Department of Neurobiology, Harvard Medical School, Boston, MA 02115.
Division of Newborn Medicine, Department of Medicine, Boston Children's Hospital, Boston, MA 02115.
Proc Natl Acad Sci U S A. 2018 Jan 30;115(5):E1051-E1060. doi: 10.1073/pnas.1717871115. Epub 2018 Jan 17.
Coordinated changes in gene expression underlie the early patterning and cell-type specification of the central nervous system. However, much less is known about how such changes contribute to later stages of circuit assembly and refinement. In this study, we employ single-cell RNA sequencing to develop a detailed, whole-transcriptome resource of gene expression across four time points in the developing dorsal lateral geniculate nucleus (LGN), a visual structure in the brain that undergoes a well-characterized program of postnatal circuit development. This approach identifies markers defining the major LGN cell types, including excitatory relay neurons, oligodendrocytes, astrocytes, microglia, and endothelial cells. Most cell types exhibit significant transcriptional changes across development, dynamically expressing genes involved in distinct processes including retinotopic mapping, synaptogenesis, myelination, and synaptic refinement. Our data suggest that genes associated with synapse and circuit development are expressed in a larger proportion of nonneuronal cell types than previously appreciated. Furthermore, we used this single-cell expression atlas to identify the Prkcd-Cre mouse line as a tool for selective manipulation of relay neurons during a late stage of sensory-driven synaptic refinement. This transcriptomic resource provides a cellular map of gene expression across several cell types of the LGN, and offers insight into the molecular mechanisms of circuit development in the postnatal brain.
基因表达的协调变化是中枢神经系统早期模式形成和细胞类型特化的基础。然而,对于这些变化如何有助于后期的回路组装和细化,人们知之甚少。在这项研究中,我们采用单细胞 RNA 测序技术,针对大脑中一个经历了明确的产后回路发育程序的视觉结构——背外侧膝状核(LGN),在四个时间点开发了一个详细的、全转录组的基因表达资源。这种方法确定了定义主要 LGN 细胞类型的标志物,包括兴奋性中继神经元、少突胶质细胞、星形胶质细胞、小胶质细胞和内皮细胞。大多数细胞类型在发育过程中都表现出显著的转录变化,动态表达参与不同过程的基因,包括视网膜映射、突触形成、髓鞘形成和突触细化。我们的数据表明,与突触和回路发育相关的基因在以前被认为是神经细胞的更大比例的非神经细胞类型中表达。此外,我们使用这个单细胞表达图谱来鉴定 Prkcd-Cre 小鼠品系作为在感觉驱动的突触细化后期选择性操纵中继神经元的工具。这个转录组资源提供了 LGN 中几个细胞类型的基因表达的细胞图谱,并为了解产后大脑中的回路发育的分子机制提供了线索。