Diploid spermatozoa caused by failure of the second meiotic division in a bull

An artificial insemination bull (Bos taurus) exhibiting 23% macrocephalic spermatozoa in the ejaculate was investigated. Spermatozoa with a projected head area of ≥52 μm2 were considered macrocephalic. Diploidy was assumed from the measurement of sperm head area and proved by flow cytometry, which was used to sort the sperm into haploid and diploid fractions. Fluorescence in situ hybridization was used to detect the sex chromosomes with an X-Y probe set. Diploid spermatozoa most likely originate from a defective second meiotic division (M2 diploids), as only 0.7% XY-bearing spermatozoa (M1 diploids) were detected in the spermatozoa of the flow cytometric diploid sort. The painting probes generated a single X or Y spot for both unsorted semen and diploid sorted spermatozoa. This indicates a close proximity of the nonpartitioned sister chromatids in the spermatozoa. The BC1.2 probe, which labels BTAYp13-12, was used to clarify the presence of the two chromatids in the singular signal of the simultaneously hybridized Y-painting probe. In scoring more than 1000 randomly sampled spermatozoa hybridized with the BC1.2 probe, 32% showed the YY diploid signal and 18% the Y signal. The sperm diploidy in this bull was caused by an incomplete partitioning of sister chromatids during the second meiotic division (M2) associated with a failure in nuclear cleavage.