• Journal of Embryo Transfer
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• v.20 no.1
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• pp.35-41
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• 2005

This study was carried out for development of effective preservation on animal genetic resources. Spermatogonia cells are the germline stem cells and they can be restored to adult animal with proliferation and differentiation intentionally. When the spermatogonia cells were purified from seminiferous tubules and were cultured at $32^{\circ}C$, the cells were actively proliferated. The culture medium consisted of TCM199 plus $10\%$ FCS and coculture with Sertoli cells supported cultivation of spermatogonia cells. By passing 40 days of incubation, spermatogonia cells formed the germline colony or shape of ES-like colony or reconstruction of pseudo-seminifcrous tubule shape. At 40 days, the cultured cells were no sign for differentiation to spermatocyte or spermatid. The experiment of induced differentiation of this cells is needed.

• Korean Journal of Environmental Biology
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• v.22 no.4
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• pp.550-558
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• 2004

The effects of postnatal exposure to octylphenol(OP) on the expressions of the steroidogenic enzymes and testosterone production were evaluated. Postnatal male mice (15-day-old) were injected with 2 or 20mg $kg^{-l}$ body weight (BW) of OP for 5 days and sacrificed on postnatal day 21. Testosterone concentration was measured by radioimmunoassay and the expressions of the testicular genes were determined by RT-PCR analyses. Significant reductions in the mean body and testis weight were observed in the OP treated animals. No marked alteration in the histological structure of the testis were observed, however, slight reduction in the seminiferous tubule diameter and the number of Leydig cells and several pyknotic cells could be identified in the 20 mg $kg^{-l}$ BW of the OP treated animals. Serum testosterone concentration was dramatically reduced and the mRNA expressions of the steroidogenic acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (P450scc) and $17\beta$-hydroxylase/Cl7-20 lyase $(P450_{17\alpha})$ were decreased. No significant changes of the gene expressions of the steroidogenic factor-l (SF-I) and estrogen and androgen receptor after the OP treatment showed that the decreased expressions of the steroidogenic enzymes in the present study did not correlate with these genes. Altogether, the present study demonstrates that postnatal treatment of OP inhibits steroidogenesis by decreasing the transcriptional expressions of the StAR and steroidogenic enzymes. The alteration in steroidogenesis may adversely affect the normal development of the testis and sper- matogenesis.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.11
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• pp.1729-1737
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• 2018

Objective: The aim of the current study is to investigate the relationship between prohibitin (PHB), capping actin protein of muscle Z-line beta subunit (CAPZB), and tektin-2 (TEKT2) and sperm motility in Murrah buffalo. Methods: We collected the high-motility and low-motility semen samples, testis, ovary, muscle, kidney, liver, brain and pituitary from Murrah buffalo, and analysed the expression of PHB, CAPZB, and TEKT2 in mRNA (message RNA) and protein level. Results: Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) result showed that the expression of PHB was higher and CAPZB, TEKT2 were specifically expressed in testis as compared to the other 6 tissues, and that in testis, the expression of TEKT2 was higher than that of CAPZB and PHB. Immunohistochemistry test revealed that all three genes were located on the convoluted seminiferous tubule and enriched in spermatogenic cells. Both qRT-PCR and Western Blot results showed that the expression levels of PHB, CAPZB, and TEKT2 were significantly lower in the low-motility semen group compared to the high-motility semen group (p<0.05). Conclusion: The expression levels of PHB, CAPZB, and TEKT2 in Murrah buffalo sperm have a high positive correlation with sperm motility. And the three genes may be potential molecular markers for the decline of buffalo sperm motility.

• The Korean Journal of Zoology
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• v.33 no.1
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• pp.78-93
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• 1990

The ultrastructure of epididymal epithelium of 10, 20, 35 and 80 day-old mouse was observed to study the differentiation and function of the epithelial cells in connection with the absorption and secretion during sexual matruation. The differentiation of epididymis was divided into three phases of, 1) undiflerentiated phase until the day 20 after birth, 2) growing and differentiating phase between the day 20 and 35, and 3) maturating phase up to the adult. Each phase was closely related with the lumination of seminiferous tubule and the influx of spermatozoa within testicular fluid from testis. In adult, the ultrastructural features appeared an absorptive function in the principal cells of proximal caput epididymis, and a strong activity of protein synthesis and secretion in distal caput, corpus and cauda epididymis. Clear cells were predominantly located in corpus and cauda epidiymis, and plenty of absorption vesicles including membranous particles assumed to be the cellular residues from spermatozoa were observed at apical region. Therefore, the distribution of various cell types of epithelium and the ultrastructure even in the same type of epithelial cell, were different according to the epididymal regions.

• Development and Reproduction
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• v.23 no.2
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• pp.93-99
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• 2019

Nonylphenols (NPs) are widely used industrial materials, and are considered as potent endocrine disrupting chemical. Present study was undertaken to clarify the effect of subchronic low-dose NP exposure to F1 generation male mice. Mice were divided into 2 groups; (1) CON, control animals and (2) NP-50 ($50{\mu}g/L$), animals were treated with NP via drinking water. NP exposures were continuously conducted from parental pre-mating period until the postnatal day (PND) 55 of F1 offsprings. Mice were sacrificed on PND 55 and the tissue weights were measured. The initial body weights (at PND 21) and terminal body weights (PND 55) of the NP-50 animals were significantly lower than those of control animals (p<0.05). NP exposure induced a significant increase in the absolute weight of the testes (p<0.05). Conversely, the NP exposure caused significant decrease in the absolute weights of the epididymis (p<0.01), prostate (p<0.05) and seminal vesicle (p<0.05). Histopathological studies revealed that NP-treated animals exerted decreased seminiferous tubule diameters, reduced luminal area, and lower number of germ cells. Also some sloughing morphologies in the tubules were observed. In the caudal epididymis, fewer mature sperms and swollen epithelial cells were found in the NP-treated group. Our results confirmed that the subchronic low-dose NP exposure altered some male parameters and induced histopathological abnormalities in testis and epididymis of F1 mice. Since the NP dose used in this study is close to the average human daily NP exposure, our results could provide practically meaningful understanding of adverse effect of EDC in human.

• Development and Reproduction
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• v.23 no.3
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• pp.255-262
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• 2019

Previously, we reported negative effects of low-dose nonylphenol (NP) exposure on the reproductive organs of F1 male mice. In the present study was further investigated the endocrine disrupting effect of NP exposure to F2 generation male mice. Mice were divided into 2 groups; (1) CON, control animals and (2) NP-50 ($50{\mu}g/L$), animals were treated with NP via drinking water. NP exposures were continuously conducted from parental pre-mating period until the postnatal day (PND) 55 of F2 offsprings. Mice were sacrificed on PND 55 and the reproductive tissue weights were measured. The initial (at PND 21) and terminal (PND 55) body weights of the NP-50 group animals were not significantly different from those of control group animals. NP exposure fail to induce a significant weight change of the testes, seminal vesicle and prostate except absolute epididymal weight (p<0.05). However, pathohistological studies revealed that NP-treated F2 animals showed evident decrease in seminiferous tubule diameters, reduced luminal area and number of germ cells. Also, sloughing morphologies in the tubules were notable. In the caudal epididymis, fewer mature sperms and swollen epithelial cells were found in the NP-treated group. The present study demonstrated that the subchronic low-dose NP exposure induced pathohistological abnormalities in testis and epididymis of F2 mice, and we assumed that these 'qualitative' changes in reproductive tissues could be derived from the epigenetic modifications such as DNA methylation, histone modification, altered DNA accessibility and chromatin structure. Further studies are needed to achieve a better understanding on the multi- or trans-generational effects of NP on the reproductive health and a human application.

• Clinical and Experimental Reproductive Medicine
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• v.50 no.4
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• pp.230-243
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• 2023

Objective: High temperatures can trigger cellular oxidative stress and disrupt spermatogenesis, potentially leading to male infertility. We investigated the effects of retinoic acid (RA), chitosan nanoparticles (CHNPs), and retinoic acid loaded with chitosan nanoparticles (RACHNPs) on spermatogenesis in mice induced by scrotal hyperthermia (Hyp). Methods: Thirty mice (weighing 25 to 30 g) were divided into five experimental groups of six mice each. The groups were as follows: control, Hyp induced by a water bath (43 ℃C for 30 minutes/day for 5 weeks), Hyp+RA (2 mg/kg/day), Hyp+CHNPs (2 mg/kg/72 hours), and Hyp+RACHNPs (4 mg/kg/72 hours). The mice were treated for 35 days. After the experimental treatments, the animals were euthanized. Sperm samples were collected for analysis of sperm parameters, and blood serum was isolated for testosterone measurement. Testis samples were also collected for histopathology assessment, reactive oxygen species (ROS) evaluation, and RNA extraction, which was done to compare the expression levels of the bax, bcl2, p53, Fas, and FasL genes among groups. Additionally, immunohistochemical staining was performed. Results: Treatment with RACHNPs significantly increased stereological parameters such as testicular volume, seminiferous tubule length, and testicular cell count. Additionally, it increased testosterone concentration and improved sperm parameters. We observed significant decreases in ROS production and caspase-3 immunostaining in the RACHNP group. Moreover, the expression levels of bax, p53, Fas, and FasL significantly decreased in the groups treated with RACHNPs and RA. Conclusion: RACHNPs can be considered a potent antioxidative and antiapoptotic agent for therapeutic strategies in reproductive and regenerative medicine.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.10 no.2
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• pp.71-96
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• 1977

Early development Linuparus trigonus(von Siebold) has been studied based on the samples collected monthly in Je-ju Island, Korea from February, 1975 to January, 1977. Gametogenesis, reproductive cycle, embryonic development were investigated by histological mettled, and morphological description was made on the first phyllosoma larva which reared in the laboratory. Testis is composed of two tubular duct which are symmetrical with H-shaped appearance. Outer layer of testis is of fibrous connective tissue capsule. In the lumen there is a convoluted seminiferous tubule with interstitial tissue. Ovary is a pair of symmetrical blind tubular lobes, and the midportions are connected each other. The ovary consists of a couple of ovarian sacs partitioned by two-layered connective tissue fibers. Proliferation of spermatogonia are observed all the year around on the germinal epithelium of seminiferous tubule. Partial spermatogenesis is always in progress, and the spermatozoa appear all the year around in the tubules. Nutrition of early oogonia is supplied by fibrous mesenchyme which is abundantly distributed in ovarian sacs. Oocytes grow and couplete maturation divisions in the follicle layers. They finally develop into mature ova before spawning. Reproductive cycle is classified into four successive stages; multiplication stage from September to December, growing stage from January to March, maturation division stage from April to May and mature stage from June to August. Spawning takes place from May to August with peak spawning from Into July to early August. Cleavage type is superficial. Blastopore is formed in blasto-disc region which is proliferation of blastoderm cells. Germinal layers are also derived from tile region. Mesoderm formation is originated from endodermal cells which are formed front the blasto-disc region. The endodermal cells are separated by the process of delamination from yolk sac and take part in the formation of the mid-gut. Morphological characteristics of first phyllosoma larva are different from the larvae of other Palinurid and Scyllarid species.

• Development and Reproduction
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• v.11 no.1
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• pp.31-41
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• 2007

The spermiogenesis of Crocidura shantungensis were studied by electron microscope. All process of spermiogenesis was divided into 11 phases 15 steps, based on the morphological features of the nucleus and cell organelles in cytoplasm of spermatids. The spermatids in Golgi and cap phases were a spherical shape. On the other hand, at the early acrosomal phase they changed into an oval shape, and the tail was created in this phase. In maturation phase, the shapes of spermatid head were thin and longish. Until step 7 the direction of spermatids head turned toward the lumen of the seminiferous tubule. From step 8 to step 15 their heads turned toward the basal lamina. In step 12, the nucleus and acrosome shown maximal elongation. From Step 13 the nucleus of spermatids became flat, simultaneously with flat expansion of the acrosome expanded, and the visible whole lengths of spermatids were tend to be shorten. Spermatid heading which arrived to step 14 was taken the final shape. The nucleus was doing the wedge shape, and the nuclear chromatins condensed completely and homogenized. In the spermiation phase, the spermatids were gradually disconnected from the cytoplasm of the Sertoli cell. In this phase, the acrosome of the spermatids were fully shorten and flat, and the spermatozoa completed the process of heading and the tailing. Considering all the results, the spermiogenesis may be useful information to analyze the differentiation of spermatogenic cells.

• BMB Reports
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• v.38 no.6
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• pp.739-747
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• 2005

Full-length cDNA sequences of four novel SPATA4 genes in chimpanzee, cow, chicken and ascidian were identified by bioinformatic analysis using mouse or human SPATA4 cDNA fragment as electronic probe. All these genes have 6 exons and have similar protein molecular weight and do not localize in sex chromosome. The mouse SPATA4 sequence is identified as significantly changed in cryptorchidism, which shares no significant homology with any known protein in swissprot databases except for the homologous genes in various vertebrates. Our searching results showed that all SPATA4 proteins have a putative conserved domain DUF1042. The percentages of putative SPATA4 protein sequence identity ranging from 30% to 99%. The high similarity was also found in 1 kb promoter regions of human, mouse and rat SPATA4 gene. The similarities of the sequences upstream of SPATA4 promoter also have a high proportion. The results of searching SymAtlas (http://symatlas.gnf.org/SymAtlas/) showed that human SPATA4 has a high expression in testis, especially in testis interstitial, leydig cell, seminiferous tubule and germ cell. Mouse SPATA4 was observed exclusively in adult mouse testis and almost no signal was detected in other tissues. The pI values of the protein are negative, ranging from 9.44 to 10.15. The subcellular location of the protein is usually in the nucleus. And the signal peptide possibilities for SPATA4 are always zero. Using the SNPs data in NCBI, we found 33 SNPs in human SPATA4 gene genomic DNA region, with the distribution of 29 SNPs in the introns. CpG island searching gives the data about CpG island, which shows that the regions of the CpG island have a high similarity with each other, though the length of the CpG island is different from each other.This research is a fundamental work in the fields of the bioinformational analysis, and also put forward a new way for the bioinformatic analysis of other genes.