Single-cell RNA sequencing of olfactory bulb dopaminergic neurons

Here we performed single cell RNA sequencing of manually sorted olfactory bulb dopaminergic neurons with the goal of studying the transcriptomic basis of neuronal plasticity. Acute olfactory bulb 300 µm slices were obtained from Dat-Cre/Flox-tdTomato (B6.SJL-Slc6a3tm1.1(cre) Bkmn/J, Jax stock 006660 / B6.Cg– Gt(ROSA)26Sortm9(CAG-tdTomato)Hze, Jax stock 007909) juvenile mice. Single cell suspensions were generated using the Neural Tissue Dissociation Kit – Postnatal Neurons (Miltenyi Biotec. Cat no. 130-094-802). Dopaminergic neurons were identified by red fluorescence and manually sorted into individual wells. Analysis of the effects of the transcriptomic changes relating to neuronal plasticity triggered by sensory deprivation is ongoing. 284 single-cell samples, 11 positive controls, 7 no-cell negative controls and 5 negative controls.

本研究对手工分选的嗅球多巴胺能神经元开展了单细胞RNA测序（single cell RNA sequencing），旨在探究神经元可塑性的转录组学基础。本研究从Dat-Cre/Flox-tdTomato（B6.SJL-Slc6a3tm1.1(cre)Bkmn/J，Jax库存号006660 / B6.Cg–Gt(ROSA)26Sortm9(CAG-tdTomato)Hze，Jax库存号007909）幼年小鼠中获取300 μm厚的急性嗅球薄片。采用美天旎生物（Miltenyi Biotec）的神经组织解离试剂盒——产后神经元（Neural Tissue Dissociation Kit – Postnatal Neurons，货号130-094-802）制备单细胞悬液，随后通过红色荧光标记识别多巴胺能神经元，并将其手工分选至单个培养孔中。目前针对感觉剥夺诱导的神经元可塑性相关转录组变化的效应分析正在进行中。本数据集共包含284个单细胞样本、11个阳性对照、7个无细胞阴性对照以及5个阴性对照。