使用放线菌素A3染色检测牛精子细胞中的鱼精蛋白缺乏情况。

Use of chromomycin A3 staining in bovine sperm cells for detection of protamine deficiency.

作者信息

Simões R, Feitosa W B, Mendes C M, Marques M G, Nicacio A C, de Barros F R O, Visintin J A, Assumpção M E O A

机构信息

Laboratory of in Vitro Fertilization, Cloning and Animal Transgenesis, Department of Animal Reproduction, School of Veterinary Medicine and Animal Sciences, University of Sao Paulo, Sao Paulo, Brazil.

出版信息

Biotech Histochem. 2009 Jun;84(3):79-83. doi: 10.1080/10520290902843595.

DOI:10.1080/10520290902843595

PMID:19306222

Abstract

Sperm chromatin integrity is essential for accurate transmission of male genetic information, and normal sperm chromatin structure is important for fertilization. Protamine is a nuclear protein that plays a key role in sperm DNA integrity, because it is responsible for sperm DNA stability and packing until the paternal genome is delivered into the oocyte during fertilization. Our aim was to investigate protamine deficiency in sperm cells of Bos indicus bulls (Nelore) using chromomycin A3 (CMA3) staining. Frozen semen from 14 bulls were thawed, then fixed in Carnoy's solution. Smears were prepared and analyzed by microscopy. As a positive control of CMA3 staining, sperm from one bull was subjected to deprotamination of nuclei. The percentage of CMA3-positive bovine sperm did not vary among batches. Only two bulls showed a higher percentage of CMA3-positive sperm cells compared to the others. CMA3 is a simple and useful tool for detecting sperm protamine deficiency in bulls.

摘要

精子染色质完整性对于准确传递雄性遗传信息至关重要，而正常的精子染色质结构对受精也很重要。鱼精蛋白是一种核蛋白，在精子DNA完整性方面起着关键作用，因为它负责精子DNA的稳定性和包装，直到受精时父本基因组被传递到卵母细胞中。我们的目的是使用嗜铬霉素A3（CMA3）染色法研究印度瘤牛（Nellore）精子细胞中的鱼精蛋白缺乏情况。将来自14头公牛的冷冻精液解冻，然后固定在卡诺氏溶液中。制备涂片并通过显微镜进行分析。作为CMA3染色的阳性对照，对一头公牛的精子进行细胞核脱鱼精蛋白处理。不同批次的CMA3阳性牛精子百分比没有差异。只有两头公牛的CMA3阳性精子细胞百分比高于其他公牛。CMA3是检测公牛精子鱼精蛋白缺乏的一种简单而有用的工具。