Abstract:
The skin is the largest sensory organ that is densely innervated by highly specialized sensory neurons allowing the detection of a wide range of stimulations including light touch, temperature, itch and pain. Our knowledge of the sets of genes instructing the functional specialization of sensory neurons is just emerging. In a previous study, we have identified a new Gαi inhibitory interacting protein (GINIP) that marks two distinct subsets of skin-innervating sensory neurons conveying noxious and pleasant touch: the MRGPRD-expressing C-fibers specialized in noxious touch and the TH(+)/TAFA4(+)/V-GLUT3(+) C-Low Threshold MechanoReceptors (C-LTMRs), part of neurons processing pleasant touch. In the recent study published by Reynders et al. (2015), we took advantage of GINIP(mCherry) mouse model in combination with Isolectin B4 (IB4) cell surface labeling and fluorescence activated cell sorting (FACS). We successfully purified MRGPRD(+), C-LTMRs and a heterogeneous population of sensory neurons and subjected their RNA contents RNA-deep sequencing (RNA-seq). The subsequent RNA-seq experiment led to the generation of unique sets of data representative of pure transcriptome profiles of each subset. As a result of this pioneering approach, we established the combinatorial expression of the sets of genes that could dictate the functional specializations of MRGPRD(+) neurons and C-LTMRs. Herein we provide details regarding the experimental design, the quality controls and statistical analysis of the data deposited at Gene Expression Omnibus under the accession number GSE64091.
摘要:
皮肤是最大的感觉器官,由高度专业化的感觉神经元密集支配,允许检测各种刺激,包括轻触,温度,痒和疼痛。我们对指导感觉神经元功能专业化的基因集的知识刚刚出现。在之前的研究中,我们已经确定了一种新的Gαi抑制相互作用蛋白(GINIP),它标志着两个不同的子集的皮肤神经感觉神经元传递有害和愉快的触摸:表达MRGPRD的C纤维,专门用于有害触摸和TH()/TAFA4()/V-GLUT3()C-低阈值机械受体(C-LTs),部分神经元处理愉快的触摸。在Reynders等人最近发表的研究中。(2015),我们利用GINIP(mCherry)小鼠模型结合IsolectinB4(IB4)细胞表面标记和荧光激活细胞分选(FACS)。我们成功纯化了MRGPRD(+),C-LTMR和异质群体的感觉神经元并对其RNA内容物进行RNA深度测序(RNA-seq)。随后的RNA-seq实验导致产生代表每个子集的纯转录组概况的独特数据集。由于这种开创性的方法,我们建立了一组基因的组合表达,这些基因可以决定MRGPRD()神经元和C-LTMR的功能专业化。在这里,我们提供有关实验设计的细节,保藏在基因表达Omnibus上的数据的质量控制和统计分析,登录号为GSE64091。