Alam Md Hasanur, Qiao Huanyu
Department of Comparative Biosciences, University of Illinois Urbana-Champaign, Urbana, IL, USA.
Methods Mol Biol. 2025;2954:199-206. doi: 10.1007/978-1-0716-4698-4_10.
Meiosis, a special cell division, occurs in germ cells to produce gametes. It is vital for reproduction and fostering genetic diversity in offspring. Understanding and characterizing meiotic events are essential for elucidating genetic disorders. Here, we demonstrate a classical technique employing mouse spermatocyte chromosome spreads to study mammalian meiosis. Spermatocytes are isolated from seminiferous tubules of the testis and gently released on a depression slide. These cells are then immersed in a hypotonic solution to introduce swelling. Subsequent incubation in a sucrose solution facilitates optimally spreading nuclei over a glass slide, aided by the application of a fixative. To examine the key meiotic events, such as synapsis and homologous recombination, the chromosome spreads undergo immunolabeling with both primary and secondary antibodies. This method offers a robust framework for studying the intricacies of meiosis in mammals, providing valuable insights into the mechanisms of reproductive biology.
减数分裂是一种特殊的细胞分裂，发生在生殖细胞中以产生配子。它对于繁殖和促进后代的遗传多样性至关重要。理解和表征减数分裂事件对于阐明遗传疾病至关重要。在这里，我们展示了一种利用小鼠精母细胞染色体铺展来研究哺乳动物减数分裂的经典技术。从睾丸的生精小管中分离出精母细胞，并将其轻轻放置在凹玻片上。然后将这些细胞浸入低渗溶液中使其膨胀。随后在蔗糖溶液中孵育有助于在玻片上最佳地铺展细胞核，这借助于固定剂的应用。为了检查关键的减数分裂事件，如联会和同源重组，染色体铺展需用一抗和二抗进行免疫标记。该方法为研究哺乳动物减数分裂的复杂性提供了一个强大的框架，为生殖生物学机制提供了有价值的见解。
