Mitf is a transcriptional activator of medaka germ genes in culture

Zhao, Haobin; Li, Mingyou; Purwanti, Yovita Ida; Liu, Rong; Chen, Tiansheng; Li, Zhendong; Hóng, Ní; Guan, Guijun; Yin, Ao; Xiao, Ling; Ge, Ruowen; Song, Jianxing; Hong, Yunhan

doi:10.1016/j.biochi.2011.11.007

Cited by 16 publications

(17 citation statements)

References 43 publications

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“…Several medaka germ genes have been identified including vasa [40], boule and dazl [4], and dnd [41]. A transcription factor for germ genes has recently been identified in medaka [42]. Gene knockdown by morpholino has proven a powerful tool in medaka for analyzing PGCs development.…”

Section: Introductionmentioning

confidence: 99%

Medaka piwi is Essential for Primordial Germ Cell Migration

Li¹,

Hóng²,

Gui

et al. 2012

CMM

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Piwi controls the number of primordial germ cells (PGCs) via protecting maternal mRNA from decay and adult germ stem cell division in Drosophila. In mouse and zebrafish, piwi controls maintenance and differentiation of adult germ stem cell during gametogenesis. Whether piwi plays a role in PGC development of vertebrates remains unsolved. We addressed this issue by using medaka (Oryzias latipes) as a vertebrate model. Molecular cloning, sequence comparison and analyses of genomic organization and chromosome synteny led to the identification in this fish of a single piwi gene, called Opiwi. By RT-PCR analyses and in situ hybridization, the Opiwi transcript is maternally supplied and becomes restricted to PGCs and the central nervous system (CNS). Opiwi knockdown did not prevent PGC formation even in the absence of any somatic structures but did significantly reduce the number of PGCs in vivo and in vitro and affect the distribution of PGCs in developing embryos. Surprisingly, depletion of zygotic Opiwi severely and specifically affected PGC migration. We conclude that Opiwi is required not only for determining the PGC number but also for controlling PGC migration. Our results demonstrate that piwi plays a generally conserved role in germ cell development from Drosophila to vertebrate and a specific role in PGC migration.

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Section: Introductionmentioning

confidence: 99%

Medaka piwi is Essential for Primordial Germ Cell Migration

Li¹,

Hóng²,

Gui

et al. 2012

CMM

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“…We use the fish medaka (Oryzias latipes) as a model to study stem cells [37][38][39][40] germ cells [40][41][42] and sexual development [43,44]. We have recently identified Mitf as a first transcriptional activator of germ genes [6]. Here we show that RA suppresses Mitf-activated germ gene expression in stem cell cultures and exhausts the spermatogonial stem cell pool in organ culture.…”

Section: Introductionmentioning

confidence: 94%

“…Six luciferase (Luc) reporter plasmids and the effector plasmid pXmitf used in this study have been described [6,48]. pXmitf expresses the Xiphophorus mitfm [49].…”

Section: Reporter and Effector Plasmidsmentioning

confidence: 99%

“…For reporter assays, cells were transfected in 96-well plates as described [6]. In some experiments, cells were treated with all-trans RA (Sigma).…”

Section: Cell Culture and Luciferase Reporter Assaymentioning

confidence: 99%

“…However, how these germ genes are regulated for expression at the transcriptional level has remained essentially unexplored. One exception is the recent identification of microphthalmiaassociated transcription factor (Mitf) as a transcription activator of germ gene expression in medaka [6]. Mitf is a basic helix-loop-helix/ leucine zipper protein and acts as a master regulator of melanocyte development [7,8].…”

Section: Introductionmentioning

confidence: 99%

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Retinoic Acid Regulates Germ Gene Transcription In Vitro and Spermatogenesis in Testicular Organ Culture

Hóng¹,

Li²

2012

Biol Syst Open Access

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The adult testis is the male reproductive organ that produces sex hormones for sexual behaviours and sperm in a well-controlled process called spermatogenesis. Spermatogenesis involves self-renewal and differentiation of the male germ stem cells spermatogonia, culminating in the production of sperm for germline transmission. Retinoic acid (RA) is essential for fate decision and meiosis entry of mammalian germ cells. The microphthalmia-associated transcription factor (Mitf) is a newly identified transcriptional activator of germ genes in the fish medaka (Oryzias latipes). Here we report an essential role of RA on Mitf-activated germ gene expression and spermatogenesis. Medaka germ gene promoters DAZ, DND and VAS were found to contain RA-responsive elements and to exhibit little Mitf-activated transcriptional activity in response to RA treatment in both medaka ES cells and male germ stem cells. A procedure for testicular organ culture was established, which allows for test-tube spermatogenesis over a 21-day period. By the ordered testicular architecture, cellular morphology and VAS-driven GFP expression, different spermatogenic stages were identifiable on testicular cross-sections. In organ culture, RA treatment generated testes completely free of spermatogonia, and reduced VAS-driven GFP expression in spermatogonia but enhanced its expression in meiotic and post-meiotic germ cells. Therefore, RA represses Mitf-activated germ gene expression in stem cell cultures and differentially regulates germ gene expression depending upon the stages of spermatogenesis, and represses spermatogonial stem cell maintenance in cultured testes of medaka as a lower vertebrate model. These results demonstrate that RA plays a highly conserved role in germ cell development from fish to mammals.

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