Figure 3

cacna1e expression in the nervous system involved in pain transmission. (A and B) X-Gal staining (blue) of the whole SC from a heterozygous mutant. (A) Dorsal view. (B) Cross-section. (C) IB4 binding (brown signal) was assessed in an X-Gal-stained SC section from a heterozygous mutant. (D) Whole lumbar DRG was stained with X-Gal. (E and F) X-Gal-stained lumbar DRG neurons were further stained with molecular markers. (E) Staining with IB4. Some of the X-Gal-stained neurons are also stained with IB4 (arrows). (F) RNA in situ hybridization with an antisense PPT-A riboprobe. Some of the X-Gal-positive neurons show PPT-A signal (arrows). Neurons labeled with only X-Gal were shown by arrowheads in E and F. (G) X-Gal staining of RVM from a heterozygous mutant. Staining was not observed in the RM. (H) X-Gal staining of PAG from a heterozygous mutant. (Scale bars: 1 mm in A, B, and G; 0.5 mm in D and H; 100 μm in C; 50 μm in E and F.) In situ hybridization experiments of wild-type mouse brain sections using DIG-labeled cacna1e riboprobes were in good agreement with those obtained by X-Gal staining of the heterozygous mutant brain, suggesting the X-Gal staining reflects the expression of cacna1e gene (data not shown).