Bressel Olaf Christian, Khan Mona, Mombaerts Peter
Max Planck Research Unit for Neurogenetics, 60438, Frankfurt, Germany.
J Comp Neurol. 2016 Jan 1;524(1):199-209. doi: 10.1002/cne.23835. Epub 2015 Jul 16.
Chemosensory specificity in the main olfactory system of the mouse relies on the expression of ∼1,100 odorant receptor (OR) genes across millions of olfactory sensory neurons (OSNs) in the main olfactory epithelium (MOE), and on the coalescence of OSN axons into ∼3,600 glomeruli in the olfactory bulb. A traditional approach for visualizing OSNs and their axons consists of tagging an OR gene genetically with an axonal marker that is cotranslated with the OR by virtue of an internal ribosome entry site (IRES). Here we report full cell counts for 15 gene-targeted strains of the OR-IRES-marker design coexpressing a fluorescent protein. These strains represent 11 targeted OR genes, a 1% sample of the OR gene repertoire. We took an empirical, "count every cell" strategy: we counted all fluorescent cell profiles with a nuclear profile within the cytoplasm, on all serial coronal sections under a confocal microscope, a total of 685,673 cells in 56 mice at postnatal day 21. We then applied a strain-specific Abercrombie correction to these OSN counts in order to obtain a closer approximation of the true OSN numbers. We found a 17-fold range in the average (corrected) OSN number across these 11 OR genes. In the same series of coronal sections, we then determined the total volume of the glomeruli (TGV) formed by coalescence of the fluorescent axons. We found a strong linear correlation between OSN number and TGV, suggesting that TGV can be used as a surrogate measurement for estimating OSN numbers in these gene-targeted strains.
小鼠主嗅觉系统中的化学感应特异性依赖于在主嗅觉上皮(MOE)中数百万嗅觉感觉神经元(OSN)上约1100个气味受体(OR)基因的表达,以及OSN轴突在嗅球中聚集成约3600个肾小球。一种可视化OSN及其轴突的传统方法是通过内部核糖体进入位点(IRES)将OR基因与轴突标记物进行基因标记,该轴突标记物与OR共同翻译。在这里,我们报告了共表达荧光蛋白的15种OR-IRES-标记设计的基因靶向菌株的全细胞计数。这些菌株代表11个靶向OR基因,占OR基因库的1%样本。我们采用了一种经验性的“计数每个细胞”策略:在共聚焦显微镜下,我们对所有连续冠状切片上细胞质内带有核轮廓的所有荧光细胞轮廓进行计数,在出生后第21天对56只小鼠中的685673个细胞进行了计数。然后,我们对这些OSN计数应用了菌株特异性的阿伯克龙比校正,以更接近真实的OSN数量。我们发现这11个OR基因的平均(校正后)OSN数量范围为17倍。在同一系列冠状切片中,我们随后确定了荧光轴突聚结形成的肾小球总体积(TGV)。我们发现OSN数量与TGV之间存在很强的线性相关性,这表明TGV可以用作估计这些基因靶向菌株中OSN数量的替代测量方法。