OBJECTIVETo explore the effect of vinyl chloride on reproductive and endocrine system of male rats.

METHODSMale SD rats were administered with vinyl chloride at dose of (0, 10, 100, 1000 mg/kg) for 14 and 28 days, respectively. The levels of testosterone (T), inhibin B, luteinizing hormone (LH), follicle stimulating hormone (FSH) and estradiol (E2) were measured in serum and testis homogenates. Histopathological examinations were performed for testis with electron microscopy.

RESULTSCompared with the control group after 14-day exposure, T and E2 serum levels of 1000, 100, 10 mg/kg groups decreased, InhB and LH levels of three dose groups increased. LH serum levels of 100 mg/kg increased significantly statistically compared with control group (P < 0.05). After 28-day exposure, T serum levels of 100, 1000 mg/kg groups were (10.90 +/- 1.56), (8.52 +/- 2.85) ng/ml respectively (P < 0.05), InhB serum levels of 100, 1000 mg/kg groups were (31.40 +/- 6.21), (28.39 +/- 5.67) pg/ml respectively. Both of T and InhB serum levels of 100, 1000 mg/kg groups decreased significantly (P < 0.05). Serum FSH levels of 10, 100, 1000 mg/kg groups decreased significantly compared with control group (P < 0.05). Compared with groups of 14-day exposure, serum InhB and LH levels of 10, 100, 1000 mg/kg groups decreased significantly statistically after 28 days. T and InhB testis levels of 100, 1000 mg/kg groups were 8.05 +/- 2.19),(6.75 +/- 1.94) ng/mg pro and (39.32 +/- 5.55), (35.53 +/- 8.71) pg/mg pro respectively, which decreased significantly compared with control group (P < 0.05). Leydig cell and Sertoli cell were damaged according to histopathological examinations.

CONCLUSIONVinyl chloride has adverse effects on reproductive and endocrine system of male rats and may change their serum and testis homogenate levels of hormones.

Animals ; Follicle Stimulating Hormone ; blood ; Leydig Cells ; ultrastructure ; Male ; Rats ; Sertoli Cells ; ultrastructure ; Testis ; drug effects ; metabolism ; Testosterone ; blood ; Vinyl Chloride ; toxicity

OBJECTIVETo investigate the effect of baicalein on the gap junction intercellular communication (GJIC) in the TM4 Sertoli cells of the mouse testis and its related mechanism.

METHODSWe measured the cytotoxicity of different concentrations of baicalein on the TM4 Sertoli cells in the mouse testis by MTT, detected the fluorescence transfer of the TM4 Sertoli cells by parachute assay, and determined the expression of the protein connexin 43 ( Cx43) in the baicalein-treated cells by Western blot and immunofluorescence assay.

RESULTSBaicalein produced no obvious cytotoxicity on the TM4 Sertoli cells at the concentration below 60 µmol/L but significantly increased their GJIC at 0-20 µmol/L (P < 0.01). Western blot and immunofluorescence assay showed that 0-20 µmol/L baicalein remarkably elevated the expression of Cx43 in the TM4 cells (P < 0.01) and on the membrane of the TM4 cells.

CONCLUSIONBaicalein at the concentration of 0-20 µmol/L can significantly enhance GJIC in mouse TM4 Sertoli cells by increasing the expression of the Cx43 protein.

Animals ; Cell Communication ; drug effects ; Connexin 43 ; metabolism ; Flavanones ; administration & dosage ; pharmacology ; Gap Junctions ; drug effects ; Male ; Mice ; Sertoli Cells ; drug effects ; metabolism ; ultrastructure

Actin can be found in all kinds of eukaryotic cells, maintaining their shapes and motilities, while its dynamics in sperm cells is understood less than their nonmuscle somatic cell counterparts. Spermatogenesis is a complicated process, resulting in the production of mature sperm from primordial germ cell. Significant structural and biochemical changes take place in the seminiferous epithelium of the adult testis during spermatogenesis. It was proved that all mammalian sperm contain actin, and that F-actin may play an important role during spermatogenesis, especially in nuclear shaping. Recently a new model for sperm head elongation based on the acrosome-acroplaxome-manchette complex has been proposed. In Drosophila, F-actin assembly is supposed to be very crucial during individualization. In this mini-review, we provide an overview of the structure, function, and regulation characteristics of actin cytoskeleton, and a summary of the current status of research of actin-based structure and movement is also provided, with emphasis on the role of actins in sperm head shaping during spermiogenesis and the cell junction dynamics in the testis. Research of the Sertoli ectoplasmic specialization is in the spotlight, which is a testis-specific actin-based junction very important for the movement of germ cells across the epithelium. Study of the molecular architecture and the regulating mechanism of the Sertoli ectoplasmic specialization has become an intriguing field. All this may lead to a new strategy for male infertility and, at the same time, a novel idea may result in devising much safer contraception with high efficiency. It is hoped that the advances listed in this review would give developmental and morphological researchers a favorable investigating outline and could help to enlarge the view of new strategies and models for actin dynamics during spermatogenesis.
Acrosome ; physiology ; Actins ; chemistry ; physiology ; Animals ; In Vitro Techniques ; Intercellular Junctions ; physiology ; Male ; Sertoli Cells ; physiology ; ultrastructure ; Sperm Motility ; physiology ; Spermatogenesis ; physiology ; Testis ; cytology ; physiology

OBJECTIVETo establish the testis transplantation model in rats and to study the mechanism of graft injury.

METHODSThe testis orthotopic transplantation model was established using three-cuff method. The animals were divided into 6 groups. Serum levels of testosterone (T), luteining hormone (LH) and follicle-stimulating hormone (FSH) were determined by radioimmunoassay (RIA). Morphology and ultrastructure were examined by light and electron microscopy. Expression of Glial cell line-derived neurotrophic factor (GDNF) mRNA was studied by reverse-transcription polymerase chain reaction (RT-PCR) technique.

RESULTOn the 7th day postoperatively, the allotransplanted testes showed perivascular massive infiltration of lymphocytes and polymorphonuclear neutrophil (PMN) and reduced number of the sertoli cells under light microscopy. It also showed the broken blood-testis barrier, the atrophy of the sertoli cells and spermatogenic cells arranged in disorder under electron microscopy. The decline of serum T level and the increase of serum LH and FSH levels were similar to those found in bilateral castrates. The levels of GDNFmRNA expression were lower than those in normal controls. On 14th day postoperatively, the spermatogenesis of allotransplanted testes was still not recovered and the expression of GDNFmRNA declined further.

CONCLUSIONThe atrophy and reduced number of the sertoli cells and the breakage of the close connection probably are the main causes of dysfunction of spermatogenesis. The decline of GDNFmRNA expression is in accordance with the dysfunction of the sertoli cells and the spermatogenesis.

Animals ; Follicle Stimulating Hormone ; blood ; Glial Cell Line-Derived Neurotrophic Factor Receptors ; biosynthesis ; genetics ; Luteinizing Hormone ; blood ; Male ; Models, Animal ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Inbred Lew ; Rats, Wistar ; Sertoli Cells ; ultrastructure ; Spermatogenesis ; physiology ; Testis ; transplantation ; ultrastructure ; Testosterone ; blood

In this study, the effects of mono(2-ethylhexyl) phthalate (MEHP), one of metabolites of di(2-ethylhexyl) phthalate, on immature Shiba goat testes in vitro were examined. The testes of 2-month-old Shiba goats were cut into smaller pieces, and seeded in medium. At 1, 3, 6 and 9 hr after administration of MEHP at various concentrations (0, 100 nmol ml-1, 1 nmol ml-1, and 1 x 10-3 nmol ml-1, respectively), the specimens were obtained for light and transmission electron microscopic observations. As a result, at 1 hr after exposure to MEHP, the vacuolization and nuclear membrane rupture appeared in Sertoli cells. Such alterations tended to gradually increase in number in timeand dose-dependent manners. Moreover, by MEHP treatment, apoptotic spermatogenic cells (characterized with chromatin condensation, cytoplasm shrinkage without membrane rupture, still functioning cell organelles, and packed cell contents in membrane-bounded bodies), apoptotic Sertoli cells (characterized with nuclear membrane lysis, nuclear condensation), necrotic spermatogenic cells (characterized with swollen and ruptured mitochondria, plasma membrane lysis, spilt cell contents, and chromatin clumps), and necrotic Sertoli cells (characterized with marginated chromatins along the nuclear membrane, ruptured vesicles within the MNB, some swollen and ruptured cell organelles, e.g. mitochondria) could be identified. Conclusively, ultrastructurally the treatment with MEHP at low concentration tends to lead spermatogenic and Sertoli cells to apoptosis, whereas that at high concentration tends to lead spermatogenic and Sertoli cells to necrosis. Thus, the testicular tissue culture is advantageous for screening testicular toxicity of chemicals.
Animals ; Apoptosis/drug effects/physiology ; Diethylhexyl Phthalate/*analogs&derivatives/*toxicity ; Goat Diseases/*chemically induced/pathology ; Goats ; Male ; Microscopy, Electron, Transmission/veterinary ; Necrosis ; Sertoli Cells/ultrastructure ; Spermatozoa/drug effects/pathology/ultrastructure ; Testicular Diseases/*chemically induced/pathology ; Testis/*drug effects/metabolism/pathology/ultrastructure ; Vacuoles/physiology/ultrastructure

AIMTo evaluate the key lesions in spermatogenesis suppressed partially by testosterone undecanoate (TU) treatment.

METHODSAdult male SD rats were treated with vehicle or TU (19 mg/kg) injection (i.m.) every 15 days for 130 days. The numbers of all types of cells (nuclei) in the seminiferous tubules and the interstitial tissue were estimated using a contemporary stereological tool, the optical disector.

RESULTSIn response to TU treatment, the numbers of non-type B spermatogonia, type B spermatogonia and late elongated spermatids per testis were reduced to 51 %, 66 % and 14 % of the controls, respectively. The conversion ratios from type B spermatogonia to early spermatocytes and pachytene spermatocytes were not significantly affected and the ratios to the later germ cell types fell to 51 % - 65 % of the controls. Less than 1.0 % of immature round spermatids were seen sloughing into the tubule lumen, 4.0 % of elongated spermatids retained in the seminiferous epithelium, and about half of the elongated spermatid nuclei appreciably malformed. Leydig cells were atrophied but their number and the peritubular myoid cell number per testis were unchanged.

CONCLUSIONDouble inhibition of spermatogenesis (i.e. inhibition at spermiation and spermatogonial conversion to type B spermatogonia), a scenario seen in the monkey and human following gonadotrophin withdrawal, was not sufficiently effective for a complete spermatogenic suppression in the rat after TU treatment, probably due to ineffective inhibition of the Leydig cell population and therefore the intra-testicular testosterone levels.

Animals ; Cell Nucleus ; drug effects ; ultrastructure ; Depression, Chemical ; Leydig Cells ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Sertoli Cells ; drug effects ; Sperm Count ; Spermatids ; drug effects ; Spermatogenesis ; drug effects ; Testosterone ; analogs & derivatives ; blood ; pharmacology

OBJECTIVETo simplify the method for separation and cultivation of rat testicular Sertoli cells with high viability, quantity and expression efficiency.

METHODSTesticular Sertoli cells from 2 to 3-week-old male Wistar rats were prepared by digestion with collagenase, trypsin and DNase and cultured together with active lymphocytes to observe their killing effect against lymphocytes. After cell culture for 72 h, the Sertoli cells were morphologically observed by different means and identified with transmission electron microscope. Fas ligand and follicle-stimulating hormone receptor (FSHR) were examined immunohistochemically to identify testicular Sertoli cells. SABC method was used for labeling the Fas ligand on the testicular Sertoli cells.

RESULTSThe viability of the isolated and cultured Sertoli cells was more than 90%, and in in vitro culture, Sertoli cells, which expressed the Fas ligand, could kill the active lymphocytes.

CONCLUSIONThis method improves the efficiency in acquisition of rat testicular Sertoli cells expressing Fas ligand, which are believed to be a potential donor for co-transplantation with parathyroid cells to offer immune privilege.

Animals ; Cell Communication ; immunology ; Cell Separation ; methods ; Cell Survival ; immunology ; Cells, Cultured ; Fas Ligand Protein ; metabolism ; Immunohistochemistry ; Lymphocytes ; cytology ; immunology ; Male ; Microscopy, Electron, Transmission ; Rats ; Rats, Wistar ; Receptors, FSH ; metabolism ; Sertoli Cells ; cytology ; metabolism ; ultrastructure ; Testis ; cytology

OBJECTIVETo explore the synergistic protective effect of co-transplanted testicular cells expressing FasL and CsA on survival of islet allografts.

METHODSThe allogeneic islets and testicular cells were co-transplanted into the renal subcapsular space of the diabetic recipients with or without CsA after operation. Allografts survival period and the testicular cells or islets function were analyzed.

RESULTSThe mean survival period of control group was 4.6 +/- 1.1 days. When CsA was administered after transplantation, the mean survival period of islet allografts, (21.8 +/- 4.7) days, was significantly longer than that of control group (P < 0.01). When islets were co-transplanted together with 1 x 10(7) testicular cells (group A), a significant prolongation of graft survival was found (more than 57.5 +/- 4.0 days; P < 0.01 vs. control). But if 1 x 10(7) testicular cells expressing FasL were cultured with FasL-mAb for 30 minutes before co-transplantation (group B), the mean survival period of islet allografts (5.8 +/- 2.6 days), was similar to that in control group, but significantly shorter than that in group A (P < 0.01). When islets and 1 x 10(5) testicular cells were co-transplanted separately into the bilateral renal subcapsular space with CsA (group C), the survival of islet allografts was significantly prolonged in comparison with control group (more than 55.0 +/- 6.5 days; P < 0.01 vs. control), and similar to islets co-transplanted together with 1 x 10(7) testicular cells (group A). When islets were co-transplanted separately with 1 x 10(6) testicular cells without CsA (group D), the mean survival period (11.5 +/- 3.1 days) was shorter than that in group C, but prolonged in comparison to control group (P < 0.05).

CONCLUSIONThe co-transplanted testicular cells expressing FasL with administering CsA post-transplantation can jointly inhibit immune rejection of islet allografts by different mechanism and play a systemic and synergistic protective role to islet allografts.

Animals ; Cyclosporine ; therapeutic use ; Fas Ligand Protein ; Graft Survival ; Immunohistochemistry ; Insulin ; blood ; Islets of Langerhans ; pathology ; ultrastructure ; Islets of Langerhans Transplantation ; Male ; Membrane Glycoproteins ; analysis ; genetics ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Sertoli Cells ; metabolism ; transplantation ; Transplantation, Homologous

OBJECTIVETo study the ultrastructural changes of the rat convoluted seminiferous tubule after alcohol consumption.

METHODSForty-eight Wistar mature male rats were divided into two groups randomly: control group (A) and experimental one (B). 6 ml/(kg x d) of 50 degrees alcohol was perfused through the gastric tube for 39 days in Group B; and 6 ml/(kg x d) of normal saline was supplemented in Group A. The ultrastructure of the rat convoluted seminiferous tubule was observed by transmission electron microscope at day 14, 27 and 40.

RESULTSIn Group A, the pykno-basement membrane was unstriated and uniform, Sertoli cells showed cytoplasmic profusion, with big nucleus, well-distributed nucleoplasm, distinct nucleolus, more mitochondria and plain hierarchical tight-junction. And the ultrastructure of the rat convoluted seminiferous tubule in Group B began to change at the end of the first spermatogenic cycle (D 14) and changed more and more evidently with the ethanol administration, mainly as follows: (1) more lysosomes and vacuolisation found in Sertoli cells, and organelles decreased and blurry; (2) more and bigger vacuoles among the spermatogonia, Sertoli cells and basement membrane; (3) obvious apoptosis of spermatogonia and apoptotic bodies aggregated near the membrane; (4) more cytoplasm and vacuolisation in the sperm of the convoluted seminiferous tubule, and disarranged, deleted or clustered mitochondria in the sperm tail; (5) blurry and rigid tight-junction; (6) thickened, wrinkled or broken basement membrane and under-basement

CONCLUSIONAlcohol can cause ultrastructural changes of the basement membrane, tight-junction and Sertoli cells of the membrane. rat convoluted seminiferous tubule and apoptosis of spermatogonia.

Animals ; Apoptosis ; drug effects ; Basement Membrane ; drug effects ; pathology ; Ethanol ; toxicity ; Male ; Microscopy, Electron, Transmission ; Random Allocation ; Rats ; Rats, Wistar ; Seminiferous Tubules ; drug effects ; ultrastructure ; Sertoli Cells ; drug effects ; pathology

ObjectiveTo investigate the influence of cellphone electromagnetic radiation (CER) on the testicular ultrastructure and the apoptosis of spermatogenic cells in male rats.atability, feasibility, applicability, and controllability in the construction of experimental animal models, we compared the major anatomic features of the penis of 20 adult beagle dogs with those of 10 adult men. Using microsurgical techniques, we performed cross-transplantation of the penis in the 20 (10 pairs) beagle dogs and observed the survival rate of the transplanted penises by FK506+MMF+MP immune induction. We compared the relevant indexes with those of the 10 cases of microsurgical replantation of the amputated penis.

METHODSThirty adult male SD rats were equally randomized into a 2 h CER, a 4 h CER, and a normal control group, the former two groups exposed to 30 days of 900 MHz CER for 2 and 4 hours a day, respectively, while the latter left untreated. Then the changes in the ultrastructure of the testis tissue were observed under the transmission electron microscope and the apoptosis of the spermatogenic cells was determined by TUNEL.

RESULTSCompared with the normal controls, the rats of the 2 h CER group showed swollen basement membrane of seminiferous tubules, separated tight junction of Sertoli cells, increased cell intervals, apparent vacuoles and medullization in some mitochondria, and increased apoptosis of spermatogenic cells, mainly the apoptosis of primary spermatocytes (P<0.05 ). In comparison with the 2 h CER group, the animals of the 4 h CER group exhibited swollen basement membrane of seminiferous tubules, more separated tight junction of Sertoli cells, wider cell intervals, incomplete membrane of spermatogonial cells, fragments of cytoplasm, nuclear pyknosis and notch, slight dilation of perinuclear space, abnormalities of intracellular mitochondria with vacuoles, fuzzy structure, and fusion or disappearance of some cristae, and increased damage of mitochondria and apoptosis of spermatogenic cells, including the apoptosis of spermatogonial cells, primary spermatocytes, and secondary spermatocytes (P<0.05 ).

CONCLUSIONSCER can damage the testicular ultrastructure and increase the apoptosis of spermatogenic cells of the male rat in a time-dependent manner, and the apoptosis of spermatogenic cells may be associated with the damage to mitochondria.

Animals ; Apoptosis ; Cell Phone ; Electromagnetic Radiation ; Male ; Mitochondria ; radiation effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Seminiferous Tubules ; radiation effects ; Sertoli Cells ; radiation effects ; Spermatocytes ; radiation effects ; Spermatogonia ; radiation effects ; Testis ; radiation effects ; ultrastructure