I am a
Home I AM A Search Login

Papers of the Week


Papers: 25 Feb 2023 - 3 Mar 2023

RESEARCH TYPE:
Basic Science


In Vitro Studies, Molecular/Cellular, Neuroimaging


2023 Feb 10


J Vis Exp


36847407


192

In Vivo Calcium Imaging of Neuronal Ensembles in Networks of Primary Sensory Neurons in Intact Dorsal Root Ganglia.

Authors

Shannonhouse J, Gomez R, Son H, Zhang Y, Kim YS

Abstract

Ca imaging can be used as a proxy for cellular activity, including action potentials and various signaling mechanisms involving Ca entry into the cytoplasm or the release of intracellular Ca stores. Pirt-GCaMP3-based Ca imaging of primary sensory neurons of the dorsal root ganglion (DRG) in mice offers the advantage of simultaneous measurement of a large number of cells. Up to 1,800 neurons can be monitored, allowing neuronal networks and somatosensory processes to be studied as an ensemble in their normal physiological context at a populational level in vivo. The large number of neurons monitored allows the detection of activity patterns that would be challenging to detect using other methods. Stimuli can be applied to the mouse hindpaw, allowing the direct effects of stimuli on the DRG neuron ensemble to be studied. The number of neurons producing Ca transients as well as the amplitude of Ca transients indicates sensitivity to specific sensory modalities. The diameter of neurons provides evidence of activated fiber types (non-noxious mechano vs. noxious pain fibers, Aβ, Aδ, and C fibers). Neurons expressing specific receptors can be genetically labeled with td-Tomato and specific Cre recombinases together with Pirt-GCaMP. Therefore, Pirt-GCaMP3 Ca imaging of DRG provides a powerful tool and model for the analysis of specific sensory modalities and neuron subtypes acting as an ensemble at the populational level to study pain, itch, touch, and other somatosensory signals.