Perforated Patch-clamp Recording of Mouse Olfactory Sensory Neurons in Intact Neuroepithelium: Functional Analysis of Neurons Expressing an Identified Odorant Receptor.
Analyzing the physiological responses of olfactory sensory neurons (OSN) when stimulated with specific ligands is critical to understand the basis of olfactory-driven behaviors and their modulation. These coding properties depend heavily on the initial interaction between odor molecules and the olfactory receptor (OR) expressed in the OSNs. The identity, specificity and ligand spectrum of the expressed OR are critical. The probability to find the ligand of the OR expressed in an OSN chosen randomly within the epithelium is very low. To address this challenge, this protocol uses genetically tagged mice expressing the fluorescent protein GFP under the control of the promoter of defined ORs. OSNs are located in a tight and organized epithelium lining the nasal cavity, with neighboring cells influencing their maturation and function. Here we describe a method to isolate an intact olfactory epithelium and record through patch-clamp recordings the properties of OSNs expressing defined odorant receptors. The protocol allows one to characterize OSN membrane properties while keeping the influence of the neighboring tissue. Analysis of patch-clamp results yields a precise quantification of ligand/OR interactions, transduction pathways and pharmacology, OSNs' coding properties and their modulation at the membrane level.
UMR Centre des Sciences du Goŭt et de l'Alimentation, CNRS, INRA, Université de Bourgogne.
David
D
Jarriault
UMR Centre des Sciences du Goŭt et de l'Alimentation, CNRS, INRA, Université de Bourgogne; xavier.grosmaitre@u-bourgogne.fr.
Xavier
X
Grosmaitre
D000818:Animals; D015870:Gene Expression; D017930:Genes, Reporter; D051379:Mice; D008822:Mice, Transgenic; D009812:Odorants; D009831:Olfactory Mucosa / Q000378:metabolism / Q000502:physiology*; D018034:Olfactory Receptor Neurons / Q000378:metabolism / Q000502:physiology*; D018408:Patch-Clamp Techniques / Q000379:methods*; D018035:Receptors, Odorant / Q000096:biosynthesis*; D012903:Smell / Q000502:physiology
D018035:Receptors, Odorant