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Proteome-wide identification of palmitoylated proteins in mouse testis

  • Male Reproduction: Original Article
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Abstract

The reversible lipid modification, S-palmitoylation, plays regulatory roles in various physiological processes, e.g., neuronal plasticity and organs development; however, the roles of palmitoylation engaged in testis have yet remained unexplored. Here, we used combined approaches of palm-proteomics, informatics and quantitative PCR to systematically analyze the expression of key enzymes related to protein palmitoylation and identify proteome-wide palmitoylated proteins during the processes of spermatogenesis. Specifically, different timepoints were chosen to collect samples to cover the initiation of meiosis (postnatal, P12), the appearance of the first batch of sperm (P36) and fully fertile status (P60) in mouse. Interestingly, our results showed that only a few enzymes related to protein palmitoylation are highly expressed at later stages (from P36 to P60), rather than in the earlier phase of testis development (P12). To focus on the molecular event of spermatogenesis, we examined the palm-proteomics of testes in P36 and P60 mouse. In total, we identified 4,883 palmitoylated proteins, among which 3,310 proteins match the published palmitoyl-proteome datasets and 1,573 proteins were firstly identified as palmitoylated proteins in this study. Informatics analysis suggested that palmitoylation is involved in events of protein transport, metabolic process, protein folding and cell adhesion, etc. Importantly, further analysis revealed that several networks of palmitoylated proteins are closely associated with sperm morphology and motility. Together, our study laid a solid ground for understanding the roles of protein palmitoylation in spermatogenesis for future studies.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 31870773 to Z.J.Z. and No. 31770824 to E.Y.K.), Key Technologies Research and Development Program of Henan Province (212102311072) and Ph.D. Research Program of Xinxiang Medical University (505318) to W.S.G.

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Authors and Affiliations

  1. The Second Affiliated Hospital of Xinxiang Medical University, Xinxiang Medical University, Xinxiang, 453000, China

    Jun Gao & Eryan Kong

  2. Institute of Psychiatry and Neuroscience, Xinxiang Medical University, Xinxiang, 453000, China

    Jun Gao, Wenchao Li, Zhongjian Zhang & Eryan Kong

  3. Department of Epidemiology and Health Statistics, School of Public Health, Xinxiang Medical University, Xinxiang, 453000, China

    Wenshan Gao

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  1. Jun Gao
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  2. Wenchao Li
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  3. Zhongjian Zhang
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Contributions

J.G. was involved in data acquisition and analysis, validation and writing. W.C.L. was involved in methodology and data acquisition; W.S.G. was involved in data curation and software analysis. Z.J.Z. was involved in resources, funding and project administration. E.Y.K. was involved in conceptualization, funding acquisition, and writing—review & editing. All authors approved the final version.

Corresponding authors

Correspondence to Wenshan Gao or Eryan Kong.

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Ethics Approval

All animal procedures were performed according to guidelines approved by the committee on animal care at Xinxiang medical university. The approval number of the animal experiment is XXLL-20210312.

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All the authors listed have approved the manuscript that is enclosed.

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The authors declare no competing interests.

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Gao, J., Li, W., Zhang, Z. et al. Proteome-wide identification of palmitoylated proteins in mouse testis. Reprod. Sci. 29, 2299–2309 (2022). https://doi.org/10.1007/s43032-022-00919-w

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  • DOI: https://doi.org/10.1007/s43032-022-00919-w

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