Study of DNA integrity in somatic cells and spermatozoa by single-cell gel electrophoresis with silver-nitrate staining

Abstract

In the present work, genome instability in human and bovine peripheral blood lymphocytes and spermatozoa was studied by the method of DNA microelectrophoresis with subsequent staining of single cells with silver nitrate. A comparative analysis of the types of damage to human and bovine lymphocytes and spermatozoa genomes was performed. In the group of healthy donors, the spontaneous frequency of DNA damage revealed by single cell DNA microelectrophoresis did not exceed 9% and amounted, on average, to 4.8 ± 1.2%. In studying the effect of the duration of cryoconservation on bovine spermatozoa, no significant changes were revealed between the group of bulls whose spermatozoa were stored for less than one year (3.1 ± 0.9%) and the group of animals whose spermatozoa were under conditions of cryoconservation for more than 20 years (4.3 ± 0.5%). From the obtained single-cell DNA microelectropheretic data on the types and frequencies of DNA damage, a conclusion was made regarding the possibility of using a light variant based on cell staining with silver nitrate for the detection of genome instability, not only in somatic, but also in reproductive, cells.