ERO1α promotes testosterone secretion in hCG-stimulated mouse Leydig cells via activation of the PI3K/AKT/mTOR signaling pathway

ER oxidoreduclin 1α (ERO1α) is an oxidase, participating in formation of secretory and membrane proteins. However, the other physiological functions ERO1α is not well known. We found that ERO1α is high in the Leydig cells of the testis. Therefore, the purposes of the current study are to explore the role of ERO1α and the possible mechanisms in regulating cell proliferation, apoptosis, and testosterone secretion of Leydig cells. ERO1α was mainly localized in Leydig cells in the adult mice testes by immunofluorescence staining. Western blot analysis showed that ERO1α was higher in Leydig cells than that in the seminiferous tubules. The effect of ERO1α on cell proliferation, apoptosis, and testosterone secretion was detected by transducing ERO1α overexpression and knockdown lentiviruses into cultured primary Leydig cells (PLCs) together with hCG exposure. Flow cytometry analysis showed that ERO1α promoted cell proliferation by increasing cell distribution at the S phase and decreasing that at the G0/G1 phase. Western bolt analysis showed that ERO1α increased CDK2 and CDK6 expression. Cell apoptosis determination found that ERO1α inhibited PLC apoptosis. Western bolt analysis showed that ERO1α increased the ratio of BCL-2/BAX, and decreased BAD and Caspase-3 expression. Enzyme-linked immunosorbent assay analysis demonstrated that ERO1α enhanced testosterone secretion. Western bolt analysis found that ERO1α increased StAR, 3β-HSD, and CYP17A1 expression. Furthermore, ERO1α could activate the PI3K/AKT/mTOR signaling pathway. In summary, these results suggest that ERO1α might play proliferation promotion and antiapoptotic roles and enhance testosterone secretion in PLC, at least partly, via activation of the PI3K/AKT/mTOR signaling pathway.     

Reproductive exocrine and endocrine profiles and their seasonality in male langur monkeys (Presbytis entellus entellus)

The reproductive exocrine and endocrine profiles in male langurs are reported with an emphasis on seasonality. The animals showed positive response to electroejaculation throughout the year. The sperm concentration varied between 10–383 × 10⁶/ejaculation with wide fluctuations all through the year. No appreciable changes in the motility and percent live sperm were observed throughout the year. The levels of seminal fructose and magnesium remained unchanged throughout the year, while acid phosphatase showed wide fluctuations. Citric acid showed elevation during February and March and LDH showed elevated levels during April and May. The annual range of serum testosterone was 6–34 nMol/l with a peak during July. Cortisol ranged between 575–1587 nMol/l and prolactin ranged between 107–900 mU/l. Wide fluctuations were observed in hormonal levels. No seasonality was exhibited in the seminiferous tubule diameter, nuclear diameter of Sertoli cells and Leydig cells, and the cholesterol, glycogen, and sialic acid contents of testis. None of the parameters studied have shown any correlation with season. The results suggest that the male langurs lack seasonality in their reproductive exocrine and endocrine profiles and thus could be used as model for research in human reproduction.     

An automated assay for fecal estrogen conjugates in the determination of sex in avian species

Recently captive propagation of psitticines has become necessary in order to preserve and breed endangered species. As habitat destruction increases and importation restrictions become more stringent, the need for a reliable and noninvasive technique for sex assignment is essential to improve the productivity of breeding and raising birds in captivity. One aspect of breeding is the proper assignment of breeding pairs; however, sex identification can be challenging due to the lack of sexual dimorphism in approximately 30% of all avian species. Historically, visual, behavioral, surgical, cytogenetic, and endocrine methods have been utilized in sex identification of avian species; however, the practicality, safety, and reliability of these techniques have been questioned. Therefore, there remains a need for an accurate, simple, and non-invasive method to identify the sex of monomorphic birds so that the formation of breeding pairs and the success of breeding programs is facilitated. The data in this study support the concept that an automated fluorescence-immunoassay which measures fecal estrogen conjugates can have an 87% success rate in determining the sex of psitticines, as demonstrated with cockatiels, while offering the advantages of non-invasiveness, simplicity, and speed.     

Reduction of 4-androstene-3,17-dione by growing cucumber plants

4-[4-¹⁴C]Androstene-3,17-dione was applied to the leaves of growing cucumber plants, Cucumis sativus, twice a week. Four weeks after the first     

A review of hormonal factors influencing the sexual and aggressive behavior of macaques

The effects of gonadal hormones on the sexual and aggressive behavior of adult macaques are reviewed. Similarities among findings from field, colony, and laboratory studies strengthen the view that testosterone facilitates the sexual and aggressive behavior of males, while sexual and perhaps aggressive behavior by the female is mainly dependent on estradiol, which increases both the sexual motivation of the female and her attractiveness to males. Differences between results from different settings help to emphasize the role of environmental and social factors in modulating the effects of hormones. © 1993 Wiley-Liss, Inc.     

Cold-induced phase separation for the simple and reliable extraction of sex hormones for subsequent LC-MS/MS analysis

Sex hormones, including androgens, estrogens, and progestogens, are important biomarkers for various diseases. Quantification of sex hormones is typically conducted by LC-MS/MS. At present, most methods require liquid-liquid extraction or solid phase extraction for sample preparation. However, these pretreatments are prone to compromise LC-MS/MS throughput. To improve on the current standard practices, we investigated cold-induced phase separation for sex hormone extraction. After protein precipitation with acetonitrile and adjusting the solution constitution with water, samples were stored at ?30°C for 10 min to generate two distinct phases: an acetonitrile-rich layer on top of a water-rich layer. During this process, the hydrophobic sex hormones spontaneously separate into the upper layer. This simple and reliable cold-induced phase separation-based LC-MS/MS methodology was used here to simultaneously detect estrone, estradiol, estriol, testosterone, androstenedione, dehydroepiandrosterone, progesterone, and 17-hydroxyprogesterone in serum. Validation of this method indicated satisfactory performance, including acceptable linearity, accuracy, precision, and tractability. Compared with the mainstream liquid-liquid extraction-based method, this new method exhibits significant progress in throughput, which shortens the time cost of sample preparation from 90 to 40 min. We propose that this method can be an excellent alternative for sex hormone analysis in routine clinical laboratories.     

The Effect of Testosterone Upon the Urate Reabsorptive Transport System in Mouse Kidney

It is hypothesized that hyperuricemia in males is caused by androgen-induced urate reabsorptive transport system in the kidney. The expression of urate transporter 1 (Urat1), sodium-coupled monocarboxylate transporter 1 (Smct1) and glucose transporter 9 (Glut9) were investigated in orchiectomized mice with or without testosterone replacement. Testosterone enhanced mRNA and protein levels of Smct1 while those of Glut9 were attenuated. Although the mRNA level of Urat1 was enhanced by testosterone, the corresponding levels of Urat1 protein remained unaffected. Thus, the induction of Smct1 by testosterone is a candidate mechanism underlying hyperuricemia in males.     

Effects of neonatal surgical castration and immunocastration in male pigs on blood T lymphocytes and health markers

Surgical castration in pig husbandry is criticized for welfare reasons. Thus, it is necessary to evaluate alternative ways of rearing male pigs, such as entire or immunocastrated animals. Immunocastration is a vaccination directed against gonadotropin-releasing hormone (GnRH) to suppress the production of sexual hormones. This study aimed at investigating the effects of these two methods of castration in comparison with intact male pigs on blood T-lymphocyte subsets and function, the immunoglobulin (Ig) response to an influenza vaccine and health markers during sexual development. A total of 70 animals were allocated to three experimental groups: entire (E), surgically castrated at 5 to 6 days of age (SC), and immunized against GnRH at 3 and 4 months of age (IC). Blood samples were collected at 3, 4 and 5 months. At slaughter, global health status and body and spleen weights were measured. Results showed that SC male pigs had fewer blood lymphocytes than E pigs at 4 and 5 months (P<0.05), whereas IC pigs did not differ significantly from E pigs. The percentages of CD3⁺, CD3⁺CD4⁺ and CD3⁺CD8⁺ lymphocytes were not altered by treatment (P>0.1). Compared with E pigs, the SC pigs had a higher percentage of CD3⁺CD4⁺CD8⁺ cells at 4 months, whereas the IC pigs had a higher percentage at 5 months (P<0.05). Regarding γδT cells, SC pigs had a lower percentage than E pigs at 4 and 5 months (P<0.05), whereas IC pigs did not differ significantly from E pigs at any age. However, there were no consequences on T-lymphocyte proliferation and total IgG or anti-influenza Ig. At slaughter, relative spleen weight was decreased in IC pigs, whereas pneumonia score was decreased in SC pigs relatively to E pigs. Overall, no clear functional consequences of either method on commercial pig immune abilities were demonstrated, but more investigations are required to ascertain this conclusion.