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Antitubulin immunofluorescence studies of spermatogenesis in the mouse

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Abstract

Immunofluorescence staining techniques, using antibodies against tubulin molecules, have been successfully used with somatic tissue culture cells. Their application to gametogenesis, however, has been more difficult, largely due to the presence of many different cell types in such a preparation. We have circumvented this problem by counterstaining with dilute hematoxylin, a biological stain that shows excellent cellular morphology without affecting the fluorescence of the antibody. We can then photograph the same field, using either ultraviolet light or conventional bright-field microscopy. Using this approach, we have examined the entire progression of murine spermatogenesis, from spermatogonial cells through mature spermatozoa. This technique has been especially valuable in the visualization of the manchette, and has allowed a reassessment of the staging of spermatid development. In the future, the antibody/hematoxylin double-staining approach will allow a more informative examination of the effects of tubulin-active mitotic poisons on mammalian germline cells.

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Authors and Affiliations

  1. Department of Genetics, The University of Texas, M.D. Anderson Hospital and Tumor Institute at Houston, 77030, Houston, TX, USA

    Lorraine M. Cherry & T. C. Hsu

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  1. Lorraine M. Cherry
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  2. T. C. Hsu
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Cherry, L.M., Hsu, T.C. Antitubulin immunofluorescence studies of spermatogenesis in the mouse. Chromosoma 90, 265–274 (1984). https://doi.org/10.1007/BF00287034

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  • DOI: https://doi.org/10.1007/BF00287034

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