Since flutamide treatment and aging also affect spermatogenesis and sperm motility, it should be noted that epididymal sperm donors differ from seminal vesicle donors (Figure 1H). The results showed that motile, progressive motile, LIN, and VSL were significantly reduced in the sperm cultured in seminal vesicle secretions from flutamide-treated (Flutamide) compared to the sperm cultured in seminal vesicle secretions from controls (Ctrl) (Figure 1I–L). Note that the donor mice for sperm and seminal vesicle secretions were different. (A) The effects of secretions from the prostate and seminal vesicles on sperm motility were directly compared. These results indicate that sperm exposure to seminal plasma factors significantly improved sperm linear motility (high VSL and LIN). When seminal vesicle secretions were added to the human tubal fluid (HTF) medium, the sperm motility, progressive motility, the straight line velocity (VSL), and the percentage of linearity (LIN) values increased significantly compared to when only prostate extract was used. A direct comparison was conducted to clarify the effects of secretions from the prostate and seminal vesicles on the motility parameters of epididymal sperm. This important work elucidates the biological processes and detailed mechanisms by which testosterone influences seminal plasma metabolites in mice. The contaminating cell type was considered to be mainly muscle cells because the gene expression levels of muscle cell markers verified by RNA-seq were relatively high. The seminal vesicle secretions from flutamide-treated mice were also collected similarly. Low levels limit these processes, causing muscle mass to decline over time. When levels fall, the body struggles to generate energy efficiently. Testosterone influences metabolism and red blood cell production. Without adequate testosterone, this pathway weakens, making erections difficult to achieve or maintain. Though testosterone is not the only hormone responsible for erections, it is crucial. This is because testosterone acts directly on the brain's limbic system, which governs arousal and sexual motivation. If you're experiencing these symptoms, it may be time to check your hormone levels. Conceptualization, Data curation, Formal analysis, Investigation, Visualization, Methodology, Writing – original draft, Writing – review and editing. The funders had no role in study design, data collection, and interpretation, or the decision to submit the work for publication. All statistical details of the experiments are given in the figure legends. The full-length blotting images are shown in Figure 6 and Figure 7—source data 2 or 3. The membranes were then treated with Tris-buffered saline and Tween 20 (TBST, 20 mM Tris, pH 7.5, 150 mM NaCl, 0.1% Tween 20) containing 5% (wt/vol) Non-Fat Dry Milk (MORINAGA MILK INDUSTRY Co, Ltd, Tokyo, Japan) to block nonspecific reactions. Sperm motility patterns were subsequently examined using CASA. It is known that intratesticular T can be reduced substantially without an effect on spermatogenesis (5). Indeed, the ratios for the 2 proteins among hypogonadics and controls, obtained from Western blot data (ratio 0.41 and 0.77, respectively), were very similar to the ones observed by proteomics data (ratio 0.51 and 0.74, respectively) (Figures 2B, C; Table 2). Sperm protein extracts from 4 out of the 5 hypogonadic patients used for the proteomic analysis, and 4 out of the 5 fertile controls, were used for the Western blot analysis (two samples were completely used for proteomic analysis). Worth mentioning, some of these proteins had more than one main cellular function, and the 43 proteins were involved in 71 cellular functions. Results are expressed as the protein ratio of sperm proteins from secondary hypogonadism patients (Hypo) to controls (Ctl). Because seminal plasma contains components specific to certain male reproductive organs, differences in protein composition may reflect pathological processes in these specific organs (59). Cell hypertrophy is partly due to the accumulation of fatty acids in the cytoplasm, which causes significant damage to cellular functions (46, 47). Manipulation of ACLY expression in mouse myoblasts and embryonic stem cells has been shown to alter the TCA cycle into a "non-normal cycle" (42). On the other hand, fructose and lipids in seminal plasma are the major energy sources for sperm (35). Mammalian seminal plasma is secreted mainly from the seminal vesicle and is rich in cytokines, prostaglandins, and SVS family members (34). Chronic anxiety or tension can diminish sperm count and sexual function. High stress elevates cortisol, which suppresses testosterone production by interfering with the HPG axis. Antioxidants fight oxidative stress in the testes, protecting sperm DNA and quality. Clinical studies show it can effectively raise testosterone levels without compromising fertility. For men concerned about both low testosterone and male infertility, there are several evidence-backed options. For men who want to maintain fertility, alternatives like clomiphene or hCG injections may help stimulate the body's natural testosterone and sperm production without shutting it down. However, it's important to understand that TRT can reduce fertility by suppressing the body's natural production of testosterone and sperm. This is due to reduced activity in Leydig cells in the testes, which produce testosterone. Even if libido and erection function seem normal, poor sperm health can cause male infertility. How do you know if your testosterone levels might be impacting your fertility?