Intrinsic optical signals in the dorsal horn of rat spinal cord slices elicited by brief repetitive stimulation

With repetitive electrical stimulation of the dorsal root (20 Hz for 1 s at C-fibre strength), intrinsic optical signals (IOSs), measured as changes in light transmittance, were recorded in the superficial dorsal horn of rat spinal cord slices using a photodiode array imaging device. The mechanism underlying the induction of IOSs was investigated. IOSs elicited by brief repetitive stimulation persisted for 1-2 min and were decreased by reducing external Cl- concentration or by cation-chloride cotransport inhibitors. Furosemide was most effective whilst bumetanide was least effective among the inhibitors tested. A 1-min elevation of external K+ concentration evoked IOSs in the dorsal horn in the absence of stimulation, and K+ -induced IOSs were inhibited by furosemide. These results suggest that the uptake of excess K+ via the furosemide-sensitive, cation-chloride cotransporters underlies the induction of the IOSs. One-minute exposure to hypotonic solutions, which would cause cell swelling, induced IOSs in the superficial dorsal horn. Whilst osmotic-induced IOSs were not affected by furosemide, they were inhibited by HgCl2 in a 2-mercaptoethanol-sensitive manner. The stimulation-induced IOSs were similarly depressed by HgCl2 . In contrast, voltage-sensitive dye signals and field potentials, evoked by single electrical stimuli, were significantly less affected by HgCl2 . These results suggest that there is a specialized water transport pathway in the superficial dorsal horn, and that IOSs elicited by brief repetitive activation of C-fibres are attributable to cell swelling caused by water influx through this pathway, as an osmotic gradient is established by the uptake of K+ via the furosemide-sensitive cotransporters.