Differential Expression and Localization of EHBP1L1 during the First Wave of Rat Spermatogenesis Suggest Its Involvement in Acrosome Biogenesis

The identification and characterization of new proteins involved in spermatogenesis is fundamental, considering that good-quality gametes are basic in ensuring proper reproduction. Here, we further analyzed the temporal and spatial localization during the first spermatogenic wave of rat testis of EHBP1L1, which is involved in vesicular trafficking due to the CH and bMERB domains, which bind to actin and Rab8/10, respectively. Western blot and immunofluorescence analyses showed that EHBP1L1 protein expression started at 21 days post-partum (dpp) concomitantly with the appearance of primary spermatocytes (I SPC). In subsequent stages, EHBP1L1 specifically localized together with actin in the perinuclear cytoplasm close to the acrosomal and Golgian regions of spermatids (SPT) during the different phases of acrosome biogenesis (AB). Moreover, it was completely absent in elongated SPT and in mature spermatozoa, suggesting that its role was completed in previous stages. The combined data, also supported by our previous report demonstrating that EHBP1L1 mRNA was expressed by primary (I) and secondary (II) SPC, lead us to hypothesize its specific role during AB. Although these results are suggestive, further studies are needed to better clarify the underlying molecular mechanisms of AB, with the aim to use EHBP1L1 as a potential new marker for spermatogenesis.

Effect of atorvastatin on spermatogenesis in rats: A stereological study

Purpose: To investigate the effects of oral atorvastatin on spermatogenesis in a rat model.Methods: Rats were equally assigned into control and study groups, the latter receiving atorvastatin (20 mg/kg/day). At the end of 12 weeks, spermatogenetic activity was evaluated using stereological and optical fractionator methods. Serum follicle-stimulating hormone (FSH), total testosterone (TT), and luteinizing hormone (LH) levels were measured using micro–ELISA kits. Total cholesterol, triglyceride (TG), low-density lipoprotein cholesterol (LDL - C), and high-density lipoprotein cholesterol levels were also measured by enzymatic colorimetric assays.Results: Testicular stereological analysis revealed that atorvastatin reduced Sertoli cell numbers (p < 0.001), spermatogonia (p < 0.001), spermatocytes (p < 0.001), and seminiferous tubule diameters (p < 0.001). LDL – C (p = 0.01) and TG (p = 0.01) values were significantly lower in the study group compared with the control group. There was no significant difference in FSH (p = 0.44), LH (p = 0.48),and TT (p = 0.06) levels between the groups.Conclusion: The findings show that atorvastatin causes deleterious effects on rat spermatogenesis. It should therefore be used with caution in clinical practice owing to its potential adverse effects, especially on male fertility. Keywords: Statin, Atorvastatin, Spermatogenesis, Stereology, Testis

Rat in vitro spermatogenesis promoted by chemical supplementations and oxygen-tension control

In vitro spermatogenesis (IVS) using air–liquid interphase organ culture method is possible with mouse testis tissues. The same method, however, has been hardly applicable to animals other than mice, only producing no or limited progression of spermatogenesis. In the present study, we challenged IVS of rats with modifications of culture medium, by supplementing chemical substances, including hormones, antioxidants, and lysophospholipids. In addition, reducing oxygen tension by placing tissues in an incubator of lower oxygen concentration and/or applying silicone cover ceiling on top of the tissue were effective for improving the spermatogenic efficiency. Through these modifications of the culture condition, rat spermatogenesis up to round spermatids was maintained over 70 days in the cultured tissue. Present results demonstrated a significant progress in rat IVS, revealing conditions commonly favorable for mice and rats as well as finding rat-specific optimizations. This is an important step towards successful IVS in many animal species, including humans.

Effect of exposure to active allethrin on expression of HSF2 and Ovol1 genes in a rat spermatogenesis

Background: Heat shock factor protein 2 (HSF 2) and Ovo like transcriptional repressor 1 (Ovol1) genes are found in germ cells that control spermatogenesis. One reason is exposure to mosquito repellent drugs from allethrin. Allethrin is one of the causes of male reproductive dysfunction that affects male reproduction resulting in infertility.Methods: This study was an experimental post-test randomized control group design. Twenty-eight rats given exposure according to the experimental group were K (control), P1 (4 hours exposure), P2 (8hours exposure), and P3 (12hours exposure) for 30 days. Examination of HSF 2 and Ovol1 genes from testicular tissue using real-time PCR with a relatively quantitative calculation method. Implementation of research at animal houses and biomedical laboratories. Data analysis using one-way ANOVA with a significant level of p <0.05.Results: The results of this study found differences in the average number of expressions of the HSF2 and Ovol1 genes. The average expression of the HSF2 control group gene was 3.50, P1: 2.99, P2: 0.62, and P3: 0.49. Whereas in the Ovol1 gene control group were 1.17, P1: 0.80, P2: 0.57, and P3: 0.65. The results of statistical tests using one-way ANOVA are HSF2 (p = 0,000) and Ovol1 (p = 0.045).Conclusions: Based on the results of this study, it was concluded that there was an allethrin effect in decreasing the expression of the HSF 2 and Ovol1 genes in Wistar albino Rattus Novergicus strain.