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

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*

Previous concepts of immune-privileged sites obscured the role of peripheral immune cells in neurological disorders and excluded the consideration of the potential benefits of immunotherapy. Recently, however, numerous studies have demonstrated that the blood-brain barrier in the central nervous system is an educational barrier rather than an absolute barrier to peripheral immune cells. Emerging knowledge of immune-privileged sites suggests that peripheral immune cells can infiltrate these sites via educative gates and that crosstalk can occur between infiltrating immune cells and the central nervous system parenchyma. This concept can be expanded to the testis, which has long been considered an immune-privileged site, and to neurogenic bladder dysfunction. Thus, we propose that the relationship between peripheral immune cells, the brain, and the urologic system should be considered as an additional possible mechanism in urologic diseases, and that immunotherapy might be an alternative therapeutic strategy in treating neurogenic bladder dysfunction.
Blood-Brain Barrier ; Brain ; Central Nervous System ; Immune System* ; Immunotherapy ; Nervous System Diseases* ; Testis ; Urinary Bladder, Neurogenic ; Urologic Diseases ; Urology*

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

Spermatogenesis is a complex regulatory process depending on a variety of hormones (such as FSH, LH, T, and 17beta estradiol), cytokines, and genes. Research on gene regulation in spermatogenesis has become a hot spot and revealed some spermato-genesis-related genes, such as AYZ, DAZ, YRRM, NOSTRIN, and so on. Reports are rarely seen on the role of CR16 in male reproduction, and its action mechanism in spermatogenesis is not yet clear. This article updates the role of CR16 in spermatogenesis in the male reproductive system from the perspective of Sertoli cells forming a blood-testis barrier.
Animals ; Blood-Testis Barrier ; Humans ; Male ; Microfilament Proteins ; Sertoli Cells ; Spermatogenesis ; Testis ; cytology

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

PURPOSE: Our study aimed to determine whether the severity of damage to the contralateral testis by ipsilateral testicular torsion/detorsion in pubertal rats, which have an incomplete blood-testis barrier, is different from that in adult rats. MATERIALS AND METHODS: We divided pubertal (6 weeks, n=17) and adult (10 weeks, n=17) Sprague-Dawley (SD) rats into group S (sham; n=5), group O (orchiectomy; n=6), and group D (detorsion; n=6). After 4 hours' torsion of the ipsilateral testis, we applied orchiectomy (group O) and detorsion (group D) depending on the group and compared the histopathologic changes and germ cell apoptosis of the contralateral testis at the age of 13 weeks. RESULTS: In each age group, increased interstitial area, edema, and germ cell sloughing were observed in group D. The mean seminiferous tubule diameter decreased more in group D than in group S or O in each age group (p<0.05). The mean germ cell layer thickness and number of spermatids per tubule decreased more in group D than in group S or O in each age group; additionally, in group D, values decreased more in pubertal rats than in adult ones (p<0.05, respectively). The mean numbers of terminal deoxyuridine nick-end labeling (TUNEL)-positive cells were less than 1.0 in groups S and O, which was smaller than in group D (p<0.05); additionally, in group D, this value tended to be higher in pubertal rats than in adult ones (p=0.057). CONCLUSIONS: SD rats with a detorsioned testis had more severe damage to the contralateral testis than did those undergoing orchiectomy of the torsioned testis. Also, when comparing the severity of damage to the contralateral testis after ipsilateral torsion/detorsion between pubertal and adult rats, rats at a pubertal age, when most testicular torsions occur in clinical situations, had more severe damage than did those at an adult age.
Adult ; Age Factors ; Animals ; Apoptosis ; Blood-Testis Barrier ; Deoxyuridine ; Edema ; Germ Cells ; Humans ; Orchiectomy ; Rats ; Seminiferous Tubules ; Spermatic Cord Torsion ; Spermatids ; Testis

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 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

OBJECTIVETo determine the effect of high power microwave (HPM) radiation on the structure and function of blood-testis barrier (BTB) in rats.

METHODSOne hundred and sixty-six male Wistar rats were treated by heart perfusion of lanthanum-glutaraldehyde solution and tail vein injection of evans blue (EB) at 6 h, 1, 3, 7 and 14 d after exposed to 0, 10, 30 and 100 mW/cm2 HPM radiation for 5 minutes, the structural change of BTB and distribution of lanthanum or EB observed through the light microscope, electron microscope and laser scanning confocal microscopy (LSCM).

RESULTSTesticular interstitial edema, vascular congestion or hyperemia with accumulation of plasma proteins and red blood cells in the inner compartment of seminiferous tubules were observed after exposure to HPM. The above-mentioned pathological changes were aggravated at 1-7 d and relieved at 14 d after radiation, obviously more severe in the 30 and 100 mW/cm2 exposure groups than in the 10 mW/cm2. Both lanthanum precipitation and EB were deposited in the inner compartment.

CONCLUSIONHPM radiation may damage the structure and increase the permeability of BTB.

Animals ; Blood-Testis Barrier ; pathology ; physiopathology ; radiation effects ; Male ; Microwaves ; adverse effects ; Rats ; Rats, Wistar