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Methods Mol Biol. 2018;1832:293-307. doi: 10.1007/978-1-4939-8663-7_16.

Characterization of Post-Meiotic Male Germ Cell Genome Organizational States.

Author information

1
Large Scale Biology Laboratory, Biosciences and Biotechnologies Institute of Grenoble, INSERM U1038, CEA, Université Grenoble Alpes, Grenoble, France.
2
Institut Albert Bonniot, CNRS UMR 5309, INSERM U1209, Université Grenoble Alpes, Grenoble, France.
3
Institut Albert Bonniot, CNRS UMR 5309, INSERM U1209, Université Grenoble Alpes, Grenoble, France. saadi.khochbin@univ-grenoble-alpes.fr.

Abstract

Dramatic and unique genome reorganizations accompany the differentiation of haploid male germ cells, characterized by a gradual loss of the vast majority of histones leading to a final tight compaction of the genome by protamines. Despite being essential for procreation and the life cycle, the mechanisms driving the transformation of nucleosomes into nucleoprotamines remain poorly understood. To address this issue, our laboratory has developed a number of specific approaches, ranging from the purification of spermatogenic cells at specific stages, the analysis of chromatin transitional states, the functional characterization of histone variants, histone-replacing proteins and their chaperones. This chapter will detail all related relevant techniques with a particular emphasis on methods allowing the functional studies of histone variants and the genome organizational states associated with the studied histones in spermatogenic cells undergoing histone-to-protamine exchange.

KEYWORDS:

Chaperones; Chromatin; Germline; Histone; Histone variants; Nucleosome; Protamines; Sperm; Spermatogenesis; Transition proteins

PMID:
30073534
DOI:
10.1007/978-1-4939-8663-7_16
[Indexed for MEDLINE]

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