D'Hulst Charlotte, Mina Raena B, Gershon Zachary, Jamet Sophie, Cerullo Antonio, Tomoiaga Delia, Bai Li, Belluscio Leonardo, Rogers Matthew E, Sirotin Yevgeniy, Feinstein Paul
Department of Biological Sciences, Hunter College, City University of New York, New York, NY 10065, USA; The Graduate Center Biochemistry, Biology and Biopsychology and Behavioral Neuroscience Programs, City University of New York, New York, NY 10065, USA.
Developmental Neural Plasticity Section, National Institute of Neurological Disorders and Stroke, NIH, Bethesda, MD 20892, USA.
Cell Rep. 2016 Jul 26;16(4):1115-1125. doi: 10.1016/j.celrep.2016.06.047. Epub 2016 Jul 7.
Typically, ∼0.1% of the total number of olfactory sensory neurons (OSNs) in the main olfactory epithelium express the same odorant receptor (OR) in a singular fashion and their axons coalesce into homotypic glomeruli in the olfactory bulb. Here, we have dramatically increased the total number of OSNs expressing specific cloned OR coding sequences by multimerizing a 21-bp sequence encompassing the predicted homeodomain binding site sequence, TAATGA, known to be essential in OR gene choice. Singular gene choice is maintained in these "MouSensors." In vivo synaptopHluorin imaging of odor-induced responses by known M71 ligands shows functional glomerular activation in an M71 MouSensor. Moreover, a behavioral avoidance task demonstrates that specific odor detection thresholds are significantly decreased in multiple transgenic lines, expressing mouse or human ORs. We have developed a versatile platform to study gene choice and axon identity, to create biosensors with great translational potential, and to finally decode human olfaction.
通常，主嗅觉上皮中约0.1%的嗅觉感觉神经元（OSN）以单一方式表达相同的气味受体（OR），并且它们的轴突在嗅球中聚集成同型肾小球。在这里，我们通过将包含预测的同源结构域结合位点序列TAATGA的21个碱基对序列多聚化，显著增加了表达特定克隆OR编码序列的OSN总数，已知TAATGA在OR基因选择中至关重要。在这些“MouSensors”中维持了单一基因选择。通过已知的M71配体对气味诱导反应进行的体内突触pH荧光蛋白成像显示，M71 MouSensor中功能性肾小球激活。此外，一项行为回避任务表明，在表达小鼠或人类OR的多个转基因品系中，特定气味检测阈值显著降低。我们开发了一个通用平台，用于研究基因选择和轴突身份，创建具有巨大转化潜力的生物传感器，并最终解码人类嗅觉。
