Target-Captured mRNA from Murine Olfactory Bulb

Methods Mol Biol. 2023:2710:149-170. doi: 10.1007/978-1-0716-3425-7_12.

Abstract

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.

Keywords: Gene expression analysis; Olfactory bulb; Olfactory receptor; RNA sequencing.

MeSH terms

  • Animals
  • Axons
  • Mice
  • Olfactory Bulb
  • Olfactory Receptor Neurons*
  • RNA, Messenger / genetics
  • Receptors, Odorant* / genetics

Substances

  • Receptors, Odorant
  • RNA, Messenger