Spermatogonial deubiquitinase USP9X is essential for proper spermatogenesis in mice

Kishi, Kasane; Uchida, Aya; Takase, Hinako M.; Suzuki, Hitomi; Kurohmaru, Masamichi; Tsunekawa, Naoki; Kanai‐Azuma, Masami; Wood, Stephen A.; Kanai, Yoshikatsu

doi:10.1530/rep-17-0184

Cited by 28 publications

(27 citation statements)

References 41 publications

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“…The testes were fixed in Bouin's or 4% paraformaldehyde solution, dehydrated, embedded in paraffin, and serially sectioned (4 µm in thickness) (Kishi et al, ; Uchida et al, ). The sections were used for immunostaining with anti‐phospho‐STAT3 [Tyr705] (p‐STAT3; 1:200 dilution; Cell Signaling) or anti‐STAT3 (1:400 dilution; Cell Signaling) monoclonal antibody at 4 °C for 12 hr.…”

Section: Methodsmentioning

confidence: 99%

Regionally distinct patterns of STAT3 phosphorylation in the seminiferous epithelia of mouse testes

Nagasawa

Imura-Kishi

Uchida

et al. 2018

Molecular Reproduction Devel

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In mouse testes, Sertoli cells support the continuous process of spermatogenesis, which is dependent on seminiferous epithelial cycles along the longitudinal axis of the seminiferous tubule. Sertoli cell function is modulated partly by local cytokines and/or growth factors derived from adjacent tissues such as blood vessels, macrophages, rete testis, etc. However, the spatial activation patterns by local signals in vivo remain unclear. In this study, we focused on Signal Transducers and Activators of Transcription (STAT) signaling in Sertoli cells, because STAT is a major crucial cytokine transducer for somatic cyst cell regulation in Drosophila testis niches. In mouse testes, STAT3 was ubiquitously expressed in Sertoli cells throughout the seminiferous tubules. Phosphorylated STAT3 (p-STAT3) was predominantly observed in the Sertoli cells within the valve-like structure adjacent to the rete testis (i.e., the Sertoli valve [SV]) in the terminal segment of the proximal seminiferous tubules. In the distal seminiferous tubules with active spermatogenesis, most Sertoli cells were negative for anti-p-STAT3 staining. Albeit rarely, a small patch of several p-STAT3-positive Sertoli cells was detected frequently in seminiferous epithelial cycle stages I-VI. Such p-STAT3-positive ratios in the convoluted seminiferous epithelia were significantly increased in germ cell-less testes than in the wild-type testes, but with considerably lower ratios than in the SV region. These findings imply that regionally distinct patterns of STAT3 phosphorylation in the Sertoli cells depend on either location or spermatogenic activity in normal healthy testes in vivo, highlighting a novel entry point to understanding STAT signaling in mammalian spermatogenesis.

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Section: Methodsmentioning

confidence: 99%

Regionally distinct patterns of STAT3 phosphorylation in the seminiferous epithelia of mouse testes

Nagasawa

Imura-Kishi

Uchida

et al. 2018

Molecular Reproduction Devel

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“…We provide evidence that enhanced mTORC1 activation following GILZ loss underlies SSC exhaustion plus increased progenitor proliferation, and we link aberrant ERK MAPK signalling to mTORC1 dysregulation. Additionally, we uncover a role for GILZ in expression of USP9X and ZMYM3, essential regulators of spermatogenesis (Hu et al, 2017;Kishi et al, 2017). USP9X can also regulate mTORC1, suggesting that GILZ controls mTORC1 through multiple mechanisms (Fig.…”

Section: Discussionmentioning

confidence: 76%

“…Knockout of Zmym3, which is primarily expressed in progenitors, results in meiotic arrest and spermatocyte apoptosis (Hu et al, 2017). USP9X is present throughout the spermatogonial pool and germline Usp9x deletion triggers spermatocyte depletion (Kishi et al, 2017). GILZ may therefore regulate spermatogenesis through ZMYM3 and USP9X.…”

Section: Discussionmentioning

confidence: 99%

“…GILZ may therefore regulate spermatogenesis through ZMYM3 and USP9X. However, spermatogonial populations are preserved in Zmym3 and Usp9x knockout models (Hu et al, 2017;Kishi et al, 2017), indicating that other effectors are relevant for GILZ function in SSCs. Interestingly, USP9X regulates pathways with known or potential roles in SSCs by deubiquitylating and stabilising protein components, e.g.…”

Section: Discussionmentioning

confidence: 99%

“…USP9X is a deubiquitylase that regulates cellular pathways by modulating protein stability (Bridges et al, 2017;Premarathne et al, 2017). Germline Usp9x deletion blocks spermatogenesis and is associated with spermatocyte apoptosis (Kishi et al, 2017). USP9X colocalised with GILZ in the cytosol of spermatogonia and spermatocytes, and was expressed throughout the spermatogonial hierarchy ( Fig.…”

Section: Identification Of Gilz Effector Genes In Spermatogoniamentioning

confidence: 99%

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GILZ-dependent modulation of mTORC1 regulates spermatogonial maintenance

Chan

Legrand

et al. 2018

Development

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Male fertility is dependent on spermatogonial stem cells (SSCs) that self-renew and produce differentiating germ cells. Growth factors produced within the testis are essential for SSC maintenance but intrinsic factors that dictate the SSC response to these stimuli are poorly characterised. Here, we have studied the role of GILZ, a TSC22D family protein and spermatogenesis regulator, in spermatogonial function and signalling. Although broadly expressed in the germline, GILZ was prominent in undifferentiated spermatogonia and deletion in adults resulted in exhaustion of the GFRα1 SSC-containing population and germline degeneration. GILZ loss was associated with mTORC1 activation, suggesting enhanced growth factor signalling. Expression of deubiquitylase USP9X, an mTORC1 modulator required for spermatogenesis, was disrupted in mutants. Treatment with an mTOR inhibitor rescued GFRα1 spermatogonial failure, indicating that GILZ-dependent mTORC1 inhibition is crucial for SSC maintenance. Analysis of cultured undifferentiated spermatogonia lacking GILZ confirmed aberrant activation of ERK MAPK upstream mTORC1 plus USP9X downregulation and interaction of GILZ with TSC22D proteins. Our data indicate an essential role for GILZ-TSC22D complexes in ensuring the appropriate response of undifferentiated spermatogonia to growth factors via distinct inputs to mTORC1.

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TGFBR2 is a novel substrate and indirect transcription target of deubiquitylase USP9X in granulosa cells

Yang

Wang

Pan

et al. 2022

Journal Cellular Physiology

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The ubiquitin‐specific peptidase 9 X‐linked (USP9X) is one of the highly conserved members belonging to the ubiquitin‐specific proteases (USPs) family, which has been reported to control substrates‐mediated biological functions through deubiquitinating and stabilizing substrates. Here, we have found that TGFBR2, the type II receptor of the transforming growth factor beta (TGF‐β) signaling pathway, is a novel substrate and indirect transcription target of deubiquitylase USP9X in granulosa cells (GCs). Mechanically, USP9X positively influences the expression of TGFBR2 at different levels through two independent ways: (i) directly targets and deubiquitinates TGFBR2, which maintains the protein stability of TGFBR2 through avoiding degradation mediated by ubiquitin‐proteasome system; (ii) indirectly maintains TGFBR2 messenger RNA (mRNA) expression via SMAD4/miR‐143 axis. Specifically, SMAD4, another substrate of USP9X, acts as a transcription factor and suppresses miR‐143 which inhibits the mRNA level of TGFBR2 by directly binding to its 3′‐untranslated region. Functionally, the maintenance of TGFBR2 by USP9X activates the TGF‐β signaling pathway, which further represses GC apoptosis. Our study highlights a functional micro‐regulatory network composed of deubiquitinase (USP9X), small noncoding RNA (miR‐143) and the TGF‐β signaling pathway, which plays a crucial role in the regulation of GC apoptosis and female fertility.

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Spermatogonial deubiquitinase USP9X is essential for proper spermatogenesis in mice

Cited by 28 publications

References 41 publications

Regionally distinct patterns of STAT3 phosphorylation in the seminiferous epithelia of mouse testes

Regionally distinct patterns of STAT3 phosphorylation in the seminiferous epithelia of mouse testes

GILZ-dependent modulation of mTORC1 regulates spermatogonial maintenance

TGFBR2 is a novel substrate and indirect transcription target of deubiquitylase USP9X in granulosa cells

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