Michigan Neuroscience Institute, University of Michigan, Ann Arbor, MI, 48109, USA.
Department of Ophthalmology, Stanford University School of Medicine, Palo Alto, CA, 94304, USA.
Sci Rep. 2020 Jan 15;10(1):351. doi: 10.1038/s41598-019-57194-0.
Improved in situ hybridization methods for mRNA detection in tissues have been developed based on the hybridization chain reaction (HCR). We show that in situ HCR methods can be used for the detection of microRNAs in tissue sections from mouse retinas. In situ HCR can be used for the detection of two microRNAs simultaneously or for the combined detection of microRNA and mRNA. In addition, miRNA in situ HCR can be combined with immunodetection of proteins. We use these methods to characterize cells expressing specific microRNAs in the mouse retina. We find that miR-181a is expressed in amacrine cells during development and in adult retinas, and it is present in both GABAergic and glycinergic amacrine cells. The detection of microRNAs with in situ HCR should facilitate studies of microRNA function and gene regulation in the retina and other tissues.
基于杂交链式反应（HCR），我们开发了用于检测组织中 mRNA 的改良原位杂交方法。我们证明了原位 HCR 方法可用于检测小鼠视网膜组织切片中的 microRNAs。原位 HCR 可用于同时检测两种 microRNAs，也可用于 microRNA 和 mRNA 的联合检测。此外，miRNA 原位 HCR 可与蛋白质的免疫检测相结合。我们使用这些方法来描绘在小鼠视网膜中表达特定 microRNAs 的细胞。我们发现，miR-181a 在发育过程中以及在成年视网膜的无长突细胞中表达，并且存在于 GABA 能和甘氨酸能无长突细胞中。原位 HCR 检测 microRNAs 应有助于研究视网膜和其他组织中 microRNA 的功能和基因调控。
