Jiang Yue, Gong Naihua Natalie, Hu Xiaoyang Serene, Ni Mengjue Jessica, Pasi Radhika, Matsunami Hiroaki
Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, USA.
University Program in Genetics and Genomics, Duke University Medical Center, Durham, North Carolina, USA.
Nat Neurosci. 2015 Oct;18(10):1446-54. doi: 10.1038/nn.4104. Epub 2015 Aug 31.
The mammalian olfactory system uses a large family of odorant receptors (ORs) to detect and discriminate amongst a myriad of volatile odor molecules. Understanding odor coding requires comprehensive mapping between ORs and corresponding odors. We developed a means of high-throughput in vivo identification of OR repertoires responding to odorants using phosphorylated ribosome immunoprecipitation of mRNA from olfactory epithelium of odor-stimulated mice followed by RNA-Seq. This approach screened the endogenously expressed ORs against an odor in one set of experiments using awake and freely behaving mice. In combination with validations in a heterologous system, we identified sets of ORs for two odorants, acetophenone and 2,5-dihydro-2,4,5-trimethylthiazoline (TMT), encompassing 69 OR-odorant pairs. We also identified shared amino acid residues specific to the acetophenone or TMT receptors and developed models to predict receptor activation by acetophenone. Our results provide a method for understanding the combinatorial coding of odors in vivo.
哺乳动物的嗅觉系统利用一大类气味受体（ORs）来检测和区分无数种挥发性气味分子。理解气味编码需要对ORs和相应气味进行全面映射。我们开发了一种高通量体内鉴定OR库的方法，该方法利用来自气味刺激小鼠嗅觉上皮的mRNA进行磷酸化核糖体免疫沉淀，随后进行RNA测序，以鉴定对气味有反应的OR库。这种方法在一组实验中，使用清醒且自由活动的小鼠，针对一种气味筛选内源性表达的ORs。结合在异源系统中的验证，我们鉴定出了两种气味剂（苯乙酮和2,5-二氢-2,4,5-三甲基噻唑啉（TMT））的OR集，其中包括69个OR-气味剂对。我们还鉴定出了苯乙酮或TMT受体特有的共享氨基酸残基，并开发了预测苯乙酮激活受体的模型。我们的结果提供了一种理解体内气味组合编码的方法。
