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Gap junctions in olfactory neurons modulate olfactory sensitivity

机译：嗅觉神经元的缝隙连接调节嗅觉敏感性

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Background One of the fundamental questions in olfaction is whether olfactory receptor neurons (ORNs) behave as independent entities within the olfactory epithelium. On the basis that mature ORNs express multiple connexins, I postulated that gap junctional communication modulates olfactory responses in the periphery and that disruption of gap junctions in ORNs reduces olfactory sensitivity. The data collected from characterizing connexin 43 (Cx43) dominant negative transgenic mice OlfDNCX, and from calcium imaging of wild type mice (WT) support my hypothesis. Results I generated OlfDNCX mice that express a dominant negative Cx43 protein, Cx43/β-gal, in mature ORNs to inactivate gap junctions and hemichannels composed of Cx43 or other structurally related connexins. Characterization of OlfDNCX revealed that Cx43/β-gal was exclusively expressed in areas where mature ORNs resided. Real time quantitative PCR indicated that cellular machineries of OlfDNCX were normal in comparison to WT. Electroolfactogram recordings showed decreased olfactory responses to octaldehyde, heptaldehyde and acetyl acetate in OlfDNCX compared to WT. Octaldehyde-elicited glomerular activity in the olfactory bulb, measured according to odor-elicited c-fos mRNA upregulation in juxtaglomerular cells, was confined to smaller areas of the glomerular layer in OlfDNCX compared to WT. In WT mice, octaldehyde sensitive neurons exhibited reduced response magnitudes after application of gap junction uncoupling reagents and the effects were specific to subsets of neurons. Conclusions My study has demonstrated that altered assembly of Cx43 or structurally related connexins in ORNs modulates olfactory responses and changes olfactory activation maps in the olfactory bulb. Furthermore, pharmacologically uncoupling of gap junctions reduces olfactory activity in subsets of ORNs. These data suggest that gap junctional communication or hemichannel activity plays a critical role in maintaining olfactory sensitivity and odor perception.
机译：背景技术嗅觉中的基本问题之一是嗅觉受体神经元（ORN）是否在嗅觉上皮细胞中表现为独立实体。基于成熟的ORNs表达多种连接蛋白的基础，我推测间隙连接通讯调节周围的嗅觉反应，而ORNs间隙连接的破坏会降低嗅觉敏感性。从表征连接蛋白43（Cx43）显性阴性转基因小鼠OlfDNCX以及从野生型小鼠（WT）钙成像中收集的数据支持了我的假设。结果我产生了OlfDNCX小鼠，该小鼠在成熟的ORN中表达显性负性Cx43蛋白Cx43 /β-gal，以灭活由Cx43或其他与结构相关的连接蛋白组成的间隙连接和半通道。 OlfDNCX的特征表明，Cx43 /β-gal仅在成熟ORN所在的区域表达。实时定量PCR表明，与WT相比，OlfDNCX的细胞机制是正常的。电嗅觉记录显示，与野生型相比，OlfDNCX中对八醛，庚醛和乙酰乙酸酯的嗅觉响应降低。与野生型相比，根据嗅觉引起的肾小球细胞中c-fos mRNA上调测量，嗅觉球中辛醛引起的肾小球活性被限制在OlfDNCX中较小的肾小球层区域。在野生型小鼠中，对八醛敏感的神经元在应用间隙连接解偶联剂后表现出降低的反应幅度，并且其作用特定于神经元的子集。结论我的研究表明，ORNs中Cx43装配的改变或结构相关的连接蛋白可调节嗅觉反应并改变嗅球中的嗅觉激活图。此外，药理学上间隙连接的解偶联降低了ORN子集中的嗅觉活性。这些数据表明，间隙连接通讯或半通道活性在维持嗅觉敏感性和气味感知中起关键作用。

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