• Title/Summary/Keyword: Germ cell development

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Simple Classification of Male Mouse Germ Cells using Hoechst 33258 Staining (Hoechst 33258 Staining을 이용한 웅성 생쥐 성세포의 간편 분류)

  • Kim, Kyoung Guk;Park, Young Sik
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.213-218
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    • 2015
  • In the study for a differentiation and development of spermatogonial cells, the researchers should commonly require a simple, fast and reasonable method that could evaluate the developmental stage of male germ cells without any damage and also relentlessly culture them so far as a cell stage aiming at experimental applications. For developing the efficient method to identify the stage of sperm cells, the morphological characteristics of sperm cells were investigated by staining the cells with blue fluorescent dye Hoechst 33258, and a criterion for male germ cell classification was elicited from results of the previous investigation, then the efficiency of the criterion was verified by applying it to assort the germ cells recovered from male mice in age from 6 to 35 days. As morphological characteristics, spermatogonia significantly differed from spermatocytes in size, appearance and fluorescent patches of nucleus, and spermatids could also be distinguished from spermatozoa by making a difference in the volume and shape of nucleus and the shape and fluorescence of tail. Aforesaid criterion was applicable for classifying in vitro cultured sperm cells by verifying its efficiency and propriety for assorting the stages of testicular germ cells. However, the fluorescent staining showed that germ cells in mouse testis should be dramatically differentiated and developed at 21 days and 35 days of age, which were known as times of sexual puberty and maturity in male mice, respectively. In conclusion, the results indicated that this simple criterion for sperm cell classification using fluorescence staining with Hoechst 33258 may be highly efficient and reasonable for spermatogenesis study.

Tra2${\alpha}$ and hnRNP K might be Functional Partners of Rbm for Regulation of RNA Processes during Spermatogenesis

  • Lee, Jungmin;Kim, Euisu;Jang, Sung Key;Rhee, Kunsoo
    • Animal cells and systems
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    • v.8 no.1
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    • pp.65-70
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    • 2004
  • Rbm is a male infertility gene located in the AZFb region of the Y chromosome. Expression pattern of Rbm indicates that Rbm is critical for early phase of male germ cell development. It shares strong structural homology with hnRNP G, suggesting a function as an RNA processing factor. In order to gain a clue on the molecular mechanisms of Rbm on male germ cell development, we examined interactions of Rbm with selected proteins in yeast. The results revealed specific interactions between Rbm, hnRNP K and Tra2${\alpha}$. These results suggest that hnRNP K and Tra2${\alpha}$ may be functional partners of Rbm in male germ cells. We propose a model in which hnRNP K may playa role as a platform for Rbm and Tra2${\alpha}$.

Germ Cell Apoptosis in the Testis of Transgenic Pigs

  • Chung, Hak-Jae;Kim, Bong-Ki;Ko, Yeoung-Gyu;Woo, Jei-Hyun;Kim, Jeom-Soon;Jung, Jin-Kwan;Chang, Won-Kyong
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.233-233
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    • 2004
  • PURPOSE: Gene expression and apoptosis in testicular germ cells has been demonstrated in many transgenic animals. However, little is known about the transgenic pig and rates of apoptosis during spermatogenesis. METHODS : Morphological and biochemical features of apoptosis reported in other species were used to confirm that the TdT-mediated dUTP Nick end labeling (TUNEL) assay is an acceptable mothos for idendtification and quantification of apoptotic transgenic germ cells in histological tissue section from transgenic pig testis. (omitted)

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Germ Cell Differentiations during Spermatogenesis and Taxonomic Values of Mature Sperm Morphology of $Atrina$ ($Servatrina$) $pectinata$ (Bivalvia, Pteriomorphia, Pinnidae)

  • Kang, Hee-Woong;Chung, Ee-Yung;Kim, Jin-Hee;Chung, Jae-Seung;Lee, Ki-Young
    • Development and Reproduction
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    • v.16 no.1
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    • pp.19-29
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    • 2012
  • The ultrastructural characteristics of germ cell differentiations during spermatogenesis and mature sperm morphology in male $Atrina$ ($Servatrina$) $pectinata$ were evaluated via transmission electron microscopic observation. The accessory cells, which contained a large quantity of glycogen particles and lipid droplets in the cytoplasm, are assumed to be involved in nutrient supply for germ cell development. Morphologically, the sperm nucleus and acrosome of this species are ovoid and conical in shape, respectively. The acrosomal vesicle, which is formed by two kinds of electron-dense or lucent materials, appears from the base to the tip: a thick and slender elliptical line, which is composed of electron-dense opaque material, appears along the outer part (region) of the acrosomal vesicle from the base to the tip, whereas the inner part (region) of the acrosomal vesicle is composed of electron-lucent material in the acrosomal vesicle. Two special characteristics, which are found in the acrosomal vesicle of A. ($S$) $pectinata$ in Pinnidae (subclass Pteriomorphia), can be employed for phylogenetic and taxonomic analyses as a taxonomic key or a significant tool. The spermatozoa were approximately $45-50{\mu}m$ in length, including a sperm nucleus (about $1.43{\mu}m$ in length), an acrosome (about $0.51{\mu}m$ in length), and a tail flagellum (about $46-47{\mu}m$). The axoneme of the sperm tail evidences a 9+2 structure.

Screening of Chicken Genes Related to Germ Cell Development (닭에서 생식세포 발달에 관여하는 유전자 검색)

  • Lee, Jee-Young;Kim, Hee-Bal;Kim, Duk-Kyung;Song, Ki-Duk;Lim, Jeong-Mook;Han, Jae-Yong
    • 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.

Comparative Analysis for In Vitro Differentiation Potential of Induced Pluripotent Stem Cells, Embryonic Stem Cells, and Multipotent Spermatogonial Stem Cells into Germ-lineage Cells

  • Go, Young-Eun;Kim, Hyung-Joon;Jo, Jung-Hyun;Lee, Hyun-Ju;Do, Jeong-Tae;Ko, Jung-Jae;Lee, Dong-Ryul
    • Development and Reproduction
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    • v.15 no.1
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    • pp.41-52
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    • 2011
  • In the present study, embryoid bodies (EBs) obtained from induced pluripotent stem cells (iPSCs) were induced to differentiate into germ lineage cells by treatment with bone morphogenetic protein 4 (BMP4) and retinoic acid (RA). The results were compared to the results for embryonic stem cells (ESCs) and multipotent spermatogonial stem cells (mSSCs) and quantified using immunocytochemical analysis of germ cell-specific markers (integrin-${\alpha}6$, GFR-${\alpha}1$, CD90/Thy1), fluorescence activating cell sorting (FACS), and real time-RT-PCR. We show that the highest levels of germ cell marker-expressing cells were obtained from groups treated with 10 ng/$m{\ell}$ BMP4 or 0.01 ${\mu}M$ RA. In the BMP4-treated group, GFR-${\alpha}1$ and CD90/Thy-1 were highly expressed in the EBs of iPSCs and ESCs compared to EBs of mSSCs. The expression of Nanog was much lower in iPSCs compared to ESCs and mSSCs. In the RA treated group, the level of GFR-${\alpha}1$ and CD90/Thy-1 expression in the EBs of mSSCs Induced pluripotent stem cells, Mouse embryonic stem cells, Multipotent spermatogonial stem cells, Germ cell lineage, Differentiation potential. was much higher than the levels found in the EBs of iPSCs and similar to the levels found in the EBs of ESCs. FACS analysis using integrin-${\alpha}6$, GFR-${\alpha}1$, CD90/Thy1 and immunocytochemistry using GFR-${\alpha}1$ antibody showed similar gene expression results. Therefore our results show that iPSC has the potential to differentiate into germ cells and suggest that a protocol optimizing germ cell induction from iPSC should be developed because of their potential usefulness in clinical applications requiring patient-specific cells.

A Case of Primary Mediastinal Germ Cell Tumor Associated with Klinefelter's Syndrome (Klinefelter 증후군에 병발된 원발성 종격동 생식세포종 1예)

  • Kim, Yong-Jo;Kwun, Gyo-Seon;Lee, Young Wo;Kim, Kyung-Tae;Park, Yeon-Hee;Ryoo, Baek-Yeol;Kim, Tae You;Im, Young-Hyuck;Lee, Choon-Taek;Kang, Yoon-Koo;Cho, Kyung Ja;Lee, Jhin-Oh;Kang, Tae-Woong
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.6
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    • pp.1035-1041
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    • 1996
  • Klinefelter's syndrome is characterized by small testes, azoospermia, gynecomastia, and elevated levels of plasma gonadotropins in men with two or more X chromosomes. Previous investigators reponed that patients with Klinefelter's syndrome are predisposed to the development of a non-seminomatous germ cell tumor in the mediastinum. It is suggested that this linkage may be due to the hormonal imbalance in Klinefelter's syndrome and consequently, the formation of dysgenetic germ cell and/or abnomal migration of germ cell We report here a case of Klinefelter's syndrome in a 24-years-old man who was presented with anterior mediastinal mass. The clinical and laborarotory fmdings were compatible with Klinefelter's syndrome and he was found to have 47 XXY karyotype. Pathological findings for mediastinal mass revealed mixed germ cell tumor composed of mature cystic teraloma and endodermal sinus rumor. He was treated with cis platin containing chemotherapy and followed up in partial remission.

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Expression of Murine Asb-9 During Mouse Spermatogenesis

  • Lee, Man Ryul;Kim, Soo Kyoung;Kim, Jong Soo;Rhim, Si Youn;Kim, Kye-Seong
    • Molecules and Cells
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    • v.26 no.6
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    • pp.621-624
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    • 2008
  • We previously showed that Asb-4 and Asb-17 is uniquely expressed in developing male germ cells. A recent report showed that Asb-9 is specifically expressed in the kidney and testes; however, detailed expression patterns in developing germ cells have not been shown. Northern blot analysis in various tissues demonstrated that mAsb-9 was strongly expressed in the testes. Expression analysis by RT-PCR and Northern blot in developing mouse testes indicates that mAsb-9 is expressed from the fourth week after birth to adulthood, with the highest expression in round spermatids. Expression sites were further localized by in situ hybridization in the testes. Pachytene spermatocytes and spermatids expressed mAsb-9 but spermatogonia and generated spermatozoa did not. This study reveals that mAsb-9 could be a specific marker of active spermatogenesis and would be useful for studies of male germ cell development.

Sacrococcygeal Teratoma : A Tumor at the Center of Embryogenesis

  • Phi, Ji Hoon
    • Journal of Korean Neurosurgical Society
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    • v.64 no.3
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    • pp.406-413
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    • 2021
  • Sacrococcygeal teratoma (SCT) is an extragonadal germ cell tumor (GCT) that develops in the fetal and neonatal periods. SCT is a type I GCT in which only teratoma and yolk sac tumors arise from extragonadal sites. SCT is the most common type I GCT and is believed to originate through epigenetic reprogramming of early primordial germ cells migrating from the yolk sac to the gonadal ridges. Fetal SCT diagnosed in utero presents many obstetrical problems. For high-risk fetuses, fetal interventions (devascularization and debulking) are under development. Most patients with SCT are operated on after birth. Complete surgical resection is the key for tumor control, and the anatomical location of the tumor determines the surgical approaches. Incomplete resection and malignant histology are risk factors for recurrence. Approximately 10-15% of patients have a tumor recurrence, which is frequently of malignant histology. Long-term surveillance with monitoring of serum alpha fetoprotein and magnetic resonance imaging is required. Survivors of SCT may suffer anorectal, urological, and sexual sequelae later in their life, and comprehensive evaluation and care are required.