Vigodner M, Lewin L M, Shochat L, Mittelman L, Golan R
Department of Clinical Biochemistry, Sackler Medical School, Tel Aviv University, Ramat Aviv, Israel.
Mol Reprod Dev. 2003 Jan;64(1):86-95. doi: 10.1002/mrd.10212.
In this study, confocal microscopy and flow-cytometry were utilized to follow meiosis in hamster spermatogenesis. Confocal microscopy was used as an analytical tool to observe spermatocytes inside the tubules following meiotic progression consecutively at defined spermatogenic stages. To study spermatocyte differentiation, the structure of the synaptonemal complex was studied in detail at various stages of hamster spermatogenesis using the antibody against SC3 (the protein of axial/lateral element). The synaptonemal complex was observed from the leptotene stage until the first meiotic division with maximal staining in mid-pachytene spermatocytes, suggesting a role for SC3 at this postrecombinational stage. In addition, 3-dimensional (3D) images of synaptonemal complex were observed, providing information about spatial distribution of the chromosomes within the nuclei of spermatocytes at different stages of meiosis. Changes in spermatocyte sizes and DNA condensation allowed assessment of meiosis by flow cytometry. Changes in chromatin condensation at different stages of hamster meiosis were followed, revealing decondensation from early to late pachytene stages. The analysis also allowed a comparing of chromatin status of mitotic and meiotic chromosomes, confirming the less compact structure of the latter, possibly connected to increased transcriptional activity during meiosis.
在本研究中，共聚焦显微镜和流式细胞术被用于追踪仓鼠精子发生过程中的减数分裂。共聚焦显微镜被用作一种分析工具，以在特定的生精阶段连续观察减数分裂进程后曲细精管内的精母细胞。为了研究精母细胞的分化，使用抗SC3（轴/侧元件蛋白）抗体在仓鼠精子发生的各个阶段详细研究了联会复合体的结构。从细线期到第一次减数分裂都观察到了联会复合体，在粗线期中期的精母细胞中染色最强，这表明SC3在这个重组后阶段发挥作用。此外，还观察到了联会复合体的三维（3D）图像，提供了减数分裂不同阶段精母细胞核内染色体空间分布的信息。精母细胞大小和DNA凝聚的变化使得通过流式细胞术评估减数分裂成为可能。追踪了仓鼠减数分裂不同阶段染色质凝聚的变化，揭示了从粗线期早期到晚期的解凝聚过程。该分析还使得对有丝分裂和减数分裂染色体的染色质状态进行比较成为可能，证实了后者结构较松散，这可能与减数分裂期间转录活性增加有关。
