Low Victoria F, Mombaerts Peter
Max Planck Research Unit for Neurogenetics, Max-von-Laue-Strasse 4, D-60438 Frankfurt, Germany.
Neuroscience. 2017 Mar 6;344:167-177. doi: 10.1016/j.neuroscience.2016.12.044. Epub 2017 Jan 3.
In the mouse, odorant receptor proteins (ORs) are G-protein-coupled receptors expressed in mature olfactory sensory neurons (OSNs) of the main olfactory epithelium (MOE). ORs mediate odorant reception at the level of the OSN cilia. Most of the ∼1100 OR genes in the mouse genome are expressed, at the RNA level, in mature OSNs. The literature on antibodies against ORs is limited, and most reports are with antibodies that are not commercially available. Here we have screened 40 commercial antibodies against human and mouse ORs by immunofluorescence staining of coronal cryosections of the MOE of 21-day-old C57BL/6J mice. Various methods of antigen retrieval were tested. Of the 19 antibodies raised against human ORs, three yielded a consistent immunoreactive signal in the mouse MOE; of these three, two appeared to cross react against one or more, unknown, mouse ORs. Of the 21 antibodies raised against mouse ORs, six yielded a consistent immunoreactive signal in the mouse MOE; of these six, two also stained specific glomeruli in the olfactory bulb. Antibody specificity could be validated with gene-targeted mouse strains in the case of three ORs. The number of OSNs immunoreactive for the MOR28/Olfr1507 antibody is greater in C57BL/6J than in 129S6/SvEvTac wild-type mice. Taken together, our results are encouraging: 20-30% of these commercially available antibodies are informative in immunohistochemical analyses of the mouse MOE. The commercial availability of these antibodies should facilitate the study of OR proteins in the MOE and the olfactory bulb, and the replicability of results in the literature.
在小鼠中，气味受体蛋白（ORs）是在主嗅上皮（MOE）的成熟嗅觉感觉神经元（OSNs）中表达的G蛋白偶联受体。ORs在OSN纤毛水平介导气味接收。小鼠基因组中约1100个OR基因中的大多数在RNA水平上在成熟的OSNs中表达。关于抗ORs抗体的文献有限，大多数报告使用的是无商业销售的抗体。在这里，我们通过对21日龄C57BL/6J小鼠MOE冠状冷冻切片进行免疫荧光染色，筛选了40种针对人和小鼠ORs的商业抗体。测试了各种抗原修复方法。在针对人ORs产生的19种抗体中，有3种在小鼠MOE中产生了一致的免疫反应信号；在这3种抗体中，有2种似乎与一种或多种未知的小鼠ORs发生交叉反应。在针对小鼠ORs产生的21种抗体中，有6种在小鼠MOE中产生了一致的免疫反应信号；在这6种抗体中，有2种还对嗅球中的特定肾小球进行了染色。在三种ORs的情况下，可以用基因靶向小鼠品系验证抗体特异性。MOR28/Olfr1507抗体免疫反应阳性的OSNs数量在C57BL/6J小鼠中比在129S6/SvEvTac野生型小鼠中更多。综上所述，我们的结果令人鼓舞：这些商业抗体中有20%-30%在小鼠MOE的免疫组织化学分析中具有参考价值。这些抗体的商业可得性应有助于MOE和嗅球中OR蛋白的研究，以及文献中结果的可重复性。
