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mTORC1/rpS6 and mTORC2/PKC regulate spermatogenesis through Arp3-mediated actin microfilament organization in Eriocheir sinensis

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Abstract

Mammalian target of rapamycin (mTOR) is a crucial signaling protein regulating a range of cellular events. Numerous studies have reported that the mTOR pathway is related to spermatogenesis in mammals. However, its functions and underlying mechanisms in crustaceans remain largely unknown. mTOR exists as two multimeric functional complexes termed mTOR complex 1 (mTORC1) and mTORC2. Herein, we first cloned ribosomal protein S6 (rpS6, a downstream molecule of mTORC1) and protein kinase C (PKC, a downstream effector of mTORC2) from the testis of Eriocheir sinensis. The dynamic localization of rpS6 and PKC suggested that both proteins may be essential for spermatogenesis. rpS6/PKC knockdown and Torin1 treatment led to defects in spermatogenesis, including germ cell loss, retention of mature sperm and empty lumen formation. In addition, the integrity of the testis barrier (similar to the blood-testis barrier in mammals) was disrupted in the rpS6/PKC knockdown and Torin1 treatment groups, accompanied by changing in expression and distribution of junction proteins. Further study demonstrated that these findings may result from the disorganization of filamentous actin (F-actin) networks, which were mediated by the expression of actin-related protein 3 (Arp3) rather than epidermal growth factor receptor pathway substrate 8 (Eps8). In summary, our study illustrated that mTORC1/rpS6 and mTORC2/PKC regulated spermatogenesis via Arp3-mediated actin microfilament organization in E. sinensis.

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Acknowledgements

The authors want to show their appreciation to all members of the Sperm Laboratory in Zhejiang University for their support, encouragement and assistant.

Funding

This project was supported by the National Natural Science Foundation of China (No.32072954 and 32270555).

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Authors and Affiliations

  1. The Sperm Laboratory, College of Life Sciences, Zhejiang University, Hangzhou, 310058, China

    Zhen-Fang Li, Hong-Yu Qi, Jia-Ming Wang, Zhan Zhao & Wan-Xi Yang

  2. The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, 310003, China

    Fu-Qing Tan

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  1. Zhen-Fang Li
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  2. Hong-Yu Qi
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  3. Jia-Ming Wang
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  4. Zhan Zhao
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Correspondence to Wan-Xi Yang.

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Li, ZF., Qi, HY., Wang, JM. et al. mTORC1/rpS6 and mTORC2/PKC regulate spermatogenesis through Arp3-mediated actin microfilament organization in Eriocheir sinensis. Cell Tissue Res 393, 559–575 (2023). https://doi.org/10.1007/s00441-023-03795-1

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