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The expression patterns of aquaporin 9, vacuolar H+-ATPase, and cytokeratin 5 in the epididymis of the common vampire bat

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Abstract

Desmodus rotundus is a vampire bat species that inhabits Latin America. Some basic aspects of this species’ biology are still unknown, as the histophysiological characteristics of the male reproductive tract. Our study has focused on its epididymis, which is an important organ for performing a variety of functions, especially the sperm maturation and storage. The aim of this study was to identify principal, narrow, clear, and basal cells using cell-specific markers such as aquaporin 9 (AQP9), vacuolar H+-ATPase (V-ATPase), and cytokeratin 5 (KRT5). Principal cells were labeled by AQP9 from initial segment to cauda region in their stereocilia. They were shown with a columnar shape, whereas V-ATPase-rich cells were identified with a goblet-shaped body along the entire epididymis, including the initial segment, which were named as clear cells. Pencil-shaped V-ATPase-rich cells (narrow cells) were not detected in the initial segment of the bat epididymis, unlike in the rodent. Basal cells were labeled by KRT5 and were located at the basal portion of the epithelium forming a dense network. However, no basal cells with a luminal-reaching body extension were observed in the bat epididymis. In summary, epithelial cells were identified by their specific markers in the vampire bat epididymis. Principal and basal cells were labeled by AQP9 and KRT5, respectively. Narrow cells were not observed in the vampire bat epididymis, whereas clear cells were identified by V-ATPase labeling along the entire duct in a goblet-shaped body. In addition, no luminal-reaching basal cells were observed in the vampire bat epididymis.

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Acknowledgments

The authors thank Jerusa M. Oliveira, Bruno E.S. Melo, and Susana P. Ribeiro for their support during the collection of the bats. The study was carried out in the Program in Membrane Biology (PMB) Laboratory at Massachusetts General Hospital and supported by National Institutes of Health (NIH) grants HD040793 and DK097124 (to Sylvie Breton.); Mariana M. Castro has been granted a fellowship of Brazilian Federal Agency for Support and Evaluation of Graduate School (CAPES) and participated as a visiting graduate student in the PMB with scholarship from CAPES (Process Number—99999.011031/2013-01).

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    Authors and Affiliations

    1. Department of General Biology, Federal University of Viçosa, Viçosa, MG, 36570-000, Brazil

      Mariana M. Castro & Mariana Machado-Neves

    2. Center for Systems Biology, Program in Membrane Biology and Division of Nephrology, Department of Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA, 02114, USA

      Bongki Kim, Eric Hill & Sylvie Breton

    3. Department of Morphology, Federal University of Amazonas, Manaus, AM, 69077-000, Brazil

      Maria C. Q. Fialho

    4. Instituto Mineiro de Agropecuária, Minas Gerais, Juiz de Fora, MG, 36016-011, Brazil

      Luciano C. H. P. Puga

    5. Department of Animal Biology, Federal University of Viçosa, Viçosa, MG, 36570-000, Brazil

      Mariella B. Freitas

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    1. Mariana M. Castro
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    Mariana M. Castro and Bongki Kim have contributed equally to this work.

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    Supplementary Fig. 1

    Negative controls performed in Desmodus rotundus’ epididymis omitting all primary antibodies and using as secondary antibody Alexa 488-conjugated goat anti-rabbit IgG (AQP9, Cldn1, KRT5) and CY3-conjugated donkey anti-chicken (V-ATPase) and anti-rat IgG (ZO-1). Sperm and nuclei are labeled with DAPI (blue). Scales 50 µm (TIFF 11769 kb)

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    Castro, M.M., Kim, B., Hill, E. et al. The expression patterns of aquaporin 9, vacuolar H+-ATPase, and cytokeratin 5 in the epididymis of the common vampire bat. Histochem Cell Biol 147, 39–48 (2017). https://doi.org/10.1007/s00418-016-1477-9

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