Department of Molecular Genetics and Microbiology, Duke University School of Medicine, Durham, NC, USA.
Department of Neurobiology, Duke Institute for Brain Sciences, Durham, NC, USA.
Methods Mol Biol. 2023;2710:149-170. doi: 10.1007/978-1-0716-3425-7_12.
The external world is perceived via sensory receptors arranged in highly organized systems according to functional strategies, which in turn reflect features of critical importance to both the sense and the animal. Thus describing the receptor organization and functional strategies of olfaction, the sense of smell, is crucial for a concrete understanding of the fundamental principles of the system's architecture. Sensory processing in olfactory systems is organized across olfactory bulb (OB) glomeruli, wherein axons of peripheral sensory neurons expressing the same olfactory receptor (OR) co-terminate to transmit receptor-specific activity to mitral/tufted cells, projection neurons in the olfactory bulb. Understanding how receptors map to glomeruli is therefore critical to understanding olfaction.Here we describe a method to enrich low-abundant OR mRNAs from the mouse OB for spatial transcriptomics to generate high-throughput mapping of receptors to glomeruli [2]. Our method combines sequential sectioning along the anteroposterior, dorsoventral, and mediolateral axes with target capture enrichment sequencing to overcome low-abundance target expression. This strategy spatially mapped 86% of olfactory receptors across the olfactory bulb and uncovered a relationship between olfactory receptor sequence and glomerular position.
外部世界是通过根据功能策略组织起来的高度有序的感觉受体来感知的，而这些感觉受体又反映了对感觉和动物都至关重要的特征。因此，描述嗅觉（嗅觉）的受体组织和功能策略对于具体理解系统结构的基本原理至关重要。嗅觉系统中的感觉处理是跨嗅球（OB）小球组织的，其中表达相同嗅觉受体（OR）的外周感觉神经元的轴突共同终止，将受体特异性活性传递到嗅球中的投射神经元——僧帽细胞和丛状细胞。因此，了解受体如何映射到小球对于理解嗅觉至关重要。在这里，我们描述了一种从小鼠 OB 中富集低丰度 OR mRNA 的方法，用于空间转录组学，以生成受体到小球的高通量映射[2]。我们的方法结合了沿前后、背腹和内外轴的连续切片与靶向捕获富集测序，以克服低丰度靶标表达。该策略在嗅球中空间映射了 86%的嗅觉受体，并揭示了嗅觉受体序列与小球位置之间的关系。
