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Inactivation of testis-specific gene C4orf46 is dispensable for spermatogenesis and fertility in mouse

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Abstract

Several genes have been reported to be involved in spermatogenesis but their functional importance in male fertility is yet needed to be elucidated. Therefore, in current research, we focused to explore the in vivo role of evolutionary conserved and testis-specifically expressed, C4orf46, gene in male mouse fertility and spermatogenesis. The expression profile of C4orf46 is specific to testes and expressed in testes from 7 days of postpartum to onward. Thus, we generated the C4orf46 knockout mice by utilizing CRISPR/Cas9 genome editing technology and examined gene function in spermatogenesis and fertility. Surprisingly, C4orf46 knockout mice were completely fertile, displayed normal testes morphology, however, higher sperm contents were observed in knockout mice compared to wild type (WT) littermates. Subsequently, intact testis histology and architecture of seminiferous tubules were observed in C4orf46 knockout and WT mice. Similarly, sperm morphology and swimming velocity of C4orf46 knockout mice were comparable with the WT littermates. Furthermore, all type of germ cells ranging from spermatogonia to mature spermatozoa were observed in the testes and epididymis sections of C4orf46 knockout mice suggesting that disruption of C4orf46 did not impact spermatogenesis. Moreover, meiotic prophase I progression was normal, and each type of cell population was comparable between knockout and WT mice. Overall, finding from this research indicates that C4orf46 is not an essential gene for fertility in mice. This study will help researchers to avoid the repetition and duplication of efforts, and to explore the genes that are indispensable for spermatogenesis and male fertility.

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Funding

This work was supported by the National Key Research and Developmental Program of China (2018YFC1003403 and 2016YFC1000600), the National Natural Science Foundation of China (31890780 and 31630050), the Strategic Priority Research Program of the Chinese Academy of Sciences (XDB19000000) and the Fundamental Research Funds for the Central Universities (YD2070002006).

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Authors and Affiliations

  1. The First Affiliated Hospital of USTC, National Laboratory for Physical Sciences At Microscale, School of Life Sciences, University of Science and Technology of China, Hefei, 230027, China

    Basit Shah, Ranjha Khan, Wasim Shah, Sobia Dil & Qinghua Shi

  2. Department of Biological Sciences, International Islamic University, Sector H10, Islamabad, 44000, Pakistan

    Ayesha Aftab

  3. Department of Biotechnology and Genetic Engineering, Hazara University, Dhodial, Pakistan

    Manan Khan

Authors
  1. Basit Shah
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  2. Ranjha Khan
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  3. Wasim Shah
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  4. Ayesha Aftab
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  5. Manan Khan
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  6. Sobia Dil
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  7. Qinghua Shi
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Correspondence to Ranjha Khan or Qinghua Shi.

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335_2021_9879_MOESM1_ESM.tif

Figure S1 PAS staining (A) Representative images of PAS stained testicular sections from 10-week-old C4orf46+/+ and C4orf46−/− mice. (B) Higher magnification image of the rectangular area outlined with white boxes in A. Scale bars, 50 um (TIF 6942 kb)

335_2021_9879_MOESM2_ESM.tif

Figure S2 Sperm morphology and motility analyses (A) Sperm morphology of 10-week-old C4orf46+/+ and C4orf46−/− mice. Scale bars, 100 μm. (B) Percentage of average rate of motile sperm in C4orf46+/+ and C4orf46−/− mice. (C) Percentage of progressive motile sperm from C4orf46+/+ and C4orf46−/− mice (TIF 1443 kb)

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Shah, B., Khan, R., Shah, W. et al. Inactivation of testis-specific gene C4orf46 is dispensable for spermatogenesis and fertility in mouse. Mamm Genome 32, 364–370 (2021). https://doi.org/10.1007/s00335-021-09879-z

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  • DOI: https://doi.org/10.1007/s00335-021-09879-z

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