Micro-deletions at specific loci of the Y chromosome have been observed frequently in male infertility patients, suggesting that genes in these regions are involved in male germ cell development. DAZ is a representative male infertility gene at the AZFc locus of the Y chromosome. Since DAZ contains an RNA binding motif along with so-called a DAZ domain, it was proposed to participate in RNA metabolism during spermatogenesis. A mouse gene homologous to the human DAZ gene has been cloned and named Dazl (DAZlike). Dazl is autosomal and expressed in the testis and also at a low level in the ovary. Male mice homozygous for the Dazl null allele have small testes with a few spermatogonia and almost complete absence of germ cells beyond the spermatogonial stage, suggesting the requirement of Dazl for entry or progression through meiosis. However, its exact cellular functions have not been understood yet. In order to investigate cellular functions of Dazl, we decided to isolate candidate interacting protein genes of the mouse Dazl, using yeast two-hybrid screening. A number of candidate Dazlinteracting proteins have been isolated, such as Bprp, Acf, Hgs, Murr1, Nbak3 and Ranbp9, but dynein light chain 1 (Dlc1) was most predominant. A strong interaction of Dazl with Dlc1 suggests that Dazl might function as an mRNA adaptor to the dynein motor complex.
Purpose : This study was conducted to investigate the effects of Cynomorii Herba (鎖陽) on the reproductive competence in male mice. Methods : We used 8-week-old Spraque-Dawley rats and administered the extract powder of Cynomorii Herba to 5 rats and normal saline to control group once in a day for 28 days. Then we observed the changes of body weight before and after administration of Cynomorii Herba extracts and normal saline. We isolated their testis surgically and observed the weight of testis, epididymis, vascular gland and prostate. Also we examined the total, normal and motile sperm concentration, the concentration of testicular catalase, peroxidase and configuration of testicular tissue before and after administration of Cynomorii Herba extracts and normal saline. Results : We found that the concentration of total, normal, motile sperm in testis of Cynomorii Herba treated group shows significant difference compared with the control group. The body weight, the weight of testis, epididymis and the concentration of testicular catalase and peroxidase were higher in the Cynomorii Herba treated group, on the other hand the weight of vascular gland was lower in the Cynomorii Herba. But the contents of above statement showed no significant difference. The weight of prostate showed lower in the control group statistically than that of the Cynomorii Herba treated group. We observed the seminiferous tubules taken shape minutely and the number of normal sperm increased in Cynomorii Herba treated group's testicular tissue. Conclusion This study shows that Cynomorii Herba may have an effect on the concentration, morphology and motility of sperm, the important factor in male fertility.
Gametogenesis, mating behaviour and spawning of Octopus ocellatus were investigated by histological study. This species is dioecious, and showed a protandry phenomenon. Ooogenesis (in females) and spermatogenesis (in males) can be classified into 3 stages, respectively. O. ocellatus copulates in one of two ways: a male may leap upon a female, mounting her mantle, or a male may sit near the female and extend the hectocotylized third right arm toward her. Spawning occurred between April and June in females, and between March and May in males of O. ocellatus. The spawning period was once a year and the peak took place between May and June. A number of flatened follicle cells, which were attached to an oocyte, were involved in vitellogenesis in the cytoplasm of the vitellogenic oocyte (maturing oocyte), and formation of chorion membrane (secondary egg membrane) of the ovarian eggs. Fecundity per female closely related to GSI was 294-660 eggs (average, 429 eggs). The diameters of the ovarian eggs surrounded by chorion membrane were approximately in the range of 10.10-2.50 mm. Each ovarian egg laid by a female was connected to an egg string. Each egg string was 1-5.5 cm (average 3.6 cm). The total number of eggs laid by a female of this species ranged 218-314, the egg sizes were independent to the size of female adult. this species has a life mode showing some special reproductive characteristics of an annual semelparity as shown in Octopodidae species because we have never seen a female spawning a second time.
19-norandrostenedione(19-NORA) is known as an intermediate in the metabolic pathway from androstenedione to estrone. Administration of esterified 19-nortestosterone, anabolic steroid, reduces serum gonadotropin and testosterone concentration, and results in reversible azoospermia in men. 19-NORA have been isolated from testis, but its function in testis is not clear yet. Therefore, this study was designed to determine the effect of 19-NORA on steroidogenesis and on spermatogenesis. 19-NORA was administrated by single intratesticular injection to adult male rats weighing 350-400 g in dose of 1 mg/50${\mu}l$. The serum and testis were collected on 1, 3, 7, 12, 48 hr after injection. The histological differences in testis were observed by routine paraffin method. The concentrations of testosterone and estradiol in serum and in left testis were determined by the conventional radioimmunoassays. One hour after 19-NORA treatment, serum concentrations of testosterone and estradiol increased significantly, compared to those of pre-treated(0 hr) group, and reduced gradually to the control level on 7 hour after injection. The concentration of testosterone in left testis increased slightly 1 hour after injection, and estradiol level increased significantly(p<0.05). Also, testosterone and estradiol level of control group revealed no difference with pre-treated (0 hr) group. Gonad index, structure of seminiferous tubules, and the number of step 7 th spermatid were simillar to control group. The present study suggests that the elevation of testosterone level results from increment of estradiol followed by the rapid metabolism of 19-NORA at 7 hour after injection, and then testosterone concentration may be recovered to control level by feedback mechanism of hypothalamus-hypothysis-testis axis.
Objective: In this study, specimens from testicular biopsies of men with nonobstructive azoospermia (NOA) were used to investigate whether RNF8 gene could serve as a biomarker to predict the presence of sperm in these patients. Methods: Testicular biopsy specimens from 47 patients were classified according to the presence of sperm (positive vs. negative groups) and investigated for the expression of RNF8. The level of RNF8 gene expression in the testes was compared between these groups using reverse-transcription polymerase chain reaction. Results: The expression level of RNF8 was significantly higher in testicular samples from the positive group than in those from the negative group. Moreover, the area under the curve of RNF8 expression for the entire study population was 0.84, showing the discriminatory power of RNF8 expression in differentiating between the positive and negative groups of men with NOA. A receiver operating characteristic curve analysis showed that RNF8 expression had a sensitivity of 81% and a specificity of 84%, with a cutoff level of 1.76. Conclusion: This study points out a significant association between the expression of RNF8 and the presence of sperm in NOA patients, which suggests that quantified RNF8 expression in testicular biopsy samples may be a valuable biomarker for predicting the presence of spermatozoa in biopsy samples.
Rezaei, Ali Akbar;Salehi, Iraj;Karimi, Seyed Asaad;Rahnama, Mehdi
Clinical and Experimental Reproductive Medicine
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제47권1호
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pp.34-41
/
2020
Objective: The strong antioxidant activity of Commiphora mukul prompted us to conduct the present study to explore whether treatment with C. mukul extract (CME) would have any protective influence on sperm parameters, testosterone levels, and plasma glucose levels in streptozotocin (STZ)-induced diabetic rats. Methods: Male Wistar rats were randomly divided into four groups: control, control animals treated with CME, diabetic animals, and diabetic animals treated with CME. CME extract (300 mg/kg) was administered for 60 days by daily gavage. Diabetes was induced by an intraperitoneal injection of 50 mg/kg STZ. The epididymal sperm count, weight, motility, morphology, viability, and serum testosterone and glucose levels were determined. Results: In the diabetic animals, CME decreased blood glucose levels (p< 0.05), increased the total sperm count (p< 0.05), and decreased the proportion of sperm with abnormal morphology (p< 0.05). Diabetes reduced sperm motility (p< 0.001), and CME supplementation partially reversed this effect of diabetes (p= 0.003). Furthermore, in diabetic animals, CME decreased the proportion of immotile sperm (p< 0.001). In rats, diabetes caused a significant decrease (p< 0.05) in serum testosterone levels (F[3, 28] = 3.283, p= 0.035), but treatment of diabetic animals with CME increased serum testosterone levels. Conclusion: The present study demonstrated that C. mukul possesses proandrogenic activity and exerts a beneficial effect on sperm parameters in diabetic rats.
Lee, Seung-Won;Wu, Guangming;Choi, Na Young;Lee, Hye Jeong;Bang, Jin Seok;Lee, Yukyeong;Lee, Minseong;Ko, Kisung;Scholer, Hans R.;Ko, Kinarm
Molecules and Cells
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제41권7호
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pp.631-638
/
2018
Spermatogonial stem cells (SSCs) derived from mouse testis are unipotent in regard of spermatogenesis. Our previous study demonstrated that SSCs can be fully reprogrammed into pluripotent stem cells, so called germline-derived pluripotent stem cells (gPS cells), on feeder cells (mouse embryonic fibroblasts), which supports SSC proliferation and induction of pluripotency. Because of an uncontrollable microenvironment caused by interactions with feeder cells, feeder-based SSC reprogramming is not suitable for elucidation of the self-reprogramming mechanism by which SSCs are converted into pluripotent stem cells. Recently, we have established a Matrigel-based SSC expansion culture system that allows longterm SSC proliferation without mouse embryonic fibroblast support. In this study, we developed a new feeder-free SSC self-reprogramming protocol based on the Matrigel-based culture system. The gPS cells generated using a feeder-free reprogramming system showed pluripotency at the molecular and cellular levels. The differentiation potential of gPS cells was confirmed in vitro and in vivo. Our study shows for the first time that the induction of SSC pluripotency can be achieved without feeder cells. The newly developed feeder-free self-reprogramming system could be a useful tool to reveal the mechanism by which unipotent cells are self-reprogrammed into pluripotent stem cells.
Jeong, Dong Kee;Sharma, Neelesh;Nguyen, Thanh Luan;Kim, Jong Hyun;Oh, Sung Jong
한국수정란이식학회지
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제29권4호
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pp.351-359
/
2014
Phasianus colchicus is not only a beautiful bird but also a great value in science and under the threat of endanger. Hence, the aim of this study was to isolate the pheasant male germ cells (mGCs) and then induce them into elongated sperm-like cells in vitro. The mGCs were purified and enriched by a two-step plating method based on the different adherence velocities of mGCs and somatic cells. The percentage of the c-kit positive cells and c-kit negative cells examined by flow cytometry analysis (FCA) was 92.87% and 2.57%, respectively. Subsequently, the mGCs were induced for 48h in DMEM/F12 medium supplemented factors such as retinol acid, testosterone and bovine FSH, followed by 5 weeks in culture. We found that some elongated sperm-like cells appeared initially in vitro under inducement of stimulated factors. The elongated sperm-like cells showed in the expression of changed morphology and post-transcriptional marker such as spermatid associated (SPERT), spermatid perinuclear RNA binding protein (STRBP), round spermatid basic protein 1 (RSBN1) and SPER1L. Moreover, in DNA content identified assay, induced cells showed that the 1C DNA population markedly increased in differentiated group but it was not change in undifferentiated group. Successful in vitro differentiation of pheasant testicular germline cells into spermatids appears to offer extremely attractive potential for the conservation of endangered birds and treatment of male infertility.
Objective: This study was undertaken to evaluate the dose dependent effects of Epimedii Herba extract solution on the spermatogenic abilities such as concentration, motility and morphological normality of sperm from the testis, and the activities of spermhyaluronidase and antioxidants. Materials and Method: We choose the 2-month-old mice, and administered the extract powder of Epimedii Herba in the different concentration once in a day for 60 days. The control group was administerde to normal water in the isolated testis tissue. Also we observed changes of isolated testis at the before and after administration of Epimedii Herba extracts in the mice. And we compared to the testicular tissue especially seminiferous tubules between control and treated group by histochemical methods. Results: The significant dose dependent differences were observed in the concentration of total sperm, the motility and normality of spermatozoa of the Epimedii Herba extract administered groups compared than that of control group, respectively. In the histological analysis of the testicular lobes were observed in the Epimedii Herbaextract administered groups than control group, respectively. Also, the activity hyluronidase was significantly increased in the Epimedii Herba extract administered groups than that of the control group. In case of antioxidant activity analysis, the activity of peroxidase and catalase were significantly increased in the Epimedii Herba extract administered groups than that of control group, respectively. Conclusion: This study shows that Epimedii Herba can effect the count and motility of sperm, the important ractor in male fertility and also promote the activity of antioxidants, catalase and peroxidase, which is the important factor in spermatogenesis.
To know the effect of Ivermectin(IVM) toxicity in testis, histopathologic changes as well as clinical signs were observed in experimental animals including dogs by the subcutaneous injection with 3-50mg/kg of IVM. Clinically, it was observed to have depression and ataxia in all groups whereas tremor and coma in mice, rats and guinea pigs, coma in hamsters and rabbits, and tremor and salivation in dogs were shown. The clinical signs were different by the dosage of IVM, species and individuals in all animals. Susceptibility to IVM was most sensitive in dogs, especially in a Tosa dog and this was susceptible in mice, hamsters and rabbits, guinea pigs and rats in order. Microscopical observation revealed that the seminiferous tubules of testis had decreased thickness of germinal epithelium due to the necrosis and desquamation of the spermatids and spermatocytes. The progressive pattern by the times of administration showed vacuolar formation between the layer of spermatids and spermatogonia due to the marked necrosis of spermatocytes and the presence of multinucleated giant cells derived from spermatid throughout the seminiferous tubules of testis. Only a layer of spermatogonia, a few spermatogonia, and Sertoli cells wore observed with atrophied wavelike basement membrane in the seminiferous tubules of testis. Necrotic germinal cells, sloughed immature spermatids and spermatocytes were present in the lumen of epididymis and ductus deferens. Microscopical observation showed different susceptibility to IVM with clinical observation in which it was also most sensitive in dogs, especially in a Tosa dog and this was susceptible in rabbits and guinea pigs, hamsters, rats and mice in order. It was considered that IVM affects mainly spermatocyte or spermatid stage in the spermatogenesis and disturbs their developing beyond these stage.
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