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Expression of insulin-like factor 3 hormone-receptor system in the reproductive organs of male goats

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Abstract

Relaxin-like factor (RLF), generally known as insulin-like factor 3 (INSL3), is essential for testis descent during fetal development. However, its role in adult males is not fully understood. We investigate the function of INSL3 in male Saanen goats by identifying cell types expressing its receptor, relaxin/insulin-like family peptide receptor (RXFP)2 and by characterizing the developmental expression pattern of INSL3 and RXFP2 and the binding of INSL3 to target cells in the male reproductive system. A highly specific RXFP2 antibody that co-localizes with an anti-FLAG antibody in HEK-293 cells recognizes RXFP2-transcript-expressing cells in the testis. INSL3 and RXFP2 mRNA expression is upregulated in the testis, starting from puberty. INSL3 mRNA and protein expression has been detected in Leydig cells, whereas RXFP2 mRNA and protein localize to Leydig cells, to meiotic and post-meiotic germ cells and to the epithelium and smooth muscle of the cauda epididymis and vas deferens. INSL3 binds to all of these tissues and cell types, with the exception of Leydig cells, in a hormone-specific and saturable manner. These results provide evidence for a functional intra- and extra-testicular INSL3 ligand-receptor system in adult male goats.

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Acknowledgments

The authors thank Mr. Y. Hattori (Keyence, Osaka, Japan) for the use of the BZ-9000 All-in-One fluorescence microscope and Dynamic cell count BZ-HIC software.

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Authors and Affiliations

  1. Animal Reproduction and Physiology, Faculty of Agriculture, Shizuoka University, Shizuoka, 422-8529, Japan

    Ali Mohammed Pitia, Itaru Minagawa, Naoto Uera & Tetsuya Kohsaka

  2. Division of Animal Resource Production, United Graduate School of Agricultural Science, Gifu University, Gifu, 501-1193, Japan

    Ali Mohammed Pitia, Itaru Minagawa & Tetsuya Kohsaka

  3. Faculty of Agriculture, Shinshu University, Minami-Minowa, Nagano, 399-4598, Japan

    Koh-Ichi Hamano

  4. Livestock Breeding Division, National Livestock Breeding Center, Nagano Station, Saku, 385-007, Japan

    Yasushi Sugawara & Yoshio Nagura

  5. Animal Science High-Tech Research Center, Kitasato University School of Veterinary Medicine, Towada, 034-8628, Japan

    Yoshihisa Hasegawa

  6. Veterinary Pathology, Kitasato University School of Veterinary Medicine, Towada, 034-8628, Japan

    Toshifumi Oyamada

  7. Animal Reproduction, Kitasato University School of Veterinary Medicine, Towada, 034-8628, Japan

    Hiroshi Sasada

  8. Animal Reproduction and Physiology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka, 422-8529, Japan

    Tetsuya Kohsaka

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  1. Ali Mohammed Pitia
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  2. Itaru Minagawa
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  3. Naoto Uera
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  4. Koh-Ichi Hamano
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  5. Yasushi Sugawara
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  6. Yoshio Nagura
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  7. Yoshihisa Hasegawa
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  8. Toshifumi Oyamada
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  9. Hiroshi Sasada
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  10. Tetsuya Kohsaka
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Correspondence to Tetsuya Kohsaka.

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This work was supported by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (nos. 24580408 and 15K07691 to T. Kohsaka).

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Pitia, A.M., Minagawa, I., Uera, N. et al. Expression of insulin-like factor 3 hormone-receptor system in the reproductive organs of male goats. Cell Tissue Res 362, 407–420 (2015). https://doi.org/10.1007/s00441-015-2206-8

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  • DOI: https://doi.org/10.1007/s00441-015-2206-8

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