Temporal and spatial localization of three germline‐specific proteins in medaka

Aoki, Yumiko; Nagao, Ippei; Saito, Daisuke; Ebe, Youko; Kinjo, Masataka; Tanaka, Minoru

doi:10.1002/dvdy.21448

Cited by 48 publications

(32 citation statements)

References 28 publications

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“…In mice, vasa is also localized in the cytoplasm of germ cells [13]. It is also known that other germ cell-specific proteins, such as nanos and trdr, are predominantly localized in the cytoplasm of germ cells [14,15], although these chicken homologue genes have not yet been investigated. Whereas vasa, nanos and trdr are conserved among many species, including drosophila, dead end orthologs have not been identified in invertebrates [1].…”

Section: Discussionmentioning

confidence: 99%

Chicken Dead End Homologue Protein is a Nucleoprotein of Germ Cells Including Primordial Germ Cells

Aramaki

Kubota

Soh

et al. 2009

Journal of Reproduction and Development

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Abstract. The dead end gene, coding an RNA binding protein, is predominantly expressed in the germ cells of vertebrates. Recently, we cloned chicken dead end homologue (CDH) and showed that expression of CDH mRNA is highly specific to primordial germ cells (PGCs) at early embryonic stages. To date, the subcelluler localization of Dead end protein in germ cell has been largely unknown due to lack of an antibody. Here, we raised a polyclonal antibody against chicken dead end homologue (CDH) to elucidate its subcellular localization in the germ cells. For comparative studies with CDH, a polyclonal antibody against chicken vasa homologue (CVH), a well-known germ cell marker, was also raised. Immunoblotting analysis for CDH protein showed a single band with a molecular size of approximately 60 kDa in the ovarian and testicular proteins. Immunofluorescence studies revealed that CDH protein was exclusively localized in the nuclei of primordial germ cells (PGCs) and germ cells at later stages, while CVH was localized in the cytoplasm. Interestingly, the germ cells distributed at the basal sides of seminiferous epithelia, such as spermatogonia, were strongly positive to CDH protein. The current study provides novel evidence that CDH is a nucleoprotein of germ cells, including PGCs.

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Section: Discussionmentioning

confidence: 99%

Chicken Dead End Homologue Protein is a Nucleoprotein of Germ Cells Including Primordial Germ Cells

Aramaki

Kubota

Soh

et al. 2009

Journal of Reproduction and Development

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“…A cDNA clone for Sdgc (clone name: olte54h09) was obtained from NBRP medaka (http://www.shigen.nig.ac.jp/medaka/). For immunohistochemistry, anti-OLVAS (medaka Vasa antibody, 1:100, rat) (Aoki et al, 2008) or anti-GFP (1:100, mouse; Clontech, 632381) was used as a primary antibody that was detected with Alexa 488-conjugated secondary antibodies (1:100, goat; Life Technologies, A11029).…”

Section: In Situ Hybridization and Immunohistochemistrymentioning

confidence: 99%

Analysis of a novel gene, Sdgc, reveals sex chromosome-dependent differences of medaka germ cells prior to gonad formation

et al. 2014

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In vertebrates that have been examined to date, the sexual identity of germ cells is determined by the sex of gonadal somatic cells. In the teleost fish medaka, a sex-determination gene on the Y chromosome, DMY/dmrt1bY, is expressed in gonadal somatic cells and regulates the sexual identity of germ cells. Here, we report a novel mechanism by which sex chromosomes cell-autonomously confer sexually different characters upon germ cells prior to gonad formation in a genetically sexdetermined species. We have identified a novel gene, Sdgc (sex chromosome-dependent differential expression in germ cells), whose transcripts are highly enriched in early XY germ cells. Chimeric analysis revealed that sexually different expression of Sdgc is controlled in a germ cell-autonomous manner by the number of Y chromosomes. Unexpectedly, DMY/dmrt1bY was expressed in germ cells prior to gonad formation, but knockdown and overexpression of DMY/dmrt1bY did not affect Sdgc expression. We also found that XX and XY germ cells isolated before the onset of DMY/dmrt1bY expression in gonadal somatic cells behaved differently in vitro and were affected by Sdgc. Sdgc maps close to the sex-determination locus, and recombination around the two loci appears to be repressed. Our results provide important insights into the acquisition and plasticity of sexual differences at the cellular level even prior to the developmental stage of sex determination.

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“…1, 3) followed a previously described method (Nakamura et al, 2006). The primary antibodies used in this study were anti-GFP antibody (1/100; Nacalai Tesque, Kyoto, Japan) and anti-tdrd1 serum (1/100; Aoki et al, 2008;Nakamura et al, 2008a). All images were taken with FV1000 confocal laser microscopy (Olympus, Tokyo, Japan).…”

Section: Ihc In Situ Hybridization and Histologymentioning

confidence: 99%

Two distinct types of theca cells in the medaka gonad: Germ cell‐dependent maintenance of cyp19a1‐expressing theca cells

Nakamura

Kurokawa

Asakawa

et al. 2009

Developmental Dynamics

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Aromatase is a steroidogenic enzyme catalyzing the production of estrogens and is important for the proper development and function of the reproductive system. The lineage of cyp19a1 (ovarian-type aromatase)-expressing cells in the developing gonad was analyzed using a transgenic medaka (Oryzias latipes) that recapitulates endogenous cyp19a1 expression with EGFP fluorescence. Our results show that cyp19a1-expressing cells arise in the ventral stromal cells of the developing female gonad, then expand anteriorly as the gonadal region extends anteriorly. These cells become located close to the developing follicles, and are distinguishable from the P450c17-I-expressing theca cells. In the adult ovary, the expression of P450c17-I and cyp19a1 are mutually exclusive in the outer theca-cell layer. Cyp19a1 expression in the granulosa cells is found only in the population of large follicles. These observations demonstrate two types of theca cells in the medaka ovary. We also show that the maintenance of cyp19a1-expressing cells depends on germ cells.

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Temporal and spatial localization of three germline‐specific proteins in medaka

Cited by 48 publications

References 28 publications

Chicken Dead End Homologue Protein is a Nucleoprotein of Germ Cells Including Primordial Germ Cells

Chicken Dead End Homologue Protein is a Nucleoprotein of Germ Cells Including Primordial Germ Cells

Analysis of a novel gene, Sdgc, reveals sex chromosome-dependent differences of medaka germ cells prior to gonad formation

Two distinct types of theca cells in the medaka gonad: Germ cell‐dependent maintenance of cyp19a1‐expressing theca cells

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