Prolonged epididymal sperm storage in vespertilionid and rhinolophid bats, provides an interesting experimental model for the study of spermatozoa epididymal maturation. We examined the presence of the cytoplasmic droplet, and the sequential induction of capacitation and the acrosome reaction in spermatozoa obtained from different epididymal regions (caput, corpus, cauda) throughout the annual reproductive cycle of Corynorhinus mexicanus (C. mexicanus). This is a vespertilionid bat that stores spermatozoa in the epididymis for several months after testes regression. The number of sperm recovered from the different epididymal regions indicate that epididymal transit in C. mexicanus is rapid. The persistence of a high percentage of sperm cells with cytoplasmic droplet in cauda epididymis was observed in addition to a low index of capacitation and acrosome reaction in sperm cells obtained from the corpus epididymis. There was a significant increase in the percentage of capacitated and acrosome reacted spermatozoa during the storage of sperm cells in the cauda epididymis and the percentage of capacitated spermatozoa was consistently, and significantly, higher (p < 0.05) in cauda compared to the corpus epididymis at all studied dates. Tthe process of epididymal maturation in C. mexicanus is completed in the caudal region of this organ encompassing a significant period. Our results also indicate that in C. mexicanus, and in other vespertilionid and rhinolophid bats that show the same temporal asynchrony in the function of male reproductive organs, the final phases of epididymal maturation and storage are, apparently, independent of testicular function.
The bat Corynorhinus mexicanus provides an interesting experimental model for the study of epididymal sperm maturation because after spermatogenesis and the regression of the testes, this bat stores sperm in the epididymal cauda for several months. Earlier research conducted by our group suggested that sperm maturation in this species must be completed in the caudal region of the epididymis. One of the major signal transduction events during sperm maturation is the tyrosine phosphorylation of sperm proteins. The aim of the present study was to comparatively evaluate tyrosine phosphorylation in spermatozoa obtained from the caput, corpus and cauda of the epididymis during the sperm storage period. The maturation status of the sperm was determined by the percentage of capacitation and tyrosine phosphorylation in sperm obtained from the epididymis. The highest proportion of tyrosine phosphorylation was registered after the sperm had reached the cauda epididymis during the middle of the storage period. In conclusion, in Corynorhinus mexicanus and most likely in other chiropteran species with an asynchronous male reproductive pattern, epididymal sperm maturation ends in the caudal region of the epididymis and is related to the time that the sperm remains in the epididymis before mating activity.
Stress is associated with detrimental effects on male reproductive function. It is known that stress increases reactive oxygen species (ROS) generation in the male reproductive tract. High ROS levels may be linked to low sperm quality and male infertility. However, it is still not clear if ROS are generated by stress in the testis. The objective of this study was to characterize the role of oxidative stress induced by cold-water immersion stress in the testis of adult male rats and its relation with alterations in cauda epididymal sperm. Adult male rats were exposed to acute stress or chronic stress by cold-water immersion. Rats were sacrificed at 0, 6, 12, and 24 hours immediately following acute stress exposure, and after 20, 40, and 50 days of chronic stress. ROS production increased only at 6 hours post-stress, while the activity and expression of antioxidant enzymes, lipid peroxidation (LPO), and sperm parameters were not modified in the testis. Corticosterone increased immediately after acute stress, whereas testosterone was not modified. After chronic stress, testicular absolute weight decreased; in addition, ROS production and LPO increased at 20, 40, and 50 days. The activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx) decreased throughout the duration of chronic stress and the activity of catalase (CAT) decreased at 40 and 50 days, and increased at 20 days. The expression of copper/zinc superoxide dismutase (SOD1) and CAT were not modified, but the expression of phospholipid hydroperoxide glutathione peroxidase (GPx-4) decreased at 20 days. Motility, viability, and sperm count decreased, while abnormal sperm increased with chronic stress. These results suggest that during acute stress there is a redox state regulation in the testis since no deleterious effect was observed. In contrast, equilibrium redox is lost during chronic stress, with low enzyme activity but without modifying their expression. In addition, corticosterone increased while testosterone decreased, this decrease is related to the negative effects seen in sperm.
We studied the activities of reactive oxygen species (ROS) scavenging enzymes during epididymal spermatozoon maturation and storage in Corynorhinus mexicanus (G.M. Allen, 1916), a vespertilionid bat that stores spermatozoa in the epididymides for several months after regression of the testes. Depending on the phase of the epididymal reproductive cycle, two different patterns of antioxidant enzyme activities were observed in C. mexicanus. Catalase activity is clearly present in both caput and cauda epididymides throughout the entire annual reproductive cycle, being particularly high during the post-testicular phase of epididymal function. Superoxide dismutase (SOD) activity, present during the testicular phase of epididymal transport and maturation of spermatozoa, is almost completely absent or inhibited in both epididymal segments during the post-testicular epididymal storage period. GPx activity is low during the testicular phase of epididymal spermatozoon maturation and is high in both epididymal segments during the storage phase of epididymal function. From our results, we postulate that (i) the pattern of epididymal antioxidant enzyme activities in C. mexicanus is significantly different from the pattern that is proposed to be unique for mammals; (ii) epididymal function in these species of bats can be clearly divided into two phases, a testicular-dependent phase that is related to the spermatozoon maturation function of the epididymides and a testicular-independent phase that is related to the long-term spermatozoon storage function observed in these mammals; (iii) the study of the regulation of the redox potential of the microenvironment, associated with mammalian spermatozoa as they transit through the epididymides, must be particularly focused on the anatomical region where ROS generation scavenging and spermatozoon maturation storage processes take place.
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