Abstract
Structural and immunohistochemical methods have been extremely useful for the characterization of the XY body (the structure formed by the XY pair during meiotic prophase) in Man and in other mammals. These methods are widely used at the present time for the detection of abnormalities leading to human infertility. The basic ultrastructural methods are spreading of pachytene spermatocytes, thin-sectioning techniques with or without 3-D reconstructions, and the monitoring of all specimens with semi-thin sections. Immunofluorescent techniques also use spreading of meiotic cells for the analysis of the XY body, and they can be combined with the fluorescence in situ hybridization (FISH) technique, in the so-called immuno-FISH. Epitope retrieval techniques are also used.
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References
Solari AJ (1974) The behaviour of the XY pair in mammals. Int Rev Cytol 38:273–317
Solari AJ (1994) Sex chromosomes and sex determination in vertebrates. CRC Press, Boca Raton, FL
Franco MJ, Sciurano RB, Solari AJ (2007) Protein immunolocalization supports the presence of identical mechanisms of XY body formation in eutherians and marsupials. Chromosome Res 15:815–824
Sciurano RB, Rahn MI, Rossi L et al (2012) Synapsis, recombination, and chromatin remodeling in the XY body of armadillos. Chromosome Res 20:293–302
Monesi V (1965) Differential rate of ribonucleic acid synthesis in the autosomes and sex chromosomes during male meiosis in the mouse. Chromosoma 17:11–21
Handel MA (2004) The XY body: a specialized meiotic chromatin domain. Exp Cell Res 296:57–63
Ichijima Y, Sin HS, Namekawa SH (2012) Sex chromosome inactivation in germ cells: emerging roles of DNA damage response pathways. Cell Mol Life Sci 69:2559–2572
Coco R, Rahn MI, Estanga PG et al (2004) A constitutional complex chromosome rearrangement involving meiotic arrest in an azoospermic male: case report. Hum Reprod 19:2784–2790
Sciurano RB, Rahn MI, Pigozzi MI et al (2006) An azoospermic man with a double-strand DNA break-processing deficiency in the spermatocyte nuclei: case report. Hum Reprod 21:1194–1203
Burgoyne PS, Mahadevaiah SK, Turner JM (2009) The consequences of asynapsis for mammalian meiosis. Nat Rev Genet 10:207–216
Sciurano RB, Rahn MI, Rey-Valzacchi G et al (2012) The role of asynapsis in human spermatocyte failure. Int J Androl 35:541–549
Anderson LK, Reeves A, Webb LM et al (1999) Distribution of crossing over on mouse synaptonemal complexes using immunofluorescent localization of MLH1 protein. Genetics 151:1569–1579
Reynolds ES (1963) The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol 17:208–212
Sciurano RB, Luna Hisano CV, Rahn MI et al (2009) Focal spermatogenesis originates in euploid germ cells in classical Klinefelter patients. Hum Reprod 24:2353–2360
Evans EP, Breckon G, Ford CE (1964) An air-drying method for meiotic preparations from mammalian testes. Cytogenetics 15:289–294
Sudman PD (1989) Cryogenic preservation of mammalian testicular material for synaptonemal complex analysis. Cytogenet Cell Genet 52:88–89
Howell WM, Black DA (1980) Controlled silver staining of nucleolus organizer regions with a protective colloidal developer: a 1 step method. Experientia 36:1014–1015
Solari AJ (1998) Structural analysis of meiotic chromosomes and synaptonemal complexes in higher vertebrates. Methods Cell Biol 53:236–254
Acknowledgments
We thank M.Sc. I.M. Rahn for the useful comments on the manuscript and for experimental help. Thanks to Mrs C. Deparci for the able technical help. We acknowledge Prof. R. Ponzio for the histopathological diagnosis of the biopsies, and R. Coco, Ph.D.; G. Rey Valzacchi, M.D.; and S. Brugo Olmedo, M.D., for the clinical and surgical procedures. We also thank the generous gift of antibodies from P. J. Moens, B. Spyropoulus (York University, Toronto, Ontario, Canada), and M. Alsheimer (University of Würzburg, Würzburg, Germany). This work was supported by UBACYT 20020100100030 (AJS) and FONCYT PICT 2010-2718 (RBS).
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Sciurano, R.B., Solari, A.J. (2014). Ultrastructural and Immunofluorescent Methods for the Study of the XY Body as a Biomarker. In: Stockert, J., Espada, J., Blázquez-Castro, A. (eds) Functional Analysis of DNA and Chromatin. Methods in Molecular Biology, vol 1094. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-706-8_11
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DOI: https://doi.org/10.1007/978-1-62703-706-8_11
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