Localization of Shh and Ptch1 RNA by In Situ Hybridization Remained Unchanged Following CN Injury
The RNA localization of Shh and Ptchl was examined by in situ hybridization in control and CN-injured penes 21 days after CN injury. Shh and Ptchl RNA expression was abundant in the smooth muscle of the corpora cavernosal sinusoidal tissue of both control and CN-injured penes (Fig. 4). A change in RNA localization for Shh and Ptchl was not observed following CN injury.
The Active Form of SHH Protein was Significantly Decreased by Western Analysis Following CN Injury
The precursor (46 kDa) and active (19 kDa) forms of SHH protein were quantified by Western analysis in control and CN-injured penes 21 days after CN injury. The active form of SHH protein was significantly decreased after CN injury (control — 0.572 6 0.02, CN-injured — 0.492 6 0.01, P-value — 0.001, Fig. 5). The precursor form was increased, but there was not enough statistical power for the difference to be significant (control — 0.815 6 0.03, CN-injured — 0.879 6 0.03, P-value — 0.08, Fig. 5).
FIG. 4. In situ hybridization showing the localization of Shh and Ptch1 in control and CN-injured Sprague Dawley rat penes 21 days after CN injury. The distribution of Shh and Ptch1 RNA remained unchanged following CN injury in smooth muscle cells of the corpus cavernosal sinusoidal tissue. Original magnification X400. Arrows indicate Shh and Ptch1 RNA localization.
FIG. 5. Graph (a) and gel (b) showing Western analysis of SHH protein in control (C) and CN-injured (CN) penes 21 days after CN injury. The active form of SHH protein (19 kDa) was significantly decreased after CN injury (P-value = 0.001). The precursor form (46 kDa) was increased but was not statistically significant (P-value = 0.08). SEM for the 19-kDa bands was 0.02, which was too small to print error bars on the graph.