Investigation of Activated Mouse Olfactory Sensory Neurons via Combined Immunostaining and in situ Hybridization.

Vinicius Miessler de Andrade Carvalho, Thiago Seike Nakahara, Fabio Papes

Journal of visualized experiments : JoVE 2021 Apr 11

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Animals rely on chemical communication to convey and perceive relevant environmental information, ranging from assessment of food quality to detection of available mating partners or threats. In mice, this task is executed primarily by the olfactory system and its underlying subsystems, including the main and accessory olfactory systems. Both have peripheral organs populated by sensory neurons expressing G-protein coupled receptors able to bind chemical cues that reach the nasal cavity. Even though the molecular characteristics of these receptors is well understood, little is known about their cognate specific ligands. The method described here combines in situ hybridization detection of olfactory or vomeronasal receptors with immunodetection of phosphorylated ribosomal protein S6 (pS6) - a marker of neuronal activation. This protocol was devised to identify neurons activated after a single event of exposure to purified or complex chemical stimuli detected by the olfactory organs. Importantly, this technique allows the investigation of neurons triggered in biologically relevant contexts. Ideally, this method should be used to probe the molecular biology of the olfactory system and to study olfactory behaviors.

Citation

Vinicius Miessler de Andrade Carvalho, Thiago Seike Nakahara, Fabio Papes. Investigation of Activated Mouse Olfactory Sensory Neurons via Combined Immunostaining and in situ Hybridization. Journal of visualized experiments : JoVE. 2021 Apr 11(170)