2-hydroxyglutarate controls centromere and heterochromatin conformation and function in the male germline

Abstract

2-hydroxyglutarate (2HG) is recognized as an epigenetic regulator in cancer and in a few physiological states. Of all organs, the testis harbors the highest levels of 2HG, yet it's putative functions in germ cell biology are unknown. Here we show that 2HG is generated in specific stages of the mouse germ cell lineage by the testis specific lactate dehydrogenase C (LDHC). LDHC is expressed in pachytene, diplotene and diakinesis (PDD) cells and unexpectedly enters nuclei where it localizes along chromosomes and centromeres. LDHC-generated L-2HG controls centromere compaction and pericentromeric heterochromatin organization through multiple effects including clustering of chromocenters, centromere and chromocenter condensation and expression of satellite RNAs. The involvement of L-2HG in the above functions was shown both in isolated PDD cells and in vivo and is specific to the L but not D enantiomer. Our findings reveal that 2HG can rapidly change the conformation of these multisubunit structures and is necessary for the proper progression of the cell cycle.

Transcriptome profiling of histone writers/erasers enzymes across spermatogenesis, mature sperm and pre-cleavage embryo: Implications in paternal epigenome transitions and inheritance mechanisms

“…To analyze the expression profiles of writers and erasers families for histone K acetylation and methylation across spermatogenesis, we curated RNA-seq data available in purified male germ cells from adult mice and selected the dataset GSE162740 (Mayorek et al, 2022). This dataset contains the mRNA profiles of 6 populations of germ cells isolated in triplicates: undifferentiated (SGund) and differentiated spermatogonia (SGdiff) (spermatogonial phase), primary spermatocytes, including pre-leptotene (PreL), leptotene/zygotene (LZ) and pachytene/diplotene (PD) stages (meiosis), and round spermatids (RStid) (spermiogenesis phase).…”

“…FASTQ files were downloaded from NCBI GEO and ArrayExpress databases. Transcriptomic data of male germ cells was obtained from GSE162740, consisting of germ cells isolated from “Stra8-Tom” mice obtained from the cross of Stra8-cre mice with CAG-LSLtdTomato mice (Mayorek et al, 2022). The gradient expression of the Tomato transgene under Stra8 expression profile, combined with immunostaining for cKit to further discriminate the SGund and SGdiff spermatogonia, was used to FACS sort 6 populations of germ cells in triplicates, and sequence in an Illumina HiSeq 2500 platform.…”

Transcriptome profiling of histone writers/erasers enzymes across spermatogenesis, mature sperm and pre-cleavage embryo: Implications in paternal epigenome transitions and inheritance mechanisms

“…FASTQ files were downloaded from NCBI GEO and ArrayExpress databases. Transcriptomic data of male germ cells was obtained from GSE162740, consisting of germ cells isolated from "Stra8-Tom" mice obtained from the cross of Stra8-cre mice with CAG-LSLtdTomato mice (Mayorek et al, 2022). The gradient expression of the Tomato transgene under Stra8 expression profile, combined with immunostaining for cKit to further discriminate the SGund and SGdiff spermatogonia, was used to FACS sort six populations of germ cells in triplicates, and sequence in an Illumina HiSeq 2,500 platform.…”

“…To analyze the expression profiles of writers and erasers families for histone K acetylation and methylation across spermatogenesis, we curated RNA-seq data available in purified male germ cells from adult mice and selected the dataset GSE162740 (Mayorek et al, 2022). This dataset contains the mRNA profiles of six populations of germ cells isolated in triplicates: undifferentiated Kit-negative (SGund) and differentiated Kit-positive spermatogonia (SGdiff) (spermatogonial phase), primary spermatocytes, including preleptotene (PreL), leptotene/zygotene (LZ) and pachytene/ diplotene (PD) stages (meiosis), and round spermatids (RStid) (spermiogenesis phase).…”