Sperm cryopreservation is an important method of assisted reproductive techniques and storing genetic resources. It plays a vital role in genetic improvement, livestock industrial preservation of endangered species, and clinical practice. Consequently, the cryopreservation technique is well organized through various studies, especially on Korean native cattle (Hanwoo). However, the cryopreservation technique of Korean native brindled cattle, which is one of the native cattle species in Korea, is not well organized. Therefore, it is necessary to develop a Supplementary Table technique for the cryopreservation of Korean native brindled cattle. For this purpose, it is important to first evaluate the quality of the currently produced cryopreserved sperm of Korean native brindled cattle. In this study, we randomly selected 72 individual Korean native brindled cattle semen samples collected from 8 different region research centers and used them to evaluate sperm functions. We focused on the quality evaluation of cryopreserved Korean native brindled cattle semen following the measurement of motion kinematics, capacitation status, intracellular ATP level, sperm motility, and cell viability. Then, the values of each of the eight groups were derived from various sperm parameters of nine individual samples, including sperm motility, kinematics, cellular motility, and intracellular ATP levels, which were used to compare and evaluate sperm function. Overall, differences in various sperm parameters were observed between most of the research centers. Particularly, the deviations of motility and motion kinematics were high according to the sample. Therefore, we suggest that it is necessary to develop a standard method for the cryopreservation of Korean native brindled cattle semen. We also suggest the need for sperm quality evaluation of the cryopreserved semen of Korean native brindled cattle before using artificial insemination to attain a high fertility rate.

As an endocrine disruptor, bisphenol-A (BPA) causes several functional and behavioral abnormalities related to reproduction. The current study was design to evaluate the effect of perinatal exposure of female mice to BPA on sperm function of adult F(1) offspring. Pregnant female mice F(0) were gavaged with three different concentration of BPA, such as 50 μg/kg/day (tolerable daily intake value by the European Food Safety Authority), 5 mg/kg/day (no-observed-adverse-effect level; NOAEL), and 50 mg/kg/day (lowest-observed-adverse-effect level; LOAEL) and corn oil (7 mg/kg/day; vehicle control). The functional parameters of F(1) spermatozoa were studied both before and after capacitation, whereas the fertility assessment was evaluated by in vitro and in vivo assay using unexposed females. Our results showed that spermatozoa hyperactivated motility, capacitation, intracellular ATP, Ca2+, and ROS levels after capacitation were significantly affected using NOAEL and LOAEL concentration of BPA. However, the sperm motility was only affected by LOAEL dose after capacitation. All of the tested parameters were potentially unaffected by BPA before capacitation, except intracellular ATP that decreased by all concentrations. Although both NOAEL and LOAEL concentration were effectively reduced the rate of fertilization and embryonic development in vitro, however the average litter size was only affected by LOAEL dose. Our finding suggested that perinatal exposure of 50 μg/kg/day did not produce significant effects; however both NOAEL and LOAEL affects overall sperm function after capacitation, leading to impairments in the fertility of F(1) male offspring.

After vasectomy (VAX), the flux and composition of the epididymal fluid are modified, leading to sequelae to the epididymis. In an effort to understand molecular pathophysiology of the epididymis following VAX, we investigated the changes of gene expression and sperm functions in the epididymis of vasectomized mice. After VAX, the epithelial cell height was significantly decreased in cauda epididymis, resembling the those of vas deference. This suggests that VAX evokes alteration of segmental characteristics of epididymal epithelium. Of note, these was an increase in luminal diameter in corpus and cauda epididymis, indicating the alteration of fluid homeostasis in epididymal lumen and protein synthesis and secretion. Also, the formation of sperm granuloma and infiltration of inflammatory cells were noted in lumen of epididymis at 8 weeks postvasectomy, indicating the activation of immune response in epididymis. The serum TNF-α levels and epididymal TNF-α and IL-1β mRNA levels were significantly increased in VAX mice. Microarray analysis demonstrated that claudin (CLDN) 10 and cystic fibrosis transmembrane conductance regulator (CFTR) were downregulated after vasectomy. In contrast, angiotensin II receptor type 2 (Agtr2) was up-regulated after vasectomy. Taking into account the functional importance of angiotensin system in epididymal epithelia and muscle tissue, VAX may alter the secretory function of epithelia and sperm transport via alteration in angiotensin system. Importantly, the IgG type of anti-sperm antibody (ASA) were markedly increased in the blood of vasectomized mice. Together this indicates that VAX provokes local inflammation in epididymis as well as systemic inflammation, which in turn change the blood epididymal barrier, an important element for epididymal immunological privilege. In addition, the number of sperm in cauda epididymis was increased but the sperm motility was decreased after vasectomy. The spontaneous acrosome reaction was increased by vasectomy after capacitation. This suggests that VAX affects sperm functions as well as sperm maturation. In conclusion, bilateral vasectomy lead to local immune response of epididymis, causing immunologic infertility in men.