Figure 2

ChAT knock-down using an antisense oligodeoxynucleotide (AS-ODN) against ChAT, and decreased expression of ChAT in the DRG and spinal cord. (A, B) Down-regulation of ChAT activities by the AS-ODN in a DRG cross-section. Representative immunohistochemistry (A) and scatter diagram of ChAT activities (B) showing down-regulation of ChAT protein by the AS-ODN, but not by the MS-ODN. Immunoreactivity (B) was measured as an arbitrary value by automatic fluorescence microscopy using BZ Image Measurement software (Bio-Zero, Keyence, Tokyo, Japan). The evidence for down-regulation was reproduced in another experiment. (C) Representative pictures of ChAT-immunohistochemistry in the spinal cord treated with AS-ODN or MS-ODN. (D) Schematic diagram showing the region of interest. (E) Down-regulation of ChAT immunoreactivity by AS-ODN in laminae I–III of the dorsal horn. Each data point (average from 8 sections) was calculated by the formula [(signal/area in lamina I–III) - (signal/area in gracile fasciculus regions of white matter)]. Results represent the means ± S.E.M. from 3 separate mice. *p < 0.05 vs. vehicle. (F) Selective down-regulation of ChAT immunoreactivity by the AS-ODN in laminae I–III of the dorsal horn. Each point of data (average from 8 sections) was calculated by the formula [(signal/area in lamina I–III) - (signal/area in gracile fasciculus regions of white matter)]/[(signal/area in motor neurons) - (signal/area in white matter)]. Veh: vehicle, AS: AS-ODN and MS: MS-ODN. Results represent the means ± S.E.M. from 3 separate mice. *:p < 0.05 vs. vehicle. Scale bar = 50 μm for (A).