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

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 A(3) (CMA(3)) 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 CMA(3) staining, sperm from one bull was subjected to deprotamination of nuclei. The percentage of CMA(3)-positive bovine sperm did not vary among batches. Only two bulls showed a higher percentage of CMA(3)-positive sperm cells compared to the others. CMA(3) is a simple and useful tool for detecting sperm protamine deficiency in bulls.