Abstract
This chapter describes the methods for isolation of nuclei from spermatozoa. The examples of methods for isolating sperm nuclei appropriate for organisms with various types of sperm histones are described. Some modifications suitable for other species are also discussed. Sperm histones of eutherian mammals are rich in arginine and stabilized by disulfide bonds formed from their cysteine residues. Protamines contain arginine residues in amounts up to two-thirds of the total amino-acid content and have been found in some metatherian mammals, birds, reptiles, (amphibians), fish, and molluscs. As the properties of sperm chromatin depend on their histones, methods for isolation of sperm nuclei differ from case to case. Structural and chemical differences between spermatozoa of different species and at different stages make necessary many modifications in methods for isolation of nuclei. In principle, spermatozoa are isolated from seminal ejaculate or reproductive tissue; sperm heads are separated from sperm tails, after which nonnuclear materials of the heads are removed. Nuclei of species that share similarities in properties of sperm chromatin with those for which satisfactory methods are available might also yield if subjected to similar procedures.
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