Environmental temperature has effects on sperm quality with differences in susceptibility between cattle subspecies and breeds, but very little is known about the seminal plasma protein (SPP) changes resulting from testicular heat stress. Scrotal insulation (SI) for 48 hr was applied to Brahman (Bos indicus) bulls. Semen was collected at 3-day intervals from before, until 74 days post-SI. The changes in sperm morphology and motility following SI were comparable to previously reported and differences were detected in measures of sperm chromatin conformation as early as 8 days post-SI. New proteins spots, in the SPP two-dimensional (2-D) gels, were apparent when comparing pre-SI with 74 days post-SI, and SPP identified as associated with mechanisms of cellular repair and protection. Similar trends between 2-D gel and Sequential Window Acquisition of All Theoretical Mass Spectra (SWATH -MS) data was observed, with SWATH-MS able to quantify individual SPP that otherwise were not resolved on 2-D gel. The SPP assessment at peak sperm damage (21-24 days) showed a significant difference in 29 SPP (adjusted p < .05), and identified six proteins with change in abundance in the SI group. In conclusion both spermatozoa and SPP composition of bulls are susceptible to temperature change incurred by SI, and SPP markers for testicular heat insults may be detected.
K E Y W O R D Schromatin structure, MS-based proteomics, scrotal insulation, testicular stress Note: Period 1-impact on epididymal maturation (2-11 days post-SI), Period 2-impact on spermiogenesis and meiosis (14-41 days post-SI), Period 3-impact on spermatocytogenesis (44-62 days post-SI), and Period 4-impact after one full wave of spermatogenesis (65-74 days post-SI); where Day 0 represents initiation of the 48 hr period of SI. The data are presented as means ± SE for bulls with applied SI for 48 hr (n = 3) and control (n = 3). The PNS is further differentiated into the proportion of spermatozoa with HD, D, MPD, and TD.