Bladder Dysfunction in an Obese Zucker Rat: The Role of TRPA1 Channels, Oxidative Stress, and Hydrogen Sulfide

Figure 2

Increased TRPA1 expression in the OZR bladder. TRPA1 expression in representative detrusor muscle transverse sections from a total of 5–6 lean Zucker rat (LZR) and obese Zucker rat (OZR) bladders. Detrusor overall innervation was visualized using the general nerve marker PGP 9.5 (green areas) (a and e). Bladder TRPA1 channel immunofluorescence from the LZR (B) and OZR (f) reveals immunopositive nerve trunks (red areas), running parallel to the smooth muscle bundles. Same fields (a, b, e, and f). Immunofluorescence double labelling for the PGP 9.5 and TRPA1 channel in the smooth muscle, demonstrating neuronal colocalization (yellow areas) (c and g). Cell nuclei were counterstained with DAPI (blue areas) (d and h). Scale bars indicate 25 μm. Negative controls showing the lack of immunoreactivity in sections incubated in the absence of the primary antibody (i and j) (). Comparison of the fluorescence density of nerve fibers and TRPA1 in the LZR and OZR, by using ImageJ, showing a higher expression of TRPA1 in nerve fibers of the OZR bladder (k). Western blot of detrusor smooth muscle membranes from the LZR and OZR incubated with the TRPA1 antibody showing a 131 kDa major band, which corresponded to the expected molecular weight, indicating an increased TRPA1 protein expression in the OZR bladder (l) (). vs. the LZR value, unpaired Student’s -test.