Objective: To study the effects of cadmium chloride (CdCl(2)) exposure on testicular autophagy levels and blood-testis barrier integrity in prepubertal male SD rats and testicular sertoli (TM4) cells. Methods: In July 2021, 9 4-week-old male SD rats were randomly divided into 3 groups: control group (normal saline), low dose group (1 mg/kg·bw CdCl(2)) and high dose group (2 mg/kg·bw CdCl(2)), and were exposed with CdCl(2) by intrabitoneal injection. 24 h later, HE staining was used to observe the morphological changes of testis of rats, biological tracer was used to observe the integrity of blood-testis barrier, and the expression levels of microtubule-associated protein light chain 3 (LC3) -Ⅰ and LC3-Ⅱ in testicular tissue were detected. TM4 cells were treated with 0, 2.5, 5.0 and 10.0 μmol/L CdCl(2) for 24 h to detect the toxic effect of cadmium. The cells were divided into blank group (no exposure), exposure group (10.0 μmol/L CdCl(2)), experimental group[10.0 μmol/L CdCl(2)+60.0 μmol/L 3-methyladenine (3-MA) ] and inhibitor group (60.0 μmol/L 3-MA). After 24 h of treatment, Western blot analysis was used to detect the expression levels of LC3-Ⅱ, ubiquitin binding protein p62, tight junction protein ZO-1 and adhesion junction protein N-cadherin. Results: The morphology and structure of testicular tissue in the high dose group were obvious changed, including uneven distribution of seminiferous tubules, irregular shape, thinning of seminiferous epithelium, loose structure, disordered arrangement of cells, abnormal deep staining of nuclei and vacuoles of Sertoli cells. The results of biological tracer method showed that the integrity of blood-testis barrier was damaged in the low and high dose group. Western blot results showed that compared with control group, the expression levels of LC3-Ⅱ in testicular tissue of rats in low and high dose groups were increased, the differences were statistically significant (P<0.05). Compared with the 0 μmol/L, after exposure to 5.0, 10.0 μmol/L CdCl(2), the expression levels of ZO-1 and N-cadherin in TM4 cells were significantly decreased, and the expression level of p62 and LC3-Ⅱ/LC3-Ⅰ were significantly increased, the differences were statistically significant (P<0.05). Compared with the exposure group, the relative expression level of p62 and LC3-Ⅱ/LC3-Ⅰ in TM4 cells of the experimental group were significantly decreased, while the relative expression levels of ZO-1 and N-cadherin were significantly increased, the differences were statistically significant (P<0.05) . Conclusion: The mechanism of the toxic effect of cadmium on the reproductive system of male SD rats may be related to the effect of the autophagy level of testicular tissue and the destruction of the blood-testis barrier integrity.
Rats ; Male ; Animals ; Testis ; Cadmium Chloride/metabolism* ; Cadmium ; Blood-Testis Barrier/metabolism* ; Rats, Sprague-Dawley ; Cadherins/metabolism* ; Autophagy

OBJECTIVETo explore the changes in the expressions of the tight junction related protein occludin and junctional adhesion molecule-1 (JAM-1) of the blood-testis barrier and their significance in rats after microwave radiation.

METHODSEighty male Wistar rats were exposed to microwave radiation with average power density of 0, 10, 30 and 100 mW/cm2 for five minutes, and dynamic changes in the expressions of testicular occludin and JAM-1 were observed by Western blot and image analysis at 6 h, 1 d, 3 d, 7 d and 14 d after the radiation.

RESULTSThere was a significant down-regulation in the expression of the occludin protein at 3 - 7 d, 6 h - 7 d and 6 h - 14 d (P < 0. 05), as well as in that of JAM-1 at 3 - 7 d, 1 - 7 d and 1-14 d (P < 0.05) after exposure to 10, 30 and 100 mW/cm2 microwave radiation.

CONCLUSIONThe decreased protein expressions of occludin and JAM-1 may play an important role in the microwave radiation induced-damage to the blood-testis barrier.

Animals ; Blood-Testis Barrier ; metabolism ; radiation effects ; Cell Adhesion Molecules ; metabolism ; Down-Regulation ; Male ; Membrane Proteins ; metabolism ; Microwaves ; Occludin ; Rats ; Rats, Wistar ; Testis ; metabolism ; radiation effects

Although spermatozoa are formed during spermatogenesis in the testis, testicular spermatozoa are immature and cannot swim or fertilize. These critical spermatozoal functions are acquired in the epididymis where a specific luminal environment is created by the blood-epididymal barrier; proteins secreted by epididymal principal cells bind to maturing spermatozoa and regulate the maturational process of the spermatozoa. In the epididymis, epithelial cell-cell interactions are mediated by adhering junctions, necessary for cell adhesion, and by tight junctions, which form the blood-epididymal barrier. The regulation of these cellular junctions is thought to represent a key determinant in the process of sperm maturation within the epididymis. Tight junctions between adjacent principal cells permit the formation of a specific microenvironment in the lumen of the epididymis that is essential for sperm maturation. Although we have made significant progress in understanding epididymal function and the blood-epididymal barrier, using animal models, there is limited information on the human epididymis. If we are to understand the normal and pathological conditions attributable to human epididymal function, we must clearly establish the physiological, cellular and molecular regulation of the human epididymis, develop tools to characterize these functions and develop clinical strategies that will use epididymal functions to improve treatment of infertility.
Animals ; Blood-Testis Barrier ; physiology ; Cadherins ; metabolism ; Cell Adhesion ; Epididymis ; blood supply ; physiology ; Humans ; Male ; Membrane Proteins ; metabolism ; Occludin ; Rats ; Spermatozoa ; physiology ; Tight Junctions ; physiology

OBJECTIVETo study the effect of electromagnetic pulse (EMP) exposure on the permeability of blood-testicle barrier (BTB) in mice.

METHODSAdult male BALB/c mice were exposed to EMP at 200 kV/m for 200 pulses with 2 seconds interval. The mice were injected with 2% Evans Blue solution through caudal vein at different time points after exposure, and the permeability of BTB was monitored using a fluorescence microscope. The testis sample for the transmission electron microscopy was prepared at 2 h after EMP exposure. The permeability of BTB in mice was observed by using Evans Blue tracer and lanthanum nitrate tracer.

RESULTSAfter exposure, cloudy Evans Blue was found in the testicle convoluted seminiferous tubule of mice. Lanthanum nitrate was observed not only between testicle spermatogonia near seminiferous tubule wall and sertoli cells, but also between sertoli cells and primary spermatocyte or secondary spermatocyte. In contrast, lanthanum nitrate in control group was only found in the testicle sertoli cells between seminiferous tubule and near seminiferous tubule wall.

CONCLUSIONEMP exposure could increase the permeability of BTB in the mice.

Animals ; Blood-Testis Barrier ; metabolism ; radiation effects ; Coloring Agents ; Electromagnetic Fields ; Evans Blue ; Lanthanum ; Male ; Mice ; Mice, Inbred BALB C ; Permeability ; radiation effects ; Seminiferous Tubules ; metabolism ; radiation effects

OBJECTIVETo explore the effects of long-term exposure to particulate matter 2.5 (PM2.5) from automobile exhaust on the reproductive function of Sprague Dawley (SD) rats.

METHODSForty-five male SD rats, weighing 80 - 94 g and aged 28 days, were randomly assigned to receive intra-tracheal administration of 0.9% normal saline (control group, n = 15), PM2. 5 at 2 μg per 100 g body weight per day (low-dose PM2.5 group, n = 15), and PM2.5 at 16 μg per 100 g body weight per day (high-dose PM2.5 group, n = 15), qd, for 60 successive days. After the last 24-hour exposure, 10 rats were taken from each group for copulation with normal female ones, while the others were sacrificed, their testes removed for sperm count and deformity, pathological examination, and determination of the Connexin43 expression.

RESULTSThe conception rate was significantly decreased in the low- and high-dose PM2.5 groups as compared with that of the control (70% and 50% vs 100%), and so were the sperm count and quality. The rats in the PM2.5-exposed groups showed significantly disordered histological structure of the seminiferous tubules, reduced sperm count in the testicular lumen, some exfoliated secondary spermatocytes, downregulated Connexin43 expression in the testis, and damaged blood-testis barrier.

CONCLUSIONLong-term exposure to PM2.5 from automobile exhaust damages the reproductive function of male SD rats.

Animals ; Blood-Testis Barrier ; Body Weight ; Connexin 43 ; metabolism ; Down-Regulation ; Fertilization ; Male ; Particulate Matter ; toxicity ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reproduction ; Seminiferous Tubules ; Sperm Count ; Spermatocytes ; Testis ; metabolism ; pathology ; Vehicle Emissions ; toxicity

During spermatogenesis, developing germ cells that lack the cellular ultrastructures of filopodia and lamellipodia generally found in migrating cells, such as macrophages and fibroblasts, rely on Sertoli cells to support their transport across the seminiferous epithelium. These include the transport of preleptotene spermatocytes across the blood-testis barrier (BTB), but also the transport of germ cells, in particular developing haploid spermatids, across the seminiferous epithelium, that is to and away from the tubule lumen, depending on the stages of the epithelial cycle. On the other hand, cell junctions at the Sertoli cell-cell and Sertoli-germ cell interface also undergo rapid remodeling, involving disassembly and reassembly of cell junctions, which, in turn, are supported by actin- and microtubule-based cytoskeletal remodeling. Interestingly, the underlying mechanism(s) and the involving biomolecule(s) that regulate or support cytoskeletal remodeling remain largely unknown. Herein, we used an in vitro model of primary Sertoli cell cultures that mimicked the Sertoli BTB in vivo overexpressed with the ribosomal protein S6 (rpS6, the downstream signaling protein of mammalian target of rapamycin complex 1 [mTORC1]) cloned into the mammalian expression vector pCI-neo, namely, quadruple phosphomimetic and constitutively active mutant of rpS6 (pCI-neo/p-rpS6-MT) versus pCI-neo/rpS6-WT (wild-type) and empty vector (pCI-neo/Ctrl) for studies. These findings provide compelling evidence that the mTORC1/rpS6 signal pathway exerted its effects to promote Sertoli cell BTB remodeling. This was mediated through changes in the organization of actin- and microtubule-based cytoskeletons, involving changes in the distribution and/or spatial expression of actin- and microtubule-regulatory proteins.
Actins/metabolism* ; Animals ; Blood-Testis Barrier/metabolism* ; Cells, Cultured ; Male ; Mechanistic Target of Rapamycin Complex 1/metabolism* ; Permeability ; Rats ; Ribosomal Protein S6/metabolism* ; Seminiferous Epithelium/metabolism* ; Sertoli Cells/metabolism* ; Signal Transduction/physiology*

OBJECTIVETo investigate the correlation of exogenous estrogens with the expression of FasL in Sertoli cells and the blood-testis barrier during the differentiation and maturation period of Sertoli cells, and to discuss the related factors that influence the blood-testis barrier of pubertal rats.

METHODSSuper-physiological doses of exogenous estrogenic compounds (diethylstilbestrol and estradiol) were administered to pubertal Sprague-Dawley rats in vitro and in vivo, the FasL expression in the Sertoli cells of the rats detected by immunohistochemistry and Western blot, and the changes in the blood-testis barrier observed with the electron microscope.

RESULTSAfter the exposure to exogenous estrogens, the FasL expression was markedly up-regulated in the immature Sertoli cells (P < 0.05) as well as in the Sertoli cell membrane and the blood-testis barrier of the epithelium. The tracer lanthanum passed through the blood-testis barrier and reached the whole layer of the epithelium at 18 days.

CONCLUSIONSuper-physiological dose of exogenous estrogens can change the expression and distribution of FasL in immature Sertoli cells and affect the structure of the blood-testis barrier.

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Blood-Testis Barrier ; drug effects ; metabolism ; Estrogens ; pharmacology ; Fas Ligand Protein ; biosynthesis ; Male ; Models, Animal ; Rats ; Rats, Sprague-Dawley ; Sertoli Cells ; cytology ; drug effects ; metabolism

Infertility is a world wide problem affecting up to 15% of married couples. Although it is well known that male factor is the important cause of the infertility in 40-50% of the cases, the appropriate drugs for treating this condition have not yet been established. With certain exceptions, the etiology of many cases of male infertility is unknown. For such cases, various drugs including both hormonal and non-hormonal agents are sometime prescribed, but there have been no entirely satisfactory results. The present investigation would assess the effectiveness of Korean Ginseng, herbal medicine in the treatment of male infertility during the period from September to December, 1988, at the Andrology Clinic of the Department of Urology, Seoul National University Hospital. This herbal medicine was selected because its ingredients accelerate the metabolism of lipids and synthesis of DNA, RNA and proteins. This medicine contains ingredients which build resistance against stress since many of male infertility are under stress and also is to control immunological disorders. Ginseng has steroid-like, anti-allergic and anti-inflammatory actions, and accommodates the blood-testis barrier, improve digestive functions and peripheral blood flow. Accordingly, Ginseng has been used as an agent restoring healthy conditions to maintain homeostasis or to keep physical and mental balance. Extensive chemical investigation on Ginseng has revealed that Ginseng contains characteristic dammarane-type triterpenoid saponins as the main principles. These saponins are called ginsenosides and represent the principal pharmacological actions of Ginseng. The ginsenosides react at the hypothalamus-pituitary-gonadal axis. The decreases of sexual drive and disorders of fecundity under challenge of stress are prevented by oral administration of ginsenosides. To assess the efficacy of treatment with Ginseng which is alleged to improve spermatogenesis in idiopathic infertile selected patients. Participants in this study are men with primary idiopathic oligospermia and asthenospermia. The inclusion criteria would be as follows : a) men aged 20-40 years, whose female partners are entirely normal. b) men having vaginal intercourse with one partner and without psycho-sexual problems. c) men willing to enter this clinical trial and relying only on the drug administered throughout the study. d) no history of serious chronic physical or psychological diseases. e) men whose female partners are not using any method of birth control. f) men with no history of drugs to treat sperm disorders within 3 months. A total of 12 patients with sperm counts of less than 20 x 10 6/ml (oligospermia group), and 5 patients with sperm motility of less than 20% (asthenospermia group) are selected as the study subjects. Parameters to be assessed are as follows : Before and after Ginseng administration, history taking, physical examination with testis size measurement, laboratory works including urinalysis, CBC, seminal fructose, semen analyses (pH, volume, density, motility, activity grade, morphology, fertility unit, and WBC), plasma hormonal assays (FSH, LH, testosterone, estradiol, and prolactin). Before starting the treatment, 2 semen samples are obtained preteded by 3 days of abstinence. For follow-up, patients will have a semen sample taken every month while in treatment. After the treatment, more than 2 semen analyses will be undertaken for the final evaluation. Treatment scheme is as follows : The composition of the Ginseng extract used in this clinical trial consisted of the standardized highly concentrated Ginseng extract G115, 100mg : concentrated standardized lectithin, 95mg , alpha tocopherol, 10mg ; and excipients q.s.ad. This Ginseng extract named Ginsana capsule produced by Pharmaton-Korea Co., Ltd. Four capsules of Ginsana are given twice a day by mouth before meal for more than 90 days to be justified on the basis of general assumption that spermatogenesis cycle lasts approximately 74 days. The results of the clinical investigation are considered to be effective, if more than double of improvemant being noted on the count or more than 30% ot improvement being noted on motility beyond the pre-treatment levles. Clinical characteristics of a total of 17 patients are listed in the table 1. The outcomes of this trial are presented as follows : (tables 2 and 3). Coital frequency increased from 2.6/week before Ginsana exposure to 3.1/week after the treatment. General health such as stamina, body weight and spirits improved in 10 patients of the 17 after Ginsang treatment. Regarding hormonal partmeters (table 2), Plasma FSH and LH were not changed much before and after Ginsana administration. Patients with low FSH and LH levels before the treatment and patients with high range of prolactin levels before the treatment have a tendency to improve more in semen parameters after the treatment. Hyperestrogenemia was decreased and plasma testosterone levels increased after Ginsana treatment. Subsequently, T/E2 ratio resulted in normal to help spermatogenesis. Regarding the semen parameters (table 3), sperm counts increased in 58% of the patients in oligosperrnia group after oligospermia group. Sperm motility improved in 33% of the patients rn oligospermia group after the treatment. Mean motility increased from 34% to 45% after the treatment in oligospermia group. Activity grade and fertility unit were also improved in oligospermia group after the treatment Other parameters such as volume, normal morphology, pH and seminal fructose were not changed significantly before and after Ginstna treatment. Only one case showed an improvement in sperm counts and motility of a total of 5 patients with asthenospermia. Pregnancy resulted in 2 patients of improved cases and 1 patient of not improved cases in oligopsermia group after Ginsana administration. So that, pregnancy rate was 25 % of the oligospermia group. The study results of some imvestigators are summarized in the table 4. From these results, Ginsana appears mainly act on testis directly, restore the steroidogenesis, resulting in the stimulation of spermatogenesis. in conclusion, the authors clinical experience confirmed that Ginsana, a traditional Chinese medicine, appears to be of value particularly in the trettment of idiopethic oligospermia without any noticeabel adverse side effects.
Administration, Oral ; alpha-Tocopherol ; Andrology ; Axis, Cervical Vertebra ; Blood-Testis Barrier ; Body Weight ; Capsules ; Coitus ; Contraception ; DNA ; Estradiol ; Excipients ; Family Characteristics ; Female ; Fertility ; Fibrinogen ; Follow-Up Studies ; Fructose ; Ginsenosides ; Herbal Medicine ; Homeostasis ; Humans ; Hydrogen-Ion Concentration ; Infertility ; Infertility, Male ; Male ; Meals ; Medicine, Chinese Traditional ; Metabolism ; Mouth ; Oligospermia* ; Panax* ; Physical Examination ; Plasma ; Pregnancy ; Pregnancy Rate ; Prolactin ; RNA ; Saponins ; Semen ; Semen Analysis ; Seoul ; Sperm Count ; Sperm Motility ; Spermatogenesis ; Spermatozoa ; Testis ; Testosterone ; Tocopherols ; Urinalysis ; Urology