• Clinical and Experimental Reproductive Medicine
• /
• v.49 no.1
• /
• pp.9-15
• /
• 2022

The angiotensin-converting enzyme 2 receptor (ACE2) appears to be widely expressed in cells in the testes, predominantly in spermatogonia, Sertoli cells, and Leydig cells, and its co-expression with transmembrane protease serine 2 (TMPRSS2) is essential for the entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). For this reason, the male reproductive system could be considered a potential target for SARS-CoV-2, as well as a possible reservoir of infection. However, to date, there is very little evidence about the presence of SARS-CoV-2 in semen and testicular samples. The aim of this paper was to review the current evidence regarding the impact of SARS-CoV-2 on male fertility and sexual health, with a particular focus on reproductive hormones, the presence of the virus in seminal fluid and testis, and its impact on fertility parameters. We found very limited evidence reporting the presence of SARS-CoV-2 in semen and testicular samples, and the impact of SARS-CoV-2 on reproductive hormones and fertility parameters is unclear. The quality of the examined studies was poor due to the small sample size and several selection biases, precluding definitive conclusions. Hence, future well-designed prospective studies are needed to assess the real impact of SARS-CoV-2 on male reproductive function.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.9
• /
• pp.1347-1354
• /
• 2006

The aims of this study were to establish a simple and effective method for isolating male germline stem cells (GSCs), and to test the possibility of using these cells as a new approach for male infertility treatment. Testes obtained from neonatal and adult mice were manually decapsulated. GSCs were collected from seminiferous tubules by a two-step enzyme digestion method and plated on gelatin-coated dishes. Over 5-7 days of culture, GSCs obtained from neonates and adults gave rise to large multicellular colonies that were subsequently grown for 10 passages. During in vitro proliferation, oct-4 and two immunological markers (Integrin ${\beta}1,\;{\alpha}6$) for GSCs were highly expressed in the cell colonies. During another culture period of 6 weeks to differentiate to later stage germ cells, the expression of oct-4 mRNA decreased in GSCs and Sertoli cells encapsulated with calcium alginate, but the expression of c-kit and testis-specific histone protein 2B(TH2B) mRNA as well as the localization of c-kit protein was increased. Expression of transition protein (TP-l) and localization of peanut agglutinin were not seen until 3 weeks after culturing, and appeared by 6 weeks of culture. The putative spermatids derived from GSCs supported embryonic development up to the blastocyst stage with normal chromosomal ploidy after chemical activation. Thus, GSCs isolated from neonatal and adult mouse testes were able to be maintained and proliferated in our simple culture conditions. These GSCs have the potential to differentiate into haploid germ cells during another long-term culture.

• Korean Journal of Clinical Laboratory Science
• /
• v.41 no.3
• /
• pp.111-122
• /
• 2009

Cadmium (Cd) is known to exert gonadotoxic and spermiotoxic effects. The present study was performed to investigate the morphological effects and metallothionein (MT) expression by zinc pretreatment in the course of time of cadmium-induced testicular injury in rat. Fifty male Spraque-Dawley rats weighing 160~180 g were divided into two groups : saline-pretreated cadmium group and zinc-pretreated cadmium group. Rats of two groups received subcutaneous injection of saline and 100 mg/kg $ZnSO_4$ at 0, 2, 5 and 8 hrs intervals respectively. Cadmium chloride (4.5 mg/kg $CdCl_2$) was administrated intraperitoneally at 2 hrs after zinc injection and rats were killed 0, 12, 24, 48 and 72 hrs later. Testicular tissue damages, interstitial (Leydig) cells status and MT expression were determined using hematoxylin-eosin stained sections and a computerized image analysis system on sections immunostained with a mouse anti-metallothionein respectively. Zinc pretreatment was significantly reduced testicular damages in five pathological categories after cadmium administation. The number of surviving interstitial cells was significantly higher in the zinc-pretreated group than in the saline-preatreated group at 48 and 72 hrs after cadmium administration. Non-damaged testis showed the positivity of MT staining in spermatogenic cells, Sertoli cells and endothelium of blood vessel, but not in the Leydig cells. The positivity of MT staining in saline-pretreated group was significantly reduced at 24 hrs after cadmium administration, whereas zinc-pretreated group showed strong MT positive staining similar to the 0 hr by 42 hrs after cadmium administration. In damaged testis, MT positive staining was also observed in the Leydig cells of both groups. These results suggest that a major preventive effect of zinc against cadmium-induced testicular toxicity may be due to its ability to reduce the cytotoxicity of cadmium in spermatogenic cells and Leydig cells by inhibiting the susceptibility of the testis to cadmium but not MT production by cadmium.

• Korean Journal of Clinical Laboratory Science
• /
• v.42 no.3
• /
• pp.136-148
• /
• 2010

Cadmium (Cd) is known to exert gonadotoxic and spermiotoxic effects. The present study was performed to investigate the morphological effects and metallothionein (MT) expression by zinc pretreatment in the course of time of cadmium-induced testicular injury in rat. Fifty male Spraque-Dawley rats weighing 160-180 g were divided into two groups: saline-pretreated cadmium group and zinc-pretreated cadmium group. Rats of two groups received subcutaneous injection of saline and 100 mg/kg $ZnSO_4$ at 0, 2, 5 and 8 hrs intervals respectively. Cadmium chloride (4.5 mg/kg $CdCl_2$) was administrated intraperitoneally at 2 hr after zinc injection and rats were killed 0, 12, 24, 48 and 72 hrs later. Testicular tissue damages, Interstitial (Leydig) cells status and MT expression were determined using hematoxylin-eosin stained sections and a computerized image analysis system on sections immunostained with a mouse anti-metallothionein respectively. Zinc pretreatment was significantly reduced testicular damages in five pathological categories after cadmium administation. The number of surviving interstitial cells was significantly higher in the zinc-pretreated group than in the saline-preatreated group at 48 and 72 hrs after cadmium administration. Non-damaged testis showed the positivity of MT staining in spermatogenic cells, Sertoli cells and endothelium of blood vessel, but not in the Leydig cells. The potitivity of MT staining in saline-pretreated group was significantly reduced at 24 hrs after cadmium administration, whereas zinc-pretreated group showed strong MT positive staining similar to the 0 hr by 42 hrs after cadmium administration. In damaged testis, MT positive staining was also observed in the Leydig cells of both groups. These results suggest a major preventive effect of zinc against cadmium-induced testiculat toxicity may be due to its ability to reduce the cytotoxicity of cadmium in spermatogenic cells and Leydig cells by inhibiting the susceptibility of the testis to cadmium but not MT production by cadmium.

• Korean Journal of Animal Reproduction
• /
• v.11 no.1
• /
• pp.33-41
• /
• 1987

The cycle of the seminiferous epthelia in the testis of matured Korean Native Cattle was divided into eight stages. The results were summarized as follows: 1. Type A spermatogonia a, pp.ared twice as many at stage 2 as compared to stage 1, while maximum numbers were the average of 2.8 at stage 2. The intermediate and Type B spermatogonia were found during the stage 3 to 8, stage 6 to 8, respectively. The leptolene primary spermatocytes were not observed during the stage 5 to 7, while the pachytene primary spermatocytes were shown the least in number at stage 4, the secondary supermatocytes could be seen only at stage 4 and the round spermatids were not observed at stage 3, 4. 2. The relative frequencies of the eight stages of the cycle of the seminiferous eptithelia were 24.9, 14.2, 19.0, 6.3, 3.7, 7.9, 10.3 and 13.9%, respectively. 3. Some of the nuclei of Sertoli cells transformed from the "parallel" type to the "perpendicular" type. This evolution took place from stage 1 to 5, when the number of "perpendicular" type nuclei reached a peak and the number was decreased in the rest of the stages.sed in the rest of the stages.

• Korean Journal of Veterinary Research
• /
• v.33 no.1
• /
• pp.23-36
• /
• 1993

To investigate the cycle and relative frequences and the fine structure of seminiferous epithelia in mature Jindo dogs, histologic study was performed. The results obtained were summarized as follows; 1. Type A spermatogonia appeared approximately 1.6 times as many at stage II as compared to stage I while type In spermatogonia appeared small amount in stage III, IV and V. type B spermatogonia were found during the stage VI to VIII, though not detectable during stage I to V. The type B spermatogonia divided at stage VII to produce the preleptotene primary spermatocytes at stage VIII. The number of primary spermatocytes of the leptotene phase markedly increased during stage I to II, and the primary spermatocytes of the pachytene phase were shown the least in number at stage IV. The secondary spermatocytes could be seen only at stage IV. 2. The relative frequencies of each stage from stages I to VIII of the cycle of seminiferous epithelia were 31.6, 11.9, 10.0, 3.2, 8.2, 10.1, 11.7 and 13.2% respectively. 3. On electron microscopic observations, acrosomal vesicle of spermatids appeared larger though the bulk of germ cells were the morphologically same as those of the other animal species. Thread line structures light microscopically observed in the cytoplasm of Sertoli cell were the longitudinal orientation of mitochondria.

• Journal of radiological science and technology
• /
• v.13 no.2
• /
• pp.51-51
• /
• 1990

This study was undertaken to observe the effects of $^{60}Co\;{\gamma}-irradiation$ on the cell of spermatogenic epithelium in the testis of the chicken. 16-week-old chicken were provided as an experimental group and compared with control group. The experimental group was divided into a single irradiation (800, 1000, 1200 rads) and into three partial irradiation group (800/3, 1000/3, 1200/3 rads). The morphological changes of epithelial cell of the testis were observed by means of hematoxyline and eosin stain. Microstructure of spermatocyte and sperm was observed by means of semithin section of electron microscopic specimen. The results obstained are summerized as follows. 1. Spermatogonia and sertoli cells were found to be isolated from the basal membrane of seminiferous tubules as dose of $^{60}Co\;{\gamma}-irradiation$ was increased. 2. Spermatocytes of pachytene stage were seperated from the cytotplasmic process of sertoil cell in case of 1000 rads of $^{60}Co\;{\gamma}-irradiation$. 3. Normal arrangement of the cell of spermatogenic epithelium was found in control group and only the partial irradiation group of 800 rads. Vaculation in the seminiferous was pronounced in case of a single irradiation group of 800 rads, but the irradiation group of 1000 rads and 1200 rads were found to be damaged severely in both a single and a partial dose.

• Asian-Australasian Journal of Animal Sciences
• /
• v.22 no.2
• /
• pp.187-193
• /
• 2009

The present study identified the favorable conditions for isolation, enrichment and in vitro culture of highly purified, undifferentiated pig spermatogonial stem cell (SSC) lines that proliferate for long periods of time in culture. The colonies displayed morphology similar to miceSSC and were positive for markers of SSC (PGP9.5), proliferating germ cell (PigVASA), pre-meiotic germ cell (DAZL) and pluripotency (OCT4, SSEA-1, NANOG, and SOX2) based on immuno-cytochemistry and RT-PCR. The purity of these colonies was confirmed by negative expression of markers for sertoli cell (GATA4 and SOX9), peritubular myoid cell (${\alpha}$-SMA), differentiating spermatogonial and germ cells (c-KIT). The colonies could be maintained with undifferentiated morphology for more than two months and passaged more than 8 times with doubling time between 6-7 days. Taken together, we conclude that pigSSC could be successfully isolated and cultured in vitro and they possess characteristics similar to miceSSC.

• Clinical and Experimental Reproductive Medicine
• /
• v.26 no.2
• /
• pp.225-230
• /
• 1999

In mouse, the heat shock protein 70-2 (hsp70-2) is found to have special function in spermatogenesis. Based on the observation, the hypothesis that human hspA2 (human gene; 98.2% amino acid homology with hsp70-2) might have important function in spermatogenesis in human testes was proposed. To test the hypothesis, we examined the expression of hspA2 in human tissues. Expression vector pDMC4 for expression of the human hspA2 protein using pTricHisB (invitrogen, USA) was constructed and the expressed hspA2 protein was cross-reacted with antiserum 2A raised against mouse hsp70-2 protein. Based on the cross-reactivity, we determined the expression level of hspA2 protein in human tissues by western blot analysis using the antiserum 2A. We demonstrated that antiserum 2A antibodies detected human hspA2 protein with specificity which was produced in the E.coli expression system. On Western blot analyses, significant hspA2 expression was observed in testes with normal spermatogenesis, whereas a low level of hspA2 was expressed in testis with Sertoli-cell only syndrome. Also, a small amount of hspA2 was detected in breast, stomach, prostate, colon, liver, ovary, and epididymis. These results demonstrate that the hspA2 protein is highly expressed in male specific germ cells, which in turn suggests that hspA2 protein might playa specific role during meiosis in human testes as suggested in the murine model. However, further studies should be attempted to determine the function of hspA2 protein in human spermatogenesis.

• Journal of Embryo Transfer
• /
• v.28 no.2
• /
• pp.149-155
• /
• 2013

Mullerian inhibiting substance (MIS) is a member of the TGF-${\beta}$ (transforming growth factor-${\beta}$) family whose members play key roles in development, suppression of tumour growth, and feedback control of the pituitary-gonadal hormone axis. MIS is expressed in a highly tissue-specific manner in which it is restricted to male Sertoli cells and female granulose cells. The serum levels of MIS in prenatal and postnatal ICR mice were measured using the enzyme-linked immuno-solvent assay (ELISA) using the MIS/AMH antibody. Mice were grouped by age: the significant periods were at the onset of development. During sex organ differentiation, no remarkable difference between female and male foetus MIS serum levels (both<0.1 ng/ml) was observed. However, MIS serum levels in pregnant mice markedly changed (4.5~12.2 ng/ml). After birth, postnatal female and male mice serum MIS levels changed considerably (male: <0.1~138.5 ng/ml, female: 5.3~103.4 ng/ml), and the changing phase were diametrically opposed (male: decreasing, female: fluctuating). These findings suggest that MIS may have strong associations with not only develop-ment but also puberty. For further studies, establishing the standard MIS serum levels is of importance. Our study provides the basic information for the study of MIS interactions with reproductive organ disability, cancer, and the effect of other hormone or menopause. We hypothesise that if MIS is regularly injected into middle-age women, meno-pause will be delayed. We detected that serum MIS concentration curves change with age. The changing phase is different between males and females, and this difference is significant after birth. Moreover, MIS mRNA is expressed during the developmental period (prenatal) and also in the postnatal period. This finding indicates that MIS may play a significant role in the developmental stage and in growth after birth.