title: An improved method for in vitro morphofunctional analysis of mouse dorsal root ganglia.
abstract: Sensory neurons in dorsal root ganglia (DRGs) are the first-order neurons along the pathway conveying sensory information from the periphery to the central nervous system. The analysis of the morphological and physiological features of these neurons and their alterations in pathology is the necessary prerequisite to understand pain encoding mechanisms. Here, we describe an in vitro procedure for combined morphofunctional analysis of mouse DRGs. Freshly excised DRGs obtained from adult mice were incubated in collagenase to dissolve the ensheathing connective capsule. The degradation of the connective tissue facilitates both access to the neurons by classical recording glass pipettes and the penetration of primary antibodies for immunohistochemical procedures. The entire DRGs were then imaged using a confocal microscope obtaining a fine 3D representation of their cytoarchitecture without requiring tissue sectioning. Thus, our proposed whole-mount preparation represents a flexible in vitro approach for both functional and phenotypic analysis of DRG neurons by at the same time preserving their neuroanatomical relationships.
authors.affiliation: Department of Veterinary Sciences, University of Turin, Largo Braccini 2, 10095 Grugliasco, TO, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Ciglieri
authors.affiliation: Department of Veterinary Sciences, University of Turin, Largo Braccini 2, 10095 Grugliasco, TO, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Ferrini
authors.affiliation: Department of Veterinary Sciences, University of Turin, Largo Braccini 2, 10095 Grugliasco, TO, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Boggio
authors.affiliation: Department of Veterinary Sciences, University of Turin, Largo Braccini 2, 10095 Grugliasco, TO, Italy. Electronic address: chiara.salio@unito.it.
authors.forename: C
authors.initials: C
authors.lastname: Salio
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D017364:Collagenases / Q000737:chemistry*; D005727:Ganglia, Spinal / Q000166:cytology* / Q000502:physiology*; D007089:Image Enhancement / Q000379:methods; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods; D018408:Patch-Clamp Techniques / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D011984:Sensory Receptor Cells / Q000737:chemistry / Q000166:cytology* / Q000502:physiology*; D013329:Structure-Activity Relationship
chemical_list: D017364:Collagenases
keywords: DRG; Immunohistochemistry; Nociceptors; Patch clamp; Whole-mount