UPF2, a nonsense-mediated mRNA decay factor, is required for prepubertal Sertoli cell development and male fertility by ensuring fidelity of the transcriptome

Bao, Jianqiang; Tang, Chong; Yuan, Shuiqiao; Porse, Bo T.; Yan, Wei

doi:10.1242/dev.115642

Cited by 34 publications

(38 citation statements)

References 65 publications

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“…Interestingly, one study showed the transcription of Amh in Leydig cells, questioning its selectivity to Sertoli cells (Ao, Erickson, & Stalvey, 1993). However, the purity of Leydig cell primary culture samples was not extensively evaluated in that study, and the results obtained could most likely be due to contamination by Sertoli cells (Bao et al, 2015;Garcia et al, 2014;Sridharan, Brehm, Bergmann, & Cooke, 2007). Another line of evidence for Sertoli cell-specificity of AMH comes from the extensive use of the Amh-cre deleter mouse line (Lécureuil, Fontaine, Crepieux, & Guillou, 2002) by investigators to specifically express or delete genes in Sertoli cells; no off-target effects are noted in Leydig cells, demonstrating the absence of AMH expression in Leydig cells (Garcia et al, 2014;Hurtado et al, 2018;Sridharan et al, 2007).…”

Section: Markers Used To Evaluate the Purity Of Rodent Sertoli Cellmentioning

confidence: 96%

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Characterization of rodent Sertoli cell primary cultures

Zomer

Reddi

2020

Molecular Reproduction Devel

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Sertoli cells play a vital role in spermatogenesis by offering physical and nutritional support to the differentiating male germ cells. They form the blood–testis barrier and secrete growth factors essential for germ cell differentiation. Sertoli cell primary cultures are critical for understanding the regulation of spermatogenesis; however, obtaining pure cultures has been a challenge. Rodent Sertoli cell isolation protocols do not rule out contamination by the interstitial or connective tissue cells. Sertoli cell‐specific markers could be helpful, but there is no consensus. Vimentin, the most commonly used marker, is not specific for Sertoli cells since its expression has been reported in peritubular myoid cells, mesenchymal stem cells, fibroblasts, macrophages, and endothelial cells, which contaminate Sertoli cell preparations. Markers based on transcription and growth factors also have limitations. Thus, the impediment to obtaining pure Sertoli cell cultures pertains to both the method of isolation and marker usage. The aim of this review is to discuss improvements to current methods of rodent Sertoli cell primary cultures, assess the properties of prepubertal versus mature Sertoli cell cultures, and propose steps to improve cellular characterization. Potential benefits of using contemporary approaches, including lineage tracing, specific cell ablation, and RNA‐seq for obtaining Sertoli‐specific transcript markers are discussed. Evaluating the specificity and applicability of these markers at the protein level to characterize Sertoli cells in culture would be critical. This review is expected to positively impact future work using primary cultures of rodent Sertoli cells.

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Section: Markers Used To Evaluate the Purity Of Rodent Sertoli Cellmentioning

confidence: 96%

“…The anti-Mullerian hormone (AMH), also known as Mullerian inhibiting substance, is an early Sertoli cell cytoplasmic protein with expression starting at embryonic day E12.5 that gradually decreases during Sertoli cell maturation(Bao, Tang, Yuan, Porse, & Yan, 2015; …”

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confidence: 99%

Characterization of rodent Sertoli cell primary cultures

Zomer

Reddi

2020

Molecular Reproduction Devel

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“…3D). In general, 1 1 transcripts containing a PTC are subjected to rapid degradation by NMD (29)(30)(31).…”

Section: Inefficiency Of Nmd Substantially Contributes To Accumulatiomentioning

confidence: 99%

Identification of germ cell-specificMgavariant mRNA that promotes meiotic entry via impediment of a non-canonical PRC1

Kitamura

Uranishi

Hirasaki

et al. 2020

Preprint

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Transition from mitosis to meiosis in cell division is a fundamental process of gametogenesis. This transition is thought to be largely controlled by the exchange of relative dominance between positive and negative regulation by the retinoic acid/Stra8 signal cascade and a non-canonical PRC1 (PRC1.6), respectively. We have previously demonstrated that germ cells have transcriptionally and/or post-translationally reduced levels of MAX, a component of PRC1.6, immediately prior to meiotic onset, leading to alleviation of the negative effect of PRC1.6 against meiotic onset. Here, we found that germ cells produced Mga variant mRNA bearing a premature termination codon (PTC) during meiosis as an additional mechanism to impede the function of PRC1.6. Our data indicated that spermatocytes and/or round spermatids produced an anomalous MGA protein lacking the bHLHZ domain from the variant mRNA and therefore functioned as a dominant negative regulator of PRC1.6 by exquisitely using their inefficient background of PTC-mediated nonsense-mediated mRNA decay. Thus, our data indicate that meiotic onset of male germ cells is controlled in a multi-layered manner in which both MAX and MGA, which constitute the core of PRC1.6 by their interaction, are at least used as targets to deteriorate the integrity of the complex to ensure initiation of meiosis.Significance StatementPRC1.6, a non-canonical PRC1, functions as a strong blocker of meiotic onset. Therefore, germ cells need to alleviate the function of the complex as a prerequisite for meiotic onset. The MGA/MAX heterodimer not only constitutes a core of PRC1.6, but also confers direct DNA-binding activity to the complex. We have previously demonstrated that germ cells reduce Max amounts prior to meiotic onset to inactivate PRC1.6. In this study, we explored the possibility of an additional molecular mechanism that promotes meiotic onset via impediment of PRC1.6 functions as a safeguard system. Here, we demonstrate that meiotic germ cells specifically generate variant Mga mRNA by alternative splicing, which leads to production of a dominant negative regulator of PRC1.6.

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“…Recently, an essential role for UPF2 has also been associated with male fertility. Conditional ablation of UPF2 in mouse embryonic Sertoli cells caused severe testicular atrophy, leading to sterility in adulthood …”

Section: Biological Functions Of Nmdmentioning

confidence: 99%

“…Conditional ablation of UPF2 in mouse embryonic Sertoli cells caused severe testicular atrophy, leading to sterility in adulthood. 146 SMG6 knockout mice also show early embryonic lethality, but using a conditional knockout strategy, SMG6 −/− mouse embryonic stem cells (ESCs) could be generated that proliferated normally and were morphologically indistinguishable from control ESCs. 147 Interestingly, such cultured SMG6 knockout ESCs were unable to undergo differentiation and retained high expression of pluripotency markers, and analysis of chimeric mice showed that the SMG6 −/− ESCs failed to differentiate into all three germ layers.…”

Section: Nmd Factors Are Required For Mammalian Developmentmentioning

confidence: 99%

Nonsense‐mediated mRNA decay: novel mechanistic insights and biological impact

2016

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Nonsense-mediated mRNA decay (NMD) was originally coined to define a quality control mechanism that targets mRNAs with truncated open reading frames due to the presence of a premature termination codon. Meanwhile, it became clear that NMD has a much broader impact on gene expression and additional biological functions beyond quality control are continuously being discovered. We review here the current views regarding the molecular mechanisms of NMD, according to which NMD ensues on mRNAs that fail to terminate translation properly, and point out the gaps in our understanding. We further summarize the recent literature on an ever-rising spectrum of biological processes in which NMD appears to be involved, including homeostatic control of gene expression, development and differentiation, as well as viral defense.

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UPF2, a nonsense-mediated mRNA decay factor, is required for prepubertal Sertoli cell development and male fertility by ensuring fidelity of the transcriptome

Cited by 34 publications

References 65 publications

Characterization of rodent Sertoli cell primary cultures

Characterization of rodent Sertoli cell primary cultures

Identification of germ cell-specificMgavariant mRNA that promotes meiotic entry via impediment of a non-canonical PRC1

Nonsense‐mediated mRNA decay: novel mechanistic insights and biological impact

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