Visualizing sensory transmission between dorsal root ganglion and dorsal horn neurons in co-culture with calcium imaging.
Sensory information is conveyed to the central nervous system by primary afferent neurons within dorsal root ganglia (DRG), which synapse onto neurons of the dorsal horn of the spinal cord. This synaptic connection is central to the processing of both sensory and pain stimuli. Here, we describe a model system to monitor synaptic transmission between DRG neurons and dorsal horn neurons that is compatible with high-throughput screening. This co-culture preparation comprises DRG and dorsal horn neurons and utilizes Ca(2+) imaging with the indicator dye Fura-2 to visualize synaptic transmission. Addition of capsaicin to co-cultures stimulated DRG neurons and led to activation of dorsal horn neurons as well as increased intracellular Ca(2+) concentrations. This effect was dose-dependent and absent when DRG neurons were omitted from the culture. NMDA receptors are a critical component of synapses between DRG and dorsal horn neurons as MK-801, a use-dependent non-competitive antagonist, prevented activation of dorsal horn neurons following capsaicin treatment. This model system allows for rapid and efficient analysis of noxious stimulus-evoked Ca(2+) signal transmission and provides a new approach both for investigating synaptic transmission in the spinal cord and for screening potential analgesic compounds.
Ohshiro
Hiroyuki
H
Discovery Biology Research, Nagoya Laboratories, Pfizer Global Research and Development, Pfizer Inc., 5-2 Taketoyo, Aichi 470-2393, Japan.
Ogawa
Shinji
S
Shinjo
Katsuhiro
K
D000344:Afferent Pathways / Q000502:physiology; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D018920:Coculture Techniques; D003952:Diagnostic Imaging / Q000379:methods*; D005456:Fluorescent Dyes; D016257:Fura-2; D005727:Ganglia, Spinal / Q000378:metabolism*; D020671:Posterior Horn Cells / Q000378:metabolism*; D051381:Rats; D009435:Synaptic Transmission / Q000502:physiology*
D005456:Fluorescent Dyes; D002118:Calcium; D016257:Fura-2