Nervous system reduction in branched-chain amino acid metabolism disrupts hippocampal neurogenesis and memory

To study the role of branched-chain amino acid metabolism in neural stem cells we generated two primary neural stem cell culture lines in which on was deleted for the Ppm1k gene using floxed cells and adenovirus delviered cre-recombinase. Overall design: Neural stem cell cultures were generated from control (wild-type/wild-type) and ppm1k flox/flox mice in triplicate and pooled to make control and ppm1l flox/flox lines. Ppm1k was knocked out of the flox/flox line using adenovirus delivered cre-recombinase. After 68 hours primary cell cultures were isolated and submited for 10X genomic sequencing.

本研究为探究支链氨基酸代谢在神经干细胞中的功能角色，借助floxed细胞（floxed cell）与腺病毒递送的Cre重组酶（Cre-recombinase），构建了两株原代神经干细胞培养系，其中一株的Ppm1k基因被敲除。
实验整体设计如下：分别从野生型（wild-type/wild-type）及Ppm1k flox/flox小鼠中分离原代神经干细胞并进行培养，设置三次生物学重复后混合，分别获得野生型对照株与Ppm1l flox/flox株；随后通过腺病毒递送的Cre重组酶对Ppm1l flox/flox株实施Ppm1k基因敲除。培养68小时后，收集原代细胞培养物并提交至10X基因组测序（10X Genomic Sequencing）平台完成测序。