• Asian-Australasian Journal of Animal Sciences
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• v.31 no.6
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• pp.791-797
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• 2018

Objective: Trimethylation of histone 3 (H3) at 4th lysine N-termini (H3K4me3) in gene promoter region was the universal marker of active genes specific to cell lineage. On the contrary, coexistence of trimethylation at 27th lysine (H3K27me3) in the same loci-the bivalent H3K4m3/H3K27me3 was known to suspend the gene transcription in germ cells, and could also be inherited to the developed stem cell. In galline species, throughout example of H3K4m3 and H3K27me3 ChIP-seq analysis was still not provided. We therefore designed and demonstrated such procedures using ChIP-seq and mRNA-seq data of chicken follicular mesenchymal cells and male germ cells. Methods: Analytical workflow was designed and provided in this study. ChIP-seq and RNA-seq datasets of follicular mesenchymal cells and male germ cells were acquired and properly preprocessed. Peak calling by Model-based analysis of ChIP-seq 2 was performed to identify H3K4m3 or H3K27me3 enriched regions ($Fold-change{\geq}2$, $FDR{\leq}0.01$) in gene promoter regions. Integrative genomics viewer was utilized for cellular retinoic acid binding protein 1 (CRABP1), growth differentiation factor 10 (GDF10), and gremlin 1 (GREM1) gene explorations. Results: The acquired results indicated that follicular mesenchymal cells and germ cells shared several unique gene promoter regions enriched with H3K4me3 (5,704 peaks) and also unique regions of bivalent H3K4m3/H3K27me3 shared between all cell types and germ cells (1,909 peaks). Subsequent observation of follicular mesenchyme-specific genes-CRABP1, GDF10, and GREM1 correctly revealed vigorous transcriptions of these genes in follicular mesenchymal cells. As expected, bivalent H3K4m3/H3K27me3 pattern was manifested in gene promoter regions of germ cells, and thus suspended their transcriptions. Conclusion: According the results, an example of chicken H3K4m3/H3K27me3 ChIP-seq data analysis was successfully demonstrated in this study. Hopefully, the provided methodology should hereby be useful for galline ChIP-seq data analysis in the future.

• Tuberculosis and Respiratory Diseases
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• v.58 no.2
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• pp.184-187
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• 2005

The Mixed germ cell tumors of the mediastinum are very quite rare. The Prognosis is generally dominated by the most aggressive component, which is represented by a choriocarcinoma, an endodermal sinus tumor, an embryonal carcinoma, and a seminoma, in descending order of in the degree of malignancy. We experienced one a case of a mixed germ cell tumor at the anterior mediastinum. The patient was 27-year-old male, who complained of hemoptysis and cough. The Chest X-ray showed a well-defined lobulated mediastinal mass in the left upper lung field. The operation was done and The mass was excised surgically. A Biopsy showed elements of mature tissues, immature neuronal components, and seminoma components.

• Tuberculosis and Respiratory Diseases
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• v.47 no.1
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• pp.117-122
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• 1999

Primary germ cell tumors of the mediastinum are rare, accounting 1-5 % among all germ cell tumors and 10% of all neoplasms in this area. Approximately 85 % of these tumors occur in men with a mean age 29 years. 'These tumors are mainly found in the anterior mediastinum and appear grossly as large lobulated masses. They are frequently invasive at the time of diagnosis and almost 90% of patients are symptomatic. Primary nonseminomatous germ cell tumor arising in the posterior mediastinum is very rare. We report a case of 37-year old male arising from the posterior mediastinum. Serum tumors markers including alpha-fetoprotein and $\beta$-hCG which are usually elevated in germ cell tumor were not elevated. He was found to have a primary mediastinal embryonal carcinoma with pulmonary metastasis at open exploration. He was treated with debulking surgery and cisplatin-based chemotherapy, died of sepsis after 15 months postoperatively.

• Reproductive and Developmental Biology
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• v.31 no.2
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• pp.127-131
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• 2007

Embryonic germ (EG) cells are undifferentiated stern cells isolated from cultured primordial germ cells (PGC). These cells share many characteristics with embryonic stem cells including morphology and pluripotency. Undifferentiated porcine EG cell lines demonstrating capacities of differentiation both in vitro and in vivo have been established. Since EG cells can be cultured indefinitely in an undifferentiated state, whereas somatic cells in primary culture are often unstable and have limited lifespan, EG cells may provide inexhaustible source of karyoplasts in nuclear transfer (NT). In this study the efficiencies of NT using porcine EG and fetal fibroblast cells were compared. Two different techniques were used to perform NT. With conventional NT procedure (Roslin method) involving fusion of donor cells with enucleated oocytes, the rates of development to the blastocyst stage in EG and somatic cell NT were 16.8% (59/351) and 14.5% (98/677), respectively. In piezo-driven microinjection (Honolulu method) of donor nuclei into enucleated oocytes, the rates of blastocyst formation in EG and somatic cell NT were 11.9% (15/126) and 9.4% (9/96), respectively. Regardless of NT methods used in this study, EG cell NT gave rise to comparable rate of blastocyst development to somatic cell NT. Overall, EG cells can be used as karyoplast donor in NT procedure, and embryos can be produced by EG cell NT that may be used as an alternative to conventional somatic cell NT.

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

• Development and Reproduction
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• v.20 no.1
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• pp.11-22
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• 2016

The ultrastructures of germ cells and the functions of Leydig cells and Sertoli cells during spermatogenesis in male Kareius bicoloratus (Pleuronectidae) were investigated by electron microscope observation. Each of the well-developed Leydig cells during active maturation division and before spermiation contained an ovoid vesicular nucleus, a number of smooth endoplasmic reticula, well-developed tubular or vesicular mitochondrial cristae, and several lipid droplets in the cytoplasm. It is assumed that Leydig cells are typical steroidogenic cells showing cytological characteristics associated with male steroidogenesis. No cyclic structural changes in the Leydig cells were observed through the year. However, although no clear evidence of steroidogenesis or of any transfer of nutrients from the Sertoli cells to spermatogenic cells was observed, cyclic structural changes in the Sertoli cells were observed over the year. During the period of undischarged germ cell degeneration after spermiation, the Sertoli cells evidenced a lysosomal system associated with phagocytic function in the seminiferous lobules. In this study, the Sertoli cells function in phagocytosis and the resorption of products originating from degenerating spermatids and spermatozoa after spermiation. The spermatozoon lacks an acrosome, as have been shown in all teleost fish spermatozoa. The flagellum or sperm tail of this species evidences the typical 9+2 array of microtubules.

• Journal of Animal Science and Technology
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• v.64 no.4
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• pp.654-670
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• 2022

Spermatogenesis and testis development are highly structured physiological processes responsible for post-pubertal fertility in stallions. Spermatogenesis comprises spermatocytogenesis, meiosis, and spermiogenesis. Although germ cell degeneration is a continuous process, its effects are more pronounced during spermatocytogenesis and meiosis. The productivity and efficiency of spermatogenesis are directly linked to pubertal development, degenerated germ cell populations, aging, nutrition, and season of the year in stallions. The multiplex interplay of germ cells with somatic cells, endocrine and paracrine factors, growth factors, and signaling molecules contributes to the regulation of spermatogenesis. A cell-tocell communication within the testes of these factors is a fundamental requirement of normal spermatogenesis. A noteworthy development has been made recently on discovering the effects of different somatic cells including Leydig, Sertoli, and peritubular myoid cells on manipulation the fate of spermatogonial stem cells. In this review, we discuss the self-renewal, differentiation, and apoptotic roles of somatic cells and the relationship between somatic and germ cells during normal spermatogenesis. We also summarize the roles of different growth factors, their paracrine/endocrine/autocrine pathways, and the different cytokines associated with spermatogenesis. Furthermore, we highlight important matters for further studies on the regulation of spermatogenesis. This review presents an insight into the mechanism of spermatogenesis, and helpful in developing better understanding of the functions of somatic cells, particularly in stallions and would offer new research goals for developing curative techniques to address infertility/subfertility in stallions.

• The Korean Journal of Malacology
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• v.27 no.2
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• pp.121-129
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• 2011

The ultrastructural characteristics of germ cell development during spermatogenesis and mature sperm morphology of in male Scapharca subcrenata were investigated by transmission electron microscope observation. Spermatogonia are located nearest the outer wall of the acinus, while spermatocytes and spermatids are positioned near the accessory cells. The accessory cells, which is in close contact with developing germ cells, contained a large quantity of glycogen particles and lipid droplets in the cytoplasm. Therefore, it is assumed that they are involved in supplying of the nutrients for germ cell development. The morphologies of the sperm nucleus and the acrosome of this species are the oval shape and cone shape, respectively. Spermatozoa are approximately 45-$50{\mu}m$ in length including a sperm nucleus (about $1.30{\mu}m$ in length), an acrosome (about $0.59{\mu}m$ in length), and tail flagellum (about 43-$47{\mu}m$). The axoneme of the sperm tail shows a 9 + 2 structure. As some characteristics of the acrosomal vesicle structures, the right and left basal rings show electron opaque part (region), and also the anterior apex part of the acrosomal vesicle shows electron opaque part (region). These characteristics of the acrosomal vesicle were found in Acinidae and other several families in subclass Pteriomorphia. These common characteristics of the acrosomal vesicle in subclass Pteriomorphia can be used for phylogenetic and taxonomic analysis as a taxonomic key or a significant tool. The number of mitochondria in the midpiece of the sperm of this species are five, as one of common characteristics appear in most species in Arcidae and other families in subclass Pteriomorphia. The acrosomal vesicles of Arcidae species do not contain the axial rod and several transverse bands in acrosome, unlkely as seen in Ostreidae species in subclass Pteriomorphia, These characteristics can be used for the taxonomic analysis of the family or superfamily levels as a systematic key or tools.

• Journal of Animal Science and Technology
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• v.49 no.2
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• pp.183-194
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• 2007

We examined the expression patterns of the chicken TCs(tentative consensus sequences) originated from GermOnline genes in various chicken tissues, applying information from GermOnline to chicken organisms. 42 TCs among 84 chicken homologous TCs from the pool of 84 genes related to germ cell lineage in mouse(10), rat(71) and human(3) had high homology based on a BLAST search. Of these, Hmgcs2 and Sycp3 was shown to be expressed in a testis- specific manner and a reproductive organ(testis and ovary)-specific manner, respectively, by RT- PCR analysis. Crmp4, Cyct, Ldhc, Epha7, Pcsk4 and Dnmt3a are expressed in brain, testis, and ovary. The characterization of chicken genes originated from GermOnline in this research may give an enormously useful source of information related to germ cell development.

• Journal of electromagnetic engineering and science
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• v.15 no.3
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• pp.142-150
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• 2015

Several reports supported that continuous exposure to 60 Hz magnetic field (MF) induces testicular germ cell apoptosis in vivo. We recently evaluated duration- and dose-dependent effects of continuous exposure to a 60 Hz MF on the testes in mice. BALB/c male mice were exposed to a 60 Hz MF at $100{\mu}T$ for 24 hours a day for 2, 4, 6, or 8 weeks, and at 2, 20 or $200{\mu}T$ for 24 hours a day for 8 weeks. To induce the apoptosis of testicular germ cell in mice, the minimum dose is $20{\mu}T$ at continuous exposure to a 60 Hz MF for 8 weeks, and the minimum duration is 6 weeks at continuous exposure of $100{\mu}T$. Continuous exposure to a 60 Hz MF might affect duration- and dose-dependent biological processes including apoptotic cell death and spermatogenesis in the male reproductive system of mice. The safety guideline of the International Commission on Non-Ionizing Radiation Protection (ICNIRP) indicates that the permissible maximum magnetic flux density for general public exposure is $200{\mu}T$ at 60 Hz EMF (ICNIRP Guidelines, 2010). In the present study, we aimed to examine the expression of pro- and anti-apoptotic genes regulated by the continuous exposure to 60 Hz at $200{\mu}T$ in Sprague-Dawley rats for 20 weeks. The continuous exposure to 60 Hz at $200{\mu}T$ does not affect the body and testicular weight in rats. However, exposure to 60 Hz MF significantly affects testicular germ cell apoptosis and sperm count. Further, the apoptosis-related gene was scrutinized after exposure to 60 Hz at $200{\mu}T$ for 20 weeks. We found that the message level of endonuclease G (EndoG) was greatly increased following the exposure to 60 Hz at $200{\mu}T$ compared with sham control. These data suggested that 60 Hz magnetic field induced testicular germ cell apoptosis through mitochondrial protein Endo G.