The effect of the modulation of L2-hydroxyglutarate content in pachytene-diplotene cells of male mice

We found that testis of male mice contain high concentrations of L2-hydroxyglutarate (L2HG) specifically in germ cells at pachytene/ diplotene (PD) stage of spermatogenesis. In order to elucidate the function of 2HG we decreased its level by incubating PD cells with 24 mM oxamate and to rescue the putative phenotype by incubation with 24 mM oxamate together with 0.3 mM cell permeable L2HG. Overall design: PD cells were isolated by sorting of enzyme-digested testicular cells from transgenic mice expressing tdTomato under germ cells specific promotor. PD cells were incubated for 48 hrs with 24 mM oxamate or with 24 mM oxamate together with 0.3 mM octyl L-2HG.