Plasminogen activator(PA) and plasminogen activator inhibitor(PAI) are involved in many physiological or pathological events. The Sertoli cells, the important elements within the seminiferous epithelium, are thought to play a key role in spermatogenesis. The Sertoli cells secrete PA and PAI. The levels of them are modulated by hormonal and cell-mediated influences. They play a fundamental role in the maintenance of spermatogenesis, sperm motility and fertilization.
Humans ; Male ; Plasminogen Activators ; metabolism ; physiology ; Plasminogen Inactivators ; metabolism ; physiology ; Sertoli Cells ; metabolism ; physiology ; Testis ; cytology

OBJECTIVETo explore the action mechanisms of temperature in male infertility or subfertility by observing the effects of different temperatures on the proliferation of and occludin (OCLN) expression in rat Sertoli cells in vitro.

METHODSWe isolated Sertoli cells from the testis of male Wistar rats, and performed oil red O staining and immunohistochemistry to identify their FasL. We cultured the Sertoli cells at 34 degrees C (control group) and at 35, 36, 37, 38 and 39 degrees C (experimental groups) for 4 days. Then we measured their proliferation by CCK-8 assay, observed their morphology and structure by hematoxylin-eosin staining, and determined their OCLN expression level by Western blotting and immunofluorescence.

RESULTSThe purity of the isolated Sertoli cells was (96.20 +/- 1.95)%. CCK-8 assay indicated that the proliferation of the Sertoli cells was increased between 34 and 36 degrees C, and decreased at 36-39 degrees C. The pyknotic nuclei and fragmentation of the Sertoli cells were more obvious at > 36 degrees C. Western blot and immunofluorescence showed the highest level of OCLN expression at 36 degrees C, which, however, decreased while the temperature rose above 36 degrees C (P < 0. 01).

CONCLUSIONHigh temperature (> 36 degrees C) inhibited the proliferation of rat Sertoli cells in vitro, and decreased the expression of OCLN, which suggests that a higher temperature above 36 degrees C may reduce male fertility by affecting the proliferation of Sertoli cells and integrity of the tight junction among Sertoli cells or Sertoli cells and other cells.

Animals ; Cell Proliferation ; Male ; Occludin ; metabolism ; Rats ; Rats, Wistar ; Sertoli Cells ; cytology ; metabolism ; Temperature ; Testis ; cytology ; metabolism

This article introduces the structure and function of the Sertoli cell cytoskeleton of the testis and the research progress in this aspect, focusing on the description of the function of vimentin, with some illustrations on the impact of physical and chemical factors on cytoskeleton, especially the structural changes of vimentin cell microfilament under simulated microgravity and space true microgravity. It for the first time proposes that the Sertoli cell cytoskeleton can be detected in semen, with a view to involving more researchers in further studies in this field.
Actin Cytoskeleton ; metabolism ; physiology ; Animals ; Apoptosis ; physiology ; Cytoskeleton ; metabolism ; physiology ; Humans ; Male ; Mice ; Rats ; Semen ; cytology ; metabolism ; Sertoli Cells ; cytology ; metabolism ; Testis ; cytology ; metabolism ; Vimentin ; metabolism

OBJECTIVETo explore the characteristics and distribution of GATA-4 in the testis of male mice.

METHODSParaffin sections were obtained from the testes of 24 male B6SJLF1/J mice, aged 0 day (n = 6), 2 weeks (n = 6), 4 weeks (n = 6) and 6 weeks (n = 6), and the expressions of GATA-4 in the testis were observed by the immunohistochemical ABC method and DAB visualization at different times.

RESULTSPositive expressions of GATA4 were found in the Sertoli cells and Leydig cells of all the mice, but significantly higher in the 4- and 6-week-old than in the 0-day and 2-week-old groups (P < 0.01). And they were also observed in the germ cells of the 4- and 6-week-old mice, significantly higher in the latter than in the former (P < 0.01).

CONCLUSIONGATA-4 exists in the testis of male mice, which has provided a morphological base for sex determination and differentiation and hormone regulation in the testis.

Animals ; Cell Differentiation ; GATA4 Transcription Factor ; metabolism ; Germ Cells ; metabolism ; Leydig Cells ; metabolism ; Male ; Mice ; Sertoli Cells ; metabolism ; Testis ; cytology ; metabolism

There has been no study aimed at directly determiningof the periods of Sertoli cell proliferation in birds evendomestic fowl. The aims of this study were to observe thecessation of post-hatching mitotic proliferation of Sertolicells in domestic fowl, and to determine the volumedensity of Sertoli and germ cells during this period. Atotal of 50 Leghorn chicks were used in this study. Thetestes sections of the animals were immunostained withBrdU to observe the proliferation of cells from one to 10weeks of age. The volume density of the Sertoli and germcells were determined using the standard point countingmethod. The volume density of the germ cell nuclei wasinitially less than that of the Sertoli cells but the volumedensity converged by week 6, and remained relativelyconstant until the commencement of meiosis. Clearlabeling of Sertoli and germ cells was observed from week1 to week 7. The only those cells still labeled after 8 weekswere germ cells, indicating that Sertoli cell proliferationhad ceased. Therefore, it is recommended that anyresearch into the testes of domestic fowl should considerthe cessation of Sertoli cell proliferation by approximately8 weeks.
Animals ; Bromodeoxyuridine/metabolism ; Cell Differentiation/physiology ; Cell Growth Processes/physiology ; Chickens/*physiology ; Histocytochemistry/veterinary ; Male ; Mitosis/physiology ; Sertoli Cells/*cytology/metabolism ; Spermatocytes/cytology ; Testis/*cytology/metabolism

The expression of osteopontin (OPN) in boar testis was studied. Western blot analysis detected 66- and 32-kDa OPN immunopositive bands in the testes of adult boars. In postnatal piglets, the 66-kDa OPN band was detected in the testes, but not the 32-kDa band. In the newborn testis, OPN immunostaining was seen in gonocytes and in some supporting cells in the seminiferous tubules, as well as in interstitial Leydig cells. In the adult boar testis, OPN immunoreactivity was detected in seminiferous tubules with varying intensities. Intense OPN immunostaining was seen in the residual bodies and acrosomes in the spermatids while, occasionally, OPN immunostaining was seen in spermatogonia and various stage of spermatocytes but in few Sertoli cells in the seminiferous tubules. In addition, Leydig cells in adult boars were weakly immunostained with OPN. These findings suggest that OPN is detected in the majority of germ cells and is involved in spermatogenesis in boar testis.
Animals ; Animals, Newborn ; Blotting, Western/veterinary ; Immunohistochemistry/veterinary ; Leydig Cells/metabolism ; Male ; Osteopontin/*biosynthesis ; Sertoli Cells/metabolism ; Spermatogenesis/physiology ; Spermatozoa/metabolism ; Swine/*metabolism ; Testis/cytology/*metabolism

OBJECTIVETo explore the mechanism of hyperthermia inducing infertility by observing the expression of glial cell line-derived neurotrophic factor (GDNF) in rat Sertoli cells cultured in vitro at different temperatures.

METHODSUsing combination enzyme digestion and selective adhesion, we isolated Sertoli cells from male Wistar rats and cultured them in vitro at different temperatures, followed by observation of the changes in their adhesion and morphology and identification by FasL immunohistochemical staining. We divided the Sertoli cells into a control group (35 degrees C) and four experimental groups (36 degrees C, 37 degrees C, 38 degrees C, and 39 degrees C), measured their proliferation by CCK-8, observed their morphology and structure by HE staining, and determined the expression of GDNF by RT-PCR, immunofluorescence and Western blot.

RESULTSSertoli cells were successfully isolated and in vitro-cultured, with a purity of (95.30 +/- 2.15)% (n = 10). The CCK-8 assay showed that the proliferation of the Sertoli cells was the highest at 36 degrees C, gradually decreasing with the temperature above 36 degrees C, and significantly inhibited at 39 degrees C (P < 0.01). Immunofluorescence revealed the expression of GDNF in the cytoplasm, with the highest fluorescence intensity at 36 degrees C. RT-PCR and Western blot exhibited a decreasing trend of the GDNF expression with the increasing temperature above 36 degrees C. There were statistically significant differences in the expression of GDNF between the control group and the four experimental groups (P < 0.01).

CONCLUSIONThe proliferation and GDNF expression of in vitro-cultured Sertoli cells differ significantly at different temperatures. At > 36 degrees C, the higher the temperature is, the lower the Sertoli cell proliferation and GDNF expression are. Our findings suggest that high temperature above 36 degrees C suppresses the function of Sertoli cells and may also damage spermatogenesis.

Animals ; Cells, Cultured ; Glial Cell Line-Derived Neurotrophic Factor ; metabolism ; Male ; Rats ; Rats, Wistar ; Sertoli Cells ; cytology ; metabolism ; Temperature ; Testis ; cytology

Sertoli cell (SC) is intrinsic to the testis and provides an appropriate growth environment for the germ cells. It was separated from rat's testis and identified by hematoxylin and eosin staining(HE) and immunocytochemical reaction, then cultivated in vitro. Culture conditions such as pH, osmotic pressure and metabolic parameters that include consumption rates of glucose, glutamine, amino acids and formation rates of lactic acid, ammonium ion were investigated. It was showed that adhesion process of SCs was accomplished within 2-4 hours after inoculation. It was also observed that the SCs entered into the decline phase when the concentration of ammonium ion and lactic acid were above 2.3 mmol/L and 14 mmol/L, respectively, which caused osmotic pressure above 326 mosm/kg and pH below 6.8 in the medium. As the changes of amino acids during culture were concerned, Glu and Ala accumulated rapidly, while Val, Leu, Ile reduced slightly and at the same time Ser, Arg, and Gly were stable. The restrict factors for SCs grown in static culture might be high osmotic pressure and low pH, which were generated when glutamine and glucose were metabolized into lactic acid. The findings could be fundamental in the process optimization of large scale Sertoli cells in vitro culture.
Amino Acids ; metabolism ; Animals ; Animals, Newborn ; Cell Culture Techniques ; Cells, Cultured ; Culture Media ; Male ; Rats ; Sertoli Cells ; cytology ; metabolism ; Testis ; cytology

OBJECTIVETo investigate the expressions of cadherin molecules CDH18 and PCDH17 in normal and azoospermic human testes and their significance.

METHODSWe studied the routine pathological slices of normal and non-obstructive azoospermic human testis tissues for changes in the tight junction of Sertoli-germ cells, and identified the differential gene expression profiles of the normal and azoospermic testis tissues using cDNA microarrays containing multiple cadherin molecules. The results were confirmed by Western blot.

RESULTSAbnormal tight junction of the Sertoli-germ cells was observed in 37.5% of the azoospermic testis samples, and obvious changes were seen in the expressions of some cadherin molecules, with down-regulation of CDH18 and PCDH17.

CONCLUSIONCadherin molecules such as CDH18 and PCDH17 may play a certain role in the development and progression of azoospermia, which might be related with the abnormal tight junction of the Sertoli-germ cells.

Adult ; Azoospermia ; metabolism ; Cadherins ; metabolism ; Cells, Cultured ; Down-Regulation ; Gene Expression Profiling ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; Sertoli Cells ; metabolism ; Testis ; cytology ; metabolism ; Tight Junctions ; Young Adult

AIMTo examine the possible effect of heat treatment on expression of heat shock proteins (Hsps) 105, 70, and 60 in primary monkey Sertoli cells and to evaluate the possible signal pathways.

METHODSWestern blot analysis, real-time polymerase chain reaction (PCR), and confocal immunohistochemistry were used to analyze mRNA and protein levels of the Hsps in response to 43 degrees treatment of Sertoli cells isolated from pubertal monkey testes.

RESULTSStaining with Hoechst 33342 indicated Sertoli cells did not undergo apoptosis after heat treatment. Hsp105 was expressed in cytoplasm of untreated Sertoli cells. Both Hsp105 mRNA and protein levels were increased approximately 20-fold compared to those of the untreated controls at 12 h after heat treatment. Untreated Sertoli cells did not express Hsp70, but heat stress induced its expression in the cell cytoplasm. The time-course of changes in Hsp70 was similar to that of Hsp105. In contrast to Hsp105 and Hsp70, the change in Hsp60 expression was much less obvious. The protein level between 12 h and 48 h after heat treatment was only approximately 1.5-fold that of the untreated control. Extracellular regulated kinase (ERK) 1/2 inhibitor (U0126) or phosphoinositide kinase-3 (PI3K) inhibitor (LY294002) could partially block the response of Hsp105 and Hsp70 induced by heat treatment.

CONCLUSIONThese results indicate that the heat-induced expression of the three types of Hsp in monkey Sertoli cells might be regulated by ERK and/or PI3K signal pathways, but the profile of their expression is different, suggesting that they might have different regulatory functions in Sertoli cells.

Animals ; Apoptosis ; Base Sequence ; Cells, Cultured ; Cold Temperature ; DNA Primers ; Heat-Shock Proteins ; genetics ; metabolism ; Immunohistochemistry ; Macaca mulatta ; Male ; RNA, Messenger ; genetics ; Sertoli Cells ; cytology ; metabolism