Figure 4

TRPA1 is not functionally up-regulated in retrogradely labeled cutaneous neurons from glabrous plantar skin following skin-only incision injury. a. Representative brightfield (left), WGA-Alexafluor594 retrograde label (middle), and IB4-FITC (right) images of ipsilateral lumbar 3–5 neurons from incised mice with retrograde label and IB4 staining (20x objective). Neurons that brightly fluoresced with Alexafluor594 at least 2-times the standard deviation above autofluorescence were considered to be positively stained for the retrograde tracer and recorded. IB4-positive neurons were defined by a halo of FITC labeling around the entire perimeter of the somata of small-diameter (<27 μm) neurons. b. Percentage of cutaneous neurons responding to CINN. There is no difference in the percentage of labeled WT neurons responding to 100μM CINN from skin-only incised and sham mice. Neurons were pooled from 3 mice per group. c. Amplitude of responses of cutaneous neurons to CINN. Ipsilateral retrograde-labeled neurons from skin-only incised WT mice respond with the same amplitude of intracellular increase to 100μM CINN as neurons from sham mice. Same neurons as those in Figure 4b. d. Percentage of cutaneous neurons responding to CINN defined by IB4 staining. There is no difference in percentage of labeled neurons responding to 100μM CINN defined by IB4 binding. Same neurons as those in Figure 4b. e. Amplitude of responses of cutaneous neurons responding to CINN defined by IB4 staining. There was no difference in amplitude of responses to 100μM CINN in labeled neurons defined by IB4 staining. Same neurons as those in Figure 4b.