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miRNA Profiling of Major Testicular Germ Cells Identifies Stage-Specific Regulators of Spermatogenesis

  • Male Reproduction: Original Article
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Abstract

Spermatogenesis is tightly controlled at transcriptional, post-transcriptional, and epigenetic levels by various regulators, including miRNAs. This study deals with the identification of miRNAs critical to the three important stages of germ cell development (spermatocytes, round spermatids, and mature sperm) during spermatogenesis. We used high-throughput transcriptome sequencing to identify the differentially expressed miRNAs in the pachytene spermatocytes, round spermatids, and mature sperm of rat. We identified 1843 miRNAs that were differentially expressed across the three stages of germ cell development. These miRNAs were further categorized into three classes according to their pattern of expression during spermatogenesis: class 1 — miRNAs found exclusively in one stage and absent in the other two stages; class 2 — miRNAs found in any two stages but absent in the third stage; class 3 — miRNAs expressed in all the three stages. Six hundred forty-six miRNAs were found to be specific to one developmental stage, 443 miRNAs were found to be common across any two stages, and 754 miRNAs were common to all the three stages. Target prediction for ten most abundant miRNAs specific to each category identified miRNA regulators of mitosis, meiosis, and cell differentiation. The expression of each miRNA is specific to a particular developmental stage, which is required to maintain a significant repertoire of target mRNAs in the respective stage. Thus, this study provided valuable data that can be used in the future to identify the miRNAs involved in spermatogenic arrest at a particular stage of the germ cell development.

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Data Availability

All representative data are provided within the manuscript and also in the supporting information. Sequencing data can be provided upon request.

Code Availability

Not applicable.

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Acknowledgements

Shruti Sethi and Poonam Mehta would like to thank the University Grants Commission for graduate fellowship (191620102885/CSIR-UGC NET DEC.2019) and (460/CSIR-UGC NET DEC.2017), respectively.

Funding

The authors are thankful to the Council of Scientific and Industrial Research (CSIR) for funding this study under MLP2026.

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Authors and Affiliations

  1. CSIR-Central Drug Research Institute, Lucknow, India

    Shruti Sethi, Poonam Mehta, Aastha Pandey, Gopal Gupta & Singh Rajender

  2. Academy of Scientific and Innovative Research [AcSIR], Ghaziabad, India

    Shruti Sethi, Poonam Mehta, Aastha Pandey, Gopal Gupta & Singh Rajender

Authors
  1. Shruti Sethi
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  2. Poonam Mehta
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  3. Aastha Pandey
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  4. Gopal Gupta
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  5. Singh Rajender
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Contributions

SR and GG conceived the idea. SS, PM, and AP have performed the experimental procedures. SS, PM, and RS have analysed the data. SS, PM, RS, and GG wrote the manuscript.

Corresponding author

Correspondence to Singh Rajender.

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Ethics Approval

The study was approved by the institutional ethics committee of CSIR-CDRI (IAEC/2014/49/Renew03(135/16)) and the study was performed in accordance with the ethical standards as mentioned in 1964 Declaration of Helsinki and its later amendments.

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The authors declare no competing interests.

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Sethi, S., Mehta, P., Pandey, A. et al. miRNA Profiling of Major Testicular Germ Cells Identifies Stage-Specific Regulators of Spermatogenesis. Reprod. Sci. 29, 3477–3493 (2022). https://doi.org/10.1007/s43032-022-01005-x

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  • DOI: https://doi.org/10.1007/s43032-022-01005-x

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