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Cellular and subcellular localization of endogenous phospholipase D6 in seminiferous tubules of mouse testes

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Abstract

Phospholipase D6 (PLD6) plays pivotal roles in mitochondrial dynamics and spermatogenesis, but the cellular and subcellular localization of endogenous PLD6 in testis germ cells is poorly defined. We examined the distribution and subcellular localization of PLD6 in mouse testes using validated specific anti-PLD6 antibodies. Ectopically expressed PLD6 protein was detected in the mitochondria of PLD6-transfected cells, but endogenous PLD6 expression in mouse testes was localized to the perinuclear region of pachytene spermatocytes, and more prominently, to the round (Golgi and cap phases) and elongating spermatids (acrosomal phase); these results suggest that PLD6 is localized to the Golgi apparatus. The distribution of PLD6 in the round spermatids partially overlapped with that of the cis-Golgi marker GM130, indicating that the PLD6 expression corresponded to the GM130-positive subdomains of the Golgi apparatus. Correlative light and electron microscopy revealed that PLD6 expression in developing spermatids was localized almost exclusively to several flattened cisternae, and these structures might correspond to the medial Golgi subcompartment; neither the trans-Golgi networks nor the developing acrosomal system expressed PLD6. Further, we observed that PLD6 interacted with tesmin, a testis-specific transcript necessary for successful spermatogenesis in mouse testes. To our knowledge, these results provide the first evidence of PLD6 as a Golgi-localized protein of pachytene spermatocytes and developing spermatids and suggest that its subcompartment-specific distribution within the Golgi apparatus may be related to the specific functions of this organelle during spermatogenesis.

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Funding

This research was funded by grants from the National Research Foundation of Korea (NRF) (grant numbers: NRF-2018R1A2B3002179 and NRF-2020R1A2B5B01001442).

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Author notes

  1. Mun-Yong Lee and Do Sik Min contributed equally to this study

Authors and Affiliations

  1. Department of Anatomy, Catholic Neuroscience Institute, College of Medicine, The Catholic University of Korea, Seoul, 06591, Republic of Korea

    Tae-Ryong Riew, Soojin Kim, Xuyan Jin & Mun-Yong Lee

  2. Department of Biomedicine and Health Sciences, College of Medicine, The Catholic University of Korea, Seoul, 06591, Republic of Korea

    Xuyan Jin & Mun-Yong Lee

  3. Integrative Research Support Center, Laboratory of Electron Microscope, College of Medicine, The Catholic University of Korea, Seoul, 06591, Republic of Korea

    Hong Lim Kim

  4. College of Pharmacy, Yonsei University, Incheon, 21983, Republic of Korea

    Won Chan Hwang, Minju Kang & Do Sik Min

  5. Department of Molecular Biology, College of Natural Science, Pusan National University, Busan, 46241, Republic of Korea

    Won Chan Hwang & Eun Sun Yang

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  1. Tae-Ryong Riew
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Correspondence to Mun-Yong Lee or Do Sik Min.

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All procedures and provisions for animal care were conducted in accordance with the Laboratory Animals Welfare Act, the Guide for the Care and Use of Laboratory Animals, and the Guidelines and Policies for Rodent Survival Surgery provided by the Institutional Animal Care and Use Committee (IACUC) at the College of Medicine of The Catholic University of Korea (Approval number: CUMS-2020–0041-01). All experimental protocols were approved by the IACUC. The IACUC and the Department of Laboratory Animals (DOLA) at the Catholic University of Korea, Songeui Campus, were accredited for the Korea Excellence Animal Laboratory Facility by the Korea Food and Drug Administration in 2017 and acquired full Association for the Assessment and Accreditation of Laboratory Animal Care (AAALAC) International accreditation in 2018.

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Riew, TR., Kim, S., Jin, X. et al. Cellular and subcellular localization of endogenous phospholipase D6 in seminiferous tubules of mouse testes. Cell Tissue Res 385, 191–205 (2021). https://doi.org/10.1007/s00441-021-03442-7

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  • DOI: https://doi.org/10.1007/s00441-021-03442-7

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