(A) Western blot images of ACLY and α/β-tubulin in three sets of seminal vesicle epithelial cells cultured with 100 ng/ml testosterone (Testo) or in vehicle (Ctrl) for 7 days. The addition of testosterone significantly increased the amount of ACLY protein in seminal vesicle epithelial cells (Figure 7A, B). (D) Western blot images of GLUT4 and α/β-tubulin in three sets of seminal vesicle epithelial cells cultured with 100 ng/ml testosterone (Testo) or in vehicle (Ctrl) for 7 days. In fact, in 2014 we published the first experimental proteomic study, using high-resolution mass spectrometry, aimed at studying the seminal proteome of patients affected by secondary hypogonadism, before and after 6 months of testosterone replacement treatment (26). In order to study if the reduction of serum T, although mild, might modify the qualitative protein composition of spermatozoa, we selected five patients affected by hypotestosteronemia due to isolated LH deficiency. The maintenance of spermatogenesis in humans requires an adequate secretion of LH, resulting in the maintenance of high intra-testicular testosterone (ITT) (2). Western Blot analysis for the PIP and AKAP3 proteins obtained from sperm samples of patients with secondary hypogonadism (HYPO) and controls (CTL) (A). The objective of this study was to elucidate the mechanisms of seminal plasma synthesis. Simultaneously, cell proliferation in the epididymal epithelial cells is suppressed. Although our findings and previous studies suggest that oleic acid may play an important role in improving fertility, it is still too early to draw a definitive conclusion from this study. It could be assumed that a decrease in testosterone levels causes abnormalities in the components of human semen. This indicates that testosterone plays a major role not only in spermatogenesis but also in the functional maturation of sperm. Although there is a commonality in the activation of the glycolytic system in testosterone target tissues, the different expression patterns of genes involved in metabolism in mitochondria may exert their specific functions in each tissue. To investigate the cause of this multilayering of cells, we performed Ki67 staining, a marker of cell proliferation, and apoptosis detection by TUNEL staining (Fig.2B). Each replicate experiments with 3-6 wells containing pooled cells from 3-5 mice. The epithelial cells were culutured with or without testosterone for 8 days. Manipulation of ACLY expression in mouse myoblasts and embryonic stem cells has been shown to alter the TCA cycle into a ‘non-normal cycle’ (Arnold et al., 2022). On the other hand, fructose and lipids in seminal plasma are the major energy sources for sperm (Lenzi et al., 1996). For this reason, recent studies have focused on the immunomodulatory function of seminal plasma in the female reproductive tract. Mammalian seminal plasma is secreted mainly from the seminal vesicle and is rich in cytokines, prostaglandins, and SVS family members (Gonzales, 2001). Additionally, while testosterone also significantly increased fatty acid synthesis, this increase was completely abolished by the GLUT4 inhibitor (Figure 8J). (G) The impact of ACLY knockdown on testosterone-induced fatty acid synthesis was measured.
