase is relatively common in azoospermic patients, and changes in quantity and distribution of recombination foci may be the cause for spermatogenesis arrest in Han Chinese population.
分析中国汉族无精子症患者中同源染色体重组缺陷情况。
对7名健康对照者以及7名中国汉族无精子症患者（包括2例梗阻性无精子症患者（OA组）和5例非梗阻性无精子症患者（NOA组））的睾丸活检样本进行分析。采用免疫荧光染色对减数分裂前期的早期细胞进行分类，并分析粗线期精母细胞的染色体配对和重组情况。使用新开发的减数分裂蛋白抗体（抗SCP3、抗联会复合体蛋白3、抗MLH1、抗Mut-L同源物1、抗CREST、染色体着丝粒抗体）分别鉴定联会复合体（抗SCP3）、重组位点（抗MLH1）和着丝粒（抗CREST）。根据SCP3形成进程确定精母细胞的分期。定性数据采用卡方检验进行比较，定量数据采用方差分析进行分析。
对照组和无精子症患者分别分类出2346个和2932个精母细胞。OA组和NOA组的偶线期细胞比例均显著高于对照组。对对照组的1967个粗线期细胞和无精子症患者的354个粗线期细胞进行研究表明，NOA组每个粗线期细胞的平均MLH1灶在统计学上低于对照组。与对照组相比，NOA组中不完全联会复合体细胞（包含间隙和/或分裂）显著增加。
减数分裂前期延迟在中国汉族无精子症患者中相对常见，重组灶数量和分布的变化可能是导致中国汉族人群精子发生停滞的原因。
