Tyrosine phosphorylated proteins have been localized and identified in male reproductive tissues such as testis and capacitated/ acrosome reacted sperm except epididymis. The changes of such proteins are associated with decreased sperm quality of valproic acid treatment. This study aimed to investigate the presence and alterations of protein phosphorylation in epididymal epithelium and fluid of rats treated VPA. Sixteen adult male rats were divided into control and VPA-treated groups (n=8/ each). Treated rats were injected with VPA (500 mg/ kgBW, intraperitoneally) for 10 consecutive days. At the end of experiment, the monoclonal antiphosphotyrosine (clone 4G10) was used for immunohistochemistry to probe tyrosine phosphorylated proteins and also to examine the expression of such proteins using immuno-Western blotting in epididymal tissue and fluid. The result showed that positive reactivity of phosphorylated proteins was clearly observed in cytoplasmic principle cells, nuclei of apical & basal cells and sperm mass surrounded with epididymal fluids. The profiles of phosphorylated proteins in epididymal fluid were 182, 127, 80, 70, 57, 45, 34, and 31 kDas, respectively. Interestingly, VPA affected the changes of phosphorylated proteins and β actin in head, body, and tail epididymal fluids. We conclude that tyrosine phosphorylated proteins were detected in epididymal epithelium and fluid. The expressions of those proteins and actin were altered under VPA treating.

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SITTHICHAI, I.; WANNISA, S.; APICHAKARN, S. & BUNGORN, S.Protective effect of Momordica cochinchinensis (L.) spreng aril extract on essential testicular markers in rats induced with valproic acid. Int. J. Morphol., 35(3):992-999, 2017. SUMMARY:Valproic acid (VPA), an anti-epileptic drug, has been reported to cause male sub/infertility. Together with searching for alternative treatments, the degrees to which testosterone levels and sperm quality are decreased under VPA treatment also need to be clarified. This study aimed to investigate the protective effects of Momordica cochinchinensis (MC) aril extract containing antioxidant capacity on adverse reproductive parameters induced with VPA. Rats were divided into 6 groups (control, VPA, 200 mg kg-1 of MC only, 50, 100, 200 mg kg-1 MC+VPA, respectively, n=8 in each). Animals were pretreated with MC extract for 23 days before coadministration with VPA (500 mg kg-1, i.p.) for 10 consecutive days. All reproductive parameters including histology, and expression of androgen receptor (AR), Ki-67, tyrosine phosphorylated proteins, and steroidogenic proteins in testis were examined. The results showed that MC could prevent all reproductive parameters in VPA-treated rats. Moreover, MC+VPA groups showed significant declining of testicular histopathologies compared to VPA group. It also decreased the malondialdehyde level and changes of the testicular StAR, AR, and tyrosine phosphorylated protein expressions. In conclusion, M. cochinchinensis aril extract can prevent adverse male reproductive parameters and essential testicular proteins damages induced with VPA.