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Heterochromatin and histone modifications in the germline-restricted chromosome of the zebra finch undergoing elimination during spermatogenesis

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Abstract

In the zebra finch (Taeniopygia guttata) a germline-restricted chromosome (GRC) is regularly present in males and females. While the GRC is euchromatic in oocytes, in spermatocytes this chromosome is cytologically seen as entirely heterochromatic and presumably inactive. At the end of male meiosis, the GRC is eliminated from the nucleus. By immunofluorescence on microspreads, we investigated HP1 proteins and histone modifications throughout male meiotic prophase, as well as in young spermatid stages after the GRC elimination. We found that in prophase spermatocytes the GRC chromatin differs from that of the regular chromosome complement. The GRC is highly enriched in HP1β and exhibits high levels of di- and tri-methylated histone H3 at lysine 9 and tri- and di-methylated histone H4 at lysine 20. The GRC does not exhibit neither detectable levels of di- and tri-methylated histone H3 at lysine 4 nor acetylated histone H4 at lysine 5 and 8. The results prove the heterochromatic organization of the GRC in male germline and strongly suggest its transcriptional inactive state during male prophase. Following elimination, in young spermatids the GRC lacks HP1β signals but maintains high levels of methylated histone H3 at lysine 9 and methylated histone H4 at lysine 20. The release of HP1 from the GRC with respect to its elimination is discussed.

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Acknowledgments

This work was supported with grant PIP 124 from the National Research Council (CONICET) and UBACyT M429 from the University of Buenos Aires to MIP and with grant BFU2008-02947-C02-02/BMC to CG from M.E.C of Spain.

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Authors and Affiliations

  1. Departamento de Proliferación Celular y Desarrollo, Centro de Investigaciones Biológicas, CSIC, 28040, Madrid, Spain

    Clara Goday

  2. Instituto de Investigaciones en Reproducción, Facultad de Medicina, UBA, Buenos Aires, Argentina

    María Inés Pigozzi

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  1. Clara Goday
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  2. María Inés Pigozzi
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Correspondence to María Inés Pigozzi.

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Communicated by P. Cohen

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Suppl Fig. 1

Double immunofluorescence labeling with γH2AX (red) and SMC3 (green) antibodies in male germ cells counterstained with DAPI (blue). (A-A’’) Spermatocyte at leptotene showing chromatin-wide staining with γH2AX. The chromatin of the GRC (arrow) is more brightly stained with DAPI and show weak labeling with γH2AX. (B-B´´) Late zygotene spermatocyte showing the persistence of γH2AX foci on unsynapsed axis (arrowheads). The GRC chromatin is devoid of labeling. C-C’’. The eliminated GRC (arrow) is strongly labeled with γH2AX. Bar indicates 10 µm (JPEG 87 kb)

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Goday, C., Pigozzi, M.I. Heterochromatin and histone modifications in the germline-restricted chromosome of the zebra finch undergoing elimination during spermatogenesis. Chromosoma 119, 325–336 (2010). https://doi.org/10.1007/s00412-010-0260-2

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