Summary
It is possible to distinguish the morphological features of the spermatogonial nuclei and nucleoli and to further identify their distinct generations using an appropriate method to fix whole testes via vascular perfusion with glutaraldehyde, postfixation by immersion in reduced osmium, embedding in araldite, and staining of semithin tissue sections. A well-trained individual can distinguish each of the spermatogonial types in rodents (Aundiferentiated, A1, A2, A3, A4, In, and B) using this tissue preparation technique based on their morphological details and without correlation with the stages of the epithelium cycle or other parameters. The possibility of distinguishing each spermatogonial type by their morphological characteristics allows a more accurate evaluation of their kinetics during the spermatogenic cycle. Moreover, the understanding of spermatogonial behavior is a means to elucidate the functional control of the spermatogenesis, which consequently allows the determination of their effects on the fertility of humans and other animals.
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Acknowledgments
The standardizing of this method is in part because of the efforts of Lonnie D. Russell (in memoriam), who applied it with a unique ability to show us under HRLM for the first time various important parameters of male reproductive biology. We thank Fernanda RCL Almeida and Deborah Amaral for their suggestions on the manuscript. This method was standardized during the development of different projects that were partially supported by Brazilian (CAPES, FAPEMIG, CNPq) and U.S. (National Institutes of Health, Latin American Fellowship) financial foundations.
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Chiarini-Garcia, H., Meistrich, M.L. (2008). High-Resolution Light Microscopic Characterization of Spermatogonia. In: Hou, S.X., Singh, S.R. (eds) Germline Stem Cells. Methods in Molecular Biology™, vol 450. Humana Press. https://doi.org/10.1007/978-1-60327-214-8_6
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DOI: https://doi.org/10.1007/978-1-60327-214-8_6
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