Glia-specific nuclei isolation and single cell transcriptomics inform astrocyte pathology in human temporal lobe epilepsy. Glia-specific nuclei isolation and single cell transcriptomics inform astrocyte pathology in human temporal lobe epilepsy

The role of human glia in many neurological disorders is still poorly understood due to the lack of tools that reliably isolate specific glial subpopulations from bulk tissue, directly from their native niche. To better understand the contributions of glial pathology in human epilepsy, we developed and validated a novel sorting strategy that simultaneously isolates nuclei populations of astrocytes (PAX6+), oligodendroglial progenitors (OPCs) (OLIG2+) and neurons (NEUN+) from non-pathological fresh-frozen human postmortem temporal neocortex brain tissue (TL Control) and then employed it, in combination with single cell RNA-seq, to characterize the cell-type specific transcriptome alterations in epilepsy temporal neocortex derived from fresh surgical material in patients with medically refractory temporal lobe epilepsy (TLE). Overall design: Analysis of cell-type specific transcriptome changes in human temporal lobe epilepsy generated from cell-type-specific nuclei populations and single cells

由于缺乏能够直接从原生微环境中、从整体组织内可靠分离特定胶质细胞亚群的工具，人类神经胶质在多种神经系统疾病中的作用仍未得到充分阐明。为更深入解析人类癫痫中胶质细胞病理的贡献机制，我们开发并验证了一种全新的分选策略：该策略可同时从非病变新鲜冷冻尸检人类颞叶新皮层脑组织（TL对照组）中分离出星形胶质细胞（PAX6+）、少突胶质细胞祖细胞（oligodendroglial progenitors, OPCs）（OLIG2+）以及神经元（NEUN+）的细胞核群体；随后我们将该策略与单细胞RNA测序（single cell RNA-seq）技术结合，对源自药物难治性颞叶癫痫（temporal lobe epilepsy, TLE）患者新鲜手术样本的癫痫相关颞叶新皮层的细胞类型特异性转录组变化进行表征。实验整体设计：基于细胞类型特异性细胞核群体与单细胞，对人类颞叶癫痫的细胞类型特异性转录组变化开展分析。