Autonomous Regulation of Sex-Specific Developmental Programming in Mouse Fetal Germ Cells1

Iwahashi, Kazuhiro; Yoshioka, Hirotaka; Low, Eleanor W.; McCarrey, John R.; Yanagimachi, Ryuzo; Yamazaki, Yukiko

doi:10.1095/biolreprod.107.062851

Cited by 14 publications

(23 citation statements)

References 51 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Primordial germ cell isolation and culture XY and XX PGCs were isolated from the gonads for in vitro culture as previously described [8,28,29]. Briefly, male and female gonads obtained from (CD-1 X OG2) F 1 fetuses at 11.5 and 12.5 dpc were dissected in Hepes-DMEM supplemented with 15% FBS and incubated in 0.2% Collagenase (Calbiochem, San Diego, CA) and Accumax (Innovative Cell Technologies, San Diego) for 10 min at 37…”

Section: Xy Genotypingmentioning

confidence: 99%

“…The primer set for Nanos2 (Mm02525720 s1) was obtained from Applied Biosystems (Waltham, MA). The results were normalized to levels detected for β-actin gene expression and the levels of transcripts were presented as relative expression to control groups which were arbitrarily set as 1.00 as previously shown [8,28,29].…”

Section: Xy Genotypingmentioning

confidence: 99%

“…In this study, we have used our PGC culture system to investigate the direct effect of FGF9 on determination of XY PGC fate [8,28,29]. We first examined dose-dependent effects of FGF9 on XY PGC differentiation.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Dose-dependent functions of fibroblast growth factor 9 regulate the fate of murine XY primordial germ cells†

Ulu

Kim

Yokoyama

et al. 2016

Biology of Reproduction

Self Cite

View full text Add to dashboard Cite

Male differentiation of primordial germ cells (PGCs) is initiated by the inhibition of entry into meiosis and exposure to male-inducing factor(s), which are regulated by somatic elements of the developing gonad. Fibroblast growth factor 9 (FGF9) produced by pre-Sertoli cells is essential for male gonadal differentiation and also contributes to survival and male differentiation of XY PGCs. However, it is not clear how FGF9 regulates PGC fate. Using a PGC culture system, we identified dose-dependent, fate-determining functions of FGF9 in XY PGCs. Treatment with low levels of FGF9 (0.2 ng/ml) increased expression of male-specific Dnmt3L and Nanos2 in XY PGCs. Conversely, treatment with high levels of FGF9 (25 ng/ml) suppressed male-specific gene expression and stimulated proliferation of XY PGCs. Western blotting showed that low FGF9 treatment enhanced p38 MAPK (mitogen-activated protein kinase) phosphorylation in the same cells. In contrast, high FGF9 treatment significantly stimulated the ERK (extracellular signal-regulated kinase)1/2 signaling pathway in XY PGCs. We investigated the relationship between the ERK1/2 signaling pathway stimulated by high FGF9 and regulation of PGC proliferation. An ERK1/2 inhibitor (U0126) suppressed the PGC proliferation that would otherwise be stimulated by high FGF9 treatment, and increased Nanos2 expression in XY PGCs. Conversely, a p38 MAPK inhibitor (SB202190) significantly suppressed Nanos2 expression that would otherwise be stimulated by low FGF9 in XY PGCs. Taken together, our results suggest that stage-specific expression of FGF9 in XY gonads regulates the balance between proliferation and differentiation of XY PGCs in a dose-dependent manner. Summary SentenceFGF9 directly regulates different fates of XY PGCs in a dose-dependent manner: low FGF9 treatment promotes p38 signaling pathway to induce PGC male differentiation, whereas high FGF9 treatment enhances ERK1/2 signaling pathway to proliferate PGCs. 122

show abstract

Section: Xy Genotypingmentioning

confidence: 99%

Section: Xy Genotypingmentioning

confidence: 99%

See 1 more Smart Citation

Dose-dependent functions of fibroblast growth factor 9 regulate the fate of murine XY primordial germ cells†

Ulu

Kim

Yokoyama

et al. 2016

Biology of Reproduction

Self Cite

View full text Add to dashboard Cite

show abstract

“…Spermatogonia remain at the G0 stage of the cell cycle until after birth and can subsequently initiate meiosis several days later [14]. During the G0 stage, male germ cells initiate de novo methylation of imprinted loci and establish male-specific methylation imprints at around birth [15][16][17].…”

Section: Introductionmentioning

confidence: 99%

Male Differentiation of Germ Cells Induced by Embryonic Age-Specific Sertoli Cells in Mice1

Ohta

Yamamoto

Lin

et al. 2012

Biology of Reproduction

Self Cite

View full text Add to dashboard Cite

Retinoic acid (RA) is a meiosis-inducing factor. Primordial germ cells (PGCs) in the developing ovary are exposed to RA, resulting in entry into meiosis. In contrast, PGCs in the developing testis enter mitotic arrest to differentiate into prospermatogonia. Sertoli cells express CYP26B1, an RA-metabolizing enzyme, providing a simple explanation for why XY PGCs do not initiate meios/is. However, regulation of entry into mitotic arrest is likely more complex. To investigate the mechanisms that regulate male germ cell differentiation, we cultured XX and XY germ cells at 11.5 and 12.5 days postcoitus (dpc) with an RA receptor inhibitor. Expression of Stra8, a meiosis initiation gene, was suppressed in all groups. However, expression of Dnmt3l, a male-specific gene, during embryogenesis was elevated but only in 12.5-dpc XY germ cells. This suggests that inhibiting RA signaling is not sufficient for male germ cell differentiation but that the male gonadal environment also contributes to this pathway. To define the influence of Sertoli cells on male germ cell differentiation, Sertoli cells at 12.5, 15.5, and 18.5 dpc were aggregated with 11.5 dpc PGCs, respectively. After culture, PGCs aggregated with 12.5 dpc Sertoli cells increased Nanos2 and Dnmt3l expression. Furthermore, these PGCs established male-specific methylation imprints of the H19 differentially methylated domains. In contrast, PGCs aggregated with Sertoli cells at late embryonic ages did not commit to the male pathway. These findings suggest that male germ cell differentiation is induced both by inhibition of RA signaling and by molecule(s) production by embryonic age-specific Sertoli cells.

show abstract

“…In addition, some miRNAs function in a fail-safe mechanism to silence mRNAs that are unwanted in specific cell types Cohen et al 2006). It seems likely that miR-29b may function in such a mechanism to regulate methylation of the genome, given that the amount of Dnmt3a mRNA in PGCs is similar in male and female mouse embryos at E13.5 but is greater in male than in female PGCs at E15.5 and E17.5 (Iwahashi et al 2007).…”

Section: Discussionmentioning

confidence: 99%

Potential role of miR-29b in modulation ofDnmt3aandDnmt3bexpression in primordial germ cells of female mouse embryos

Takada¹,

Berezikov²,

Choi³

et al. 2009

RNA

View full text Add to dashboard Cite

MicroRNAs (miRNAs) are a recently discovered class of small noncoding RNAs and are implicated in an increasing number of biological processes. To examine whether miRNAs might contribute to sexual differentiation, we performed expression profiling of miRNAs in mouse embryonic gonads with the use of a highly sensitive cloning method, mRAP. Our profiling data revealed substantial differences in the miRNA repertoire between male and female gonads at embryonic (E) day 13.5 (E13.5), suggesting that such differentially expressed miRNAs may function in sexual differentiation. Female-specific miRNAs included miR-29b, which also has been known to be expressed in a similar sex-dependent manner in the gonads of chicken embryos, suggestive of a conserved role in gonadogenesis. Transcripts of the human genes for the de novo methyltransferases DNMT3A and DNMT3B have been identified as targets of miR-29b, and we found that mouse miR-29b also negatively regulates Dnmt3a and Dnmt3b expression in luciferase reporter assays. We also found that miR-29b is expressed in mouse primordial germ cells (PGCs) at E13.5 and that its expression is up-regulated in a female-specific manner between E13.5 and E17.5, when male-specific de novo methylation of the PGC genome is known to occur. Our data thus suggest that miR-29b may play an important role in female gonadal development by targeting Dnmt3a and Dnmt3b and thereby modulating methylation of genomic DNA in PGCs.

show abstract

Autonomous Regulation of Sex-Specific Developmental Programming in Mouse Fetal Germ Cells1

Cited by 14 publications

References 51 publications

Dose-dependent functions of fibroblast growth factor 9 regulate the fate of murine XY primordial germ cells†

Dose-dependent functions of fibroblast growth factor 9 regulate the fate of murine XY primordial germ cells†

Male Differentiation of Germ Cells Induced by Embryonic Age-Specific Sertoli Cells in Mice1

Potential role of miR-29b in modulation ofDnmt3aandDnmt3bexpression in primordial germ cells of female mouse embryos

Contact Info

Product

Resources

About