single cell RNA sequencing of adult (P60) vomeronasal organ

The vomeronasal organ of mice consists of two major types of vomeronasal sensory neurons (VSNs) expressing either receptors of the V1R or V2R family. V1R and V2R VSNs form from a common pool of progenitors but have distinct differentiation programs. We analyzed single cell RNA sequencing data of adult VNO and identified differential expression of Notch1 receptor and Dll4 ligand among the neuronal precursors at the VSN dichotomy. We further demonstrated that Notch signaling is required for effective differentiation of V2R+ basal VSNs. Overall design: We employed single-cell RNA sequencing (scRNA-seq) technique to understand the mechanisms underlying cell fate specification of two major types of vomeronasal neurons in mice. As adult neurogenesis happen through out the life at marginal zones of the vomeronasal sensory epithelium in mice, we chose postnatal day 60 to do scRNA-seq in order to collect cells ranging from neuronal progenitors, to immature and mature vomeronasal neurons and non-neuronal cells of both sensory and non-sensory epithelium. The frozen single cell suspension was sent to Singulomics for high-throughput single-cell gene expression profiling using the 10x Genomics Chromium Platform. Contributor: Singulomics Corporation, Bronx, NY

小鼠犁鼻器（vomeronasal organ, VNO）主要包含两类犁鼻感觉神经元（vomeronasal sensory neurons, VSNs），分别表达V1R家族或V2R家族的受体。V1R型与V2R型VSNs起源于同一祖细胞池，但二者拥有截然不同的分化程序。我们对成年小鼠的VNO开展单细胞RNA测序分析，在VSNs分化的二分节点处的神经元前体细胞中，鉴定出Notch1受体与Dll4配体的差异表达特征。我们进一步证实，Notch信号通路对于V2R阳性基底VSNs的有效分化不可或缺。

总体实验设计：本研究采用单细胞RNA测序（single-cell RNA sequencing, scRNA-seq）技术，旨在解析小鼠两类主要犁鼻神经元的细胞命运决定机制。鉴于小鼠犁鼻感觉上皮的边缘区域终身存在成体神经发生过程，我们选择出生后第60天（postnatal day 60, P60）进行scRNA-seq，以收集涵盖神经元祖细胞、未成熟及成熟犁鼻感觉神经元，以及感觉上皮与非感觉上皮的非神经元细胞在内的各类细胞。将制备好的冷冻单细胞悬液送至Singulomics公司，依托10x Genomics Chromium平台完成高通量单细胞基因表达谱分析。

贡献单位：纽约州布朗克斯区Singulomics公司。