Rapid Dissection and Dissociation of the Mouse Olfactory Epithelium for Single-Nucleus Suspensions
Benjamin D W Belfort1, Anthony M Insalaco2, Claude C Chew3
1Genetics and Genomics Graduate Program, Baylor College of Medicine; Department of Molecular and Human Genetics, Baylor College of Medicine; Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital; Medical Scientist Training Program, Baylor College of Medicine.
Abstract:
The murine olfactory epithelium is the initial entry point of the olfactory system, housing various cell types that include olfactory sensory neurons, their regenerating progenitors, and support cells. Olfactory sensory neurons transduce chemical odorants into neural signals, yet the mechanisms underlying how these cells develop and turnover, create synapses with the olfactory bulb, and regulate their odorant receptors remain areas of intense study. Located on the dorsal aspect of the nasal cavity, the olfactory epithelium adheres to intricate bony structures known as turbinates. This anatomy poses unique challenges for its extraction and dissociation, especially in the context of preparing viable single-cell suspensions. Because single-cell suspension protocols often involve preparatory steps (e.g., papain dissociation, FACS) that can stress cell viability and/or delay library preparation, minimizing tissue extraction time is crucial. This article presents a streamlined method for the rapid dissection of the olfactory epithelium and a protocol for generating high-quality single-nucleus suspensions.


