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Immunohistochemical Analysis of Histone H3 Modifications in Germ Cells during Mouse Spermatogenesis

By Ning Song, Jie Liu, Shucai An, Tomoya Nishino, Yoshitaka Hishikawa and Takehiko Koji

Abstract

Histone modification has been implicated in the regulation of mammalian spermatogenesis. However, the association of differently modified histone H3 with a specific stage of germ cells during spermatogenesis is not fully understood. In this study, we examined the localization of variously modified histone H3 in paraffin-embedded sections of adult mouse testis immunohistochemically, focusing on acetylation at lysine 9 (H3K9ac), lysine 18 (H3K18ac), and lysine 23 (H3K23ac); tri-methylation at lysine 4 (H3K4me3) and lysine 27 (H3K27me3); and phosphorylation at serine 10 (H3S10phos). As a result, we found that there was a significant fluctuation in the modifications; in spermatogonia, the stainings for H3K9ac, H3K18ac, and H3K23ac were strong while that for H3K4me3 was weak. In spermatocytes, the stainings for H3K9ac, H3K18ac, H3K23ac, and H3K4me3 were reduced in the preleptotene to pachytene stage, but in diplotene stage the stainings for H3K18ac, H3K23ac, and H3K4me3 seemed to become intense again. The staining for H3K27me3 was nearly constant throughout these stages. In the ensuing spermiogenesis, a dramatic acetylation and methylation of histone H3 was found in the early elongated spermatids and then almost all signals disappeared in the late elongated spermatids, in parallel with the replacement from histones to protamines. In addition, we confirmed that the staining of histone H3S10phos was exclusively associated with mitotic and meiotic cell division. Based upon the above results, we indicated that the modification pattern of histone H3 is subject to dynamic change and specific to a certain stage of germ cell differentiation during mouse spermatogenesis

Topics: Regular Article
Publisher: Japan Society of Histochemistry and Cytochemistry
OAI identifier: oai:pubmedcentral.nih.gov:3168764
Provided by: PubMed Central

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Citations

  1. (2005). A histone code in meiosis: the histone kinase, NHK-1, is required for proper chromosomal architecture in Drosophila oocytes.Song et al. 190 Genes Dev.
  2. (2002). Active genes are tri-methylated at K4 of histone H3.
  3. (1992). Antibodies to defined histone epitopes reveal variations in chromatin conformation and underacetylation of centric heterochromatin in human metaphase chromosomes.
  4. (2005). Chromatin remodelling and epigenetic features of germ cells.
  5. (2004). Distinct localization of histone H3 acetylation and H3-K4 methylation to the transcription start sites in the human genome.
  6. (2004). Dynamic histone modifications mark sex chromosome inactivation and reactivation during mammalian spermatogenesis.
  7. (2008). Epigenetic mechanisms of gene regulation during mammalian spermatogenesis.
  8. (2007). Epigenetics: a landscape takes shape.
  9. (1987). Factors affecting nucleosome disassembly by protamines in vitro. Histone hyperacetylation and chromatin structure, time dependence, and the size of the sperm nuclear proteins.
  10. (2005). Genomic maps and comparative analysis of histone modifications in human and mouse.
  11. (2000). Histone acetylation and an epigenetic code.
  12. (2005). Histone H3 serine 10 phosphorylation by Aurora B causes HP1 dissociation from heterochromatin.
  13. (2003). Histone lysine methylation: a signature for chromatin function.
  14. (2009). Histone methylation sets the stage for meiotic DNA breaks.
  15. (2009). Human papillomavirus infection and its possible correlation with p63 expression
  16. (2009). Improvement of in situ PCR by optimization of PCR cycle number and proteinase k concentration: localization of x chromosome-linked phosphoglycerate kinase-1 gene in mouse reproductive organs.
  17. (2008). In situ detection of methylated DNA by histo endonuclease-linked detection of methylated DNA sites: a new principle of analysis of DNA methylation.
  18. (2005). Induction of cell death in rat small intestine by ischemia reperfusion: differential roles of Fas/Fas ligand and Bcl-2/Bax systems depending upon cell types.
  19. (2002). Involvement of Bax redistribution in the induction of germ cell apoptosis in neonatal mouse testes.
  20. (2010). Jmjd1a demethylase-regulated histone modification is essential for cAMP-response element modulator-regulated gene expression and spermatogenesis.
  21. (2008). Lineagespecific polycomb targets and de novo DNA methylation define restriction and potential of neuronal progenitors.
  22. (2001). Loss of the Suv39h histone methyltransferases impairs mammalian heterochromatin and genome stability.
  23. (1986). Marked differences in the ability of distinct protamines to disassemble nucleosomal core particles in vitro.
  24. (1997). Mitosis-specific phosphorylation of histone H3 initiates primarily within pericentromeric heterochromatin during G2 and spreads in an ordered fashion coincident with mitotic chromosome condensation.
  25. (2006). Mouse polycomb proteins bind differentially to methylated histone H3 and RNA and are enriched in facultative heterochromatin.
  26. (2006). Pluripotency of spermatogonial stem cells from adult mouse testis.
  27. (2000). Semi-quantitative non-radioactive in situ hybridization and its clinical application.
  28. (2000). The language of covalent histone modifications.
  29. (1962). The process of spermatogenesis in mammals.
  30. (2004). The role of histones in chromatin remodelling during mammalian spermiogenesis.
  31. (2002). Unique chromatin remodeling and transcriptional regulation in spermatogenesis.
  32. (2010). VAMP2 marks quiescent satellite cells and myotubes, but not activated myoblasts.