Introduction:

Published data suggest that diabetes mellitus (DM) complications in the lower urinary tract (LUT) may be due to specific malfunctions in signal transduction pathways in LUT. PKC has been recognized as a prominent intracellular signaling pathway that modulates the effects of cholinergic and adrenergic neurotransmission in many tissues/cells. The goal of the current study was to evaluate the changes in profile and translocation of PKC in the bladder of  diabetic rat.

 

Methods:

Detrusor muscle strips from a transgenic rat model of DM (n=15) and similar age-matched control rats (n=3) were isolated in OCT compound after sacrifice. The frozen sections were incubated with the diluted rabbit polyclonal antibody against PKC isoenzymes h, b, and x, Sections were further stained to visualize the isoform of interest (red channel with Cy-3) together with the surface glycoproteins (green channel stained with wheat germ agglutinen conjugated to Oregon Green) and nuclei (blue channel with bisbenzimide). Sections are viewed and photographed with a confocal microscope equipped with fluorescence optics.

 

Results:

1) There is significantly increased presence of PKC bI at extended sarcolemmal contact faces between diabetic detrusor muscle cells and interstitial cells. 2) The presence of PKC bI at the nuclei of detrusor muscle cells do not appear to be different between diabetes and control. 3) In diabetic detrusor muscle cells, PKC h intensity at the cell membrane is decreased compared to the intensity in the cytoplasmic structure. 4) The mean intensity of PKC x in the diabetic diabetic detrusor cell is twice as great as in the control.

 

Conclusions:

1) Isolation and translocation of PKC isoforms in detrusor muscle cells of normal and diabetic rat are reported for the first time; 2) Profile and translocation of PKC isoforms h, b, and x are markedly different in diabetic detrusor muscle cell compared to control.