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Scaffold protein Lin7 family in membrane skeletal protein complex in mouse seminiferous tubules

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Abstract

The membrane skeletal complex, protein 4.1G–membrane palmitoylated protein 6 (MPP6), is localized in spermatogonia and early spermatocytes of mouse seminiferous tubules. In this study, we investigated the Lin7 family of scaffolding proteins, which interact with MPP6. By immunohistochemistry, Lin7a and Lin7c were localized in germ cells, and Lin7c had especially strong staining in spermatogonia and early spermatocytes, characterized by staging of seminiferous tubules. By immunoelectron microscopy, Lin7 localization appeared under cell membranes in germ cells. The Lin7 staining pattern in seminiferous tubules was partially similar to that of 4.1G, cell adhesion molecule 1 (CADM1), and melanoma cell adhesion molecule (MCAM). Lin7-positive cells included type A spermatogonia, as revealed by double staining for Lin28a. Lin7 staining became weaker in MPP6-deficient mice by immunohistochemistry and western blotting, indicating that MPP6 transports and maintains Lin7 in germ cells. The histology of seminiferous tubules was unchanged in MPP6-deficient mice compared to that of wild-type mice. In cultured spermatogonial stem cells maintained with glial cell line-derived neurotropic factor (GDNF), Lin7 was clearly expressed and immunolocalized along cell membranes, especially at cell–cell junctions. Thus, Lin7 protein is expressed in germ cells, and Lin7, particularly Lin7c, is a useful marker for early spermatogenesis.

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Funding

This work was partially supported by a grant from the Japan Society for the Promotion of Science (KAKENHI 16K08463) to N. Terada.

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Authors and Affiliations

  1. Health Science Division, Department of Medical Sciences, Shinshu University Graduate School of Medicine, Science and Technology, 3-1-1 Asahi, Matsumoto City, Nagano, 390-8621, Japan

    Akio Kamijo, Yurika Saitoh & Nobuo Terada

  2. Center for Medical Education, Teikyo University of Science, Adachi-ku, Tokyo, Japan

    Yurika Saitoh

  3. Division of Cellular and Molecular Biology, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo, Japan

    Takeharu Sakamoto

  4. Laboratory of Cell and Molecular Biology, Department of Animal Science, School of Veterinary Medicine, Kitasato University, Towada, Aomori, Japan

    Hiroshi Kubota

  5. Laboratory of Molecular Neuroscience and Neurology, School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, Hachioji City, Tokyo, Japan

    Junji Yamauchi

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  1. Akio Kamijo
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  2. Yurika Saitoh
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  3. Takeharu Sakamoto
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  4. Hiroshi Kubota
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  5. Junji Yamauchi
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  6. Nobuo Terada
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Correspondence to Nobuo Terada.

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Kamijo, A., Saitoh, Y., Sakamoto, T. et al. Scaffold protein Lin7 family in membrane skeletal protein complex in mouse seminiferous tubules. Histochem Cell Biol 152, 333–343 (2019). https://doi.org/10.1007/s00418-019-01807-2

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