Calcium imaging of primary canine sensory neurons: Small-diameter neurons responsive to pruritogens and algogens.
INTRODUCTION
Rodent primary sensory neurons are commonly used for studying itch and pain neurophysiology, but translation from rodents to larger mammals and humans is not direct and requires further validation to make correlations.


METHODS
This study developed a primary canine sensory neuron culture from dorsal root ganglia (DRG) excised from cadaver dogs. Additionally, the canine DRG cell cultures developed were used for single-cell ratiometric calcium imaging, with the activation of neurons to the following pruritogenic and algogenic substances: histamine, chloroquine, canine protease-activated receptor 2 (PAR2) activating peptide (SLIGKT), compound 48/80, 5-hydroxytryptamine receptor agonist (5-HT), bovine adrenal medulla peptide (BAM8-22), substance P, allyl isothiocyanate (AITC), and capsaicin.


RESULTS
This study demonstrates a simple dissection and rapid processing of DRG collected from canine cadavers used to create viable primary sensory neuron cultures to measure responses to pruritogens and algogens.


CONCLUSION
Ratiometric calcium imaging demonstrated that small-diameter canine sensory neurons can be activated by multiple stimuli, and a single neuron can react to both a pruritogenic stimulation and an algogenic stimulation.
['Ganchingco|Joy Rachel C|JRC|0000-0001-5946-780X', 'Fukuyama|Tomoki|T|0000-0003-3510-3272', 'Yoder|Jeffrey A|JA|0000-0002-6083-1311', 'Bäumer|Wolfgang|W|0000-0002-4346-5911']
D000818:Animals D002102:Cadaver D002132:Calcium Radioisotopes D002211:Capsaicin D002417:Cattle D002478:Cells, Cultured D002738:Chloroquine D004285:Dogs D005727:Ganglia, Spinal D006632:Histamine D006801:Humans D007796:Laminectomy D057054:Molecular Imaging D018377:Neurotransmitter Agents D010146:Pain D010446:Peptide Fragments D011537:Pruritus D019275:Radiopharmaceuticals D011984:Sensory Receptor Cells D012701:Serotonin D017366:Serotonin Receptor Agonists D013373:Substance P
ratiometric calcium imaging dorsal laminectomy dissection dorsal root ganglia cell culture fura-2AM