Knockdown of zebrafish Nanog increases primordial germ cells during early embryonic development

Wang, Huannan; Liu, Yanhua; Ye, Ding; Li, Jianzhen; Liu, Jiangdong; Deng, Fang

doi:10.1111/dgd.12279

Cited by 19 publications

(22 citation statements)

References 27 publications

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“…The maternal nanog and ndr1 transcripts were deposited in the embryo in agreement with the previous studies (Lee et al, 2013;Leichsenring et al, 2013;Liang & Rubinstein, 2003;Rebagliati, Toyama, Fricke, et al, 1998;H. Wang et al, 2016).…”

Section: Discussionsupporting

confidence: 91%

The effect of Activin pathway modulation on the expression of both pluripotency and differentiation markers during early zebrafish development compared with other vertebrates

et al. 2021

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Activin-like factors control many developmental processes, including pluripotency maintenance and differentiation. Although Activin-like factors' action in mesendoderm induction has been demonstrated in zebrafish, their involvement in preserving the stemness remains unknown. To investigate the role of maternal Activin-like factors, their effects were promoted or blocked using synthetic human Activin A or SB-431542 treatments respectively until the maternal to zygotic transition. To study the role of zygotic Activin-like factors, SB-431542 treatment was also applied after the maternal to zygotic transition. The effect of the pharmacological modulations of the Activin/Smad pathway was then studied on the mRNA expressions of the ndr1, ndr2, tbxta (no tail/ntl) as the differentiation index, mych, nanog, and oct4 (pou5f3) as the pluripotency markers of the zebrafish embryonic cells as well as sox17 as a definitive endoderm marker. Expression of the target genes was measured at the 16-cell, 256-cell, 1K-cell, oblong, dome, and shield stages using the real-time quantitative polymerase chain reaction (RT-qPCR). Activation of the maternal Activin signaling pathway led to an increase in zygotic expression of the tbxta, particularly marked at the oblong stage. In other words, promotion of the maternal Activin/Smad pathway induced differentiation by advancing the major peaks of ndr1 and nanog, thereby eliciting tbxta expression. Whereas suppression of the maternal or zygotic Activin/Smad pathway sustained the pluripotency by preventing the major peaks of ndr1 and nanog as well as tbxta encoding.

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Section: Discussionsupporting

confidence: 91%

The effect of Activin pathway modulation on the expression of both pluripotency and differentiation markers during early zebrafish development compared with other vertebrates

et al. 2021

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“…7d). As nanog mRNA levels are only robust before 6 hpf and decline dramatically afterwards 69 , such deep sequencing results at 10 and 24 hpf might not reflect situations when the embryos were undergoing the dorsoventral patterning. Notably, the transcripts of all ribosomal proteins were downregulated by 87% to 57% (mean = 68%) at 24 hpf (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…75 ), smad5 (refs. 76–78 ), and nanog 69 . Although detailed molecular mechanisms are still unclear, the reduced levels of these ventralization factors (Fig.…”

Section: Discussionmentioning

confidence: 99%

Regulation of zebrafish dorsoventral patterning by phase separation of RNA-binding protein Rbm14

et al. 2019

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RNA-binding proteins with intrinsically disordered regions (IDRs) such as Rbm14 can phase separate in vitro. To what extent the phase separation contributes to their physiological functions is however unclear. Here we show that zebrafish Rbm14 regulates embryonic dorsoventral patterning through phase separation. Zebrafish rbm14 morphants displayed dorsalized phenotypes associated with attenuated BMP signaling. Consistently, depletion of mammalian Rbm14 downregulated BMP regulators and effectors Nanog, Smad4/5, and Id1/2, whereas overexpression of the BMP-related proteins in the morphants significantly restored the developmental defects. Importantly, the IDR of zebrafish Rbm14 demixed into liquid droplets in vitro despite poor sequence conservation with its mammalian counterpart. While its phase separation mutants or IDR failed to rescue the morphants, its chimeric proteins containing an IDR from divergent phase separation proteins were effective. Rbm14 complexed with proteins involved in RNA metabolism and phase separated into cellular ribonucleoprotein compartments. Consistently, RNA deep sequencing analysis on the morphant embryos revealed increased alternative splicing events as well as large-scale transcriptomic downregulations. Our results suggest that Rbm14 functions in ribonucleoprotein compartments through phase separation to modulate multiple aspects of RNA metabolism. Furthermore, IDRs conserve in phase separation ability but not primary sequence and can be functionally interchangeable.

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“…Nanog mRNA and protein are present in the germline of the ovary and testes (Wang et al, 2016). Therefore, Nanog might be required during oogenesis and the earliest developmental stages.…”

Section: Cell Death At the End Of Gastrulation Is Increased In Nanog mentioning

confidence: 99%

Maternal Nanog is required for zebrafish embryo architecture and for cell viability during gastrulation

et al. 2018

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Nanog has been implicated in establishment of pluripotency in mammals and in zygotic genome activation in zebrafish. In this study, we characterize the development of MZ (maternal and zygotic null) mutant zebrafish embryos Without functional Nanog, epiboly is severely affected, embryo axes do not form and massive cell death starts at the end of gastrulation. We show that three independent defects in MZ mutants contribute to epiboly failure: yolk microtubule organization required for epiboly is abnormal, maternal mRNA fails to degrade owing to the absence of miR-430, and actin structure of the yolk syncytial layer does not form properly. We further demonstrate that the cell death in MZ embryos is cell-autonomous. Nanog is necessary for correct spatial expression of the ventral-specifying genes , and , and the neural transcription factor It is also required for the correctly timed activation of endoderm genes and for the degradation of maternal mRNA via miR-430 Our findings suggest that maternal Nanog coordinates several gene regulatory networks that shape the embryo during gastrulation.

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Knockdown of zebrafish Nanog increases primordial germ cells during early embryonic development

Cited by 19 publications

References 27 publications

The effect of Activin pathway modulation on the expression of both pluripotency and differentiation markers during early zebrafish development compared with other vertebrates

The effect of Activin pathway modulation on the expression of both pluripotency and differentiation markers during early zebrafish development compared with other vertebrates

Regulation of zebrafish dorsoventral patterning by phase separation of RNA-binding protein Rbm14

Maternal Nanog is required for zebrafish embryo architecture and for cell viability during gastrulation

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