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Purpose: A decrease in the human urethral rhabdosphincter is reported with aging due to apoptosis, which may be a cause of urinary incontinence in the elderly population. To explore this mechanism we investigated the effects of tumor necrosis factor-α (Upstate, Temecula, California) on human urethral rhabdosphincter satellite cells. Materials and Methods: Human urethral rhabdosphincter satellite cells were cultured and selected by magnetic affinity cell sorting, extended their life span. Apoptosis induction was examined by flow cytometry and immunocytochemistry. Caspase cascade activation was determined by Western blot analysis. After tumor necrosis factor receptor expression was confirmed we determined the tumor necrosis factor signaling pathway. Results: Tumor necrosis factor-α inhibited human urethral rhabdosphincter satellite cell proliferation. It caused some cells to stain positive for annexin V-fluorescein isothiocyanate but not for propidium iodide, suggesting the induction of early phase apoptosis. Flow cytometry revealed an increased sub-G1 fraction. Western blot analysis showed activation of caspase-8 and 3, and cleavage of poly (adenosine diphosphate-ribose) polymerase. Tumor necrosis factor receptor expression at the mRNA and protein levels was confirmed by reverse transcriptase-polymerase chain reaction and Western blot analysis, respectively. IκBα phosphorylation was noted within 2 to 5 minutes after tumor necrosis factor-α treatment. The tumor necrosis factor-α antagonist etanercept (Wyeth, Collegeville, Pennsylvania) inhibited IκBα activation and reversed tumor necrosis factor-α effects on human urethral rhabdosphincter satellite cells. Conclusions: Since tumor necrosis factor-α induces growth inhibition and apoptosis of human urethral rhabdosphincter satellite cells via tumor necrosis factor receptor activation, it may be involved in age related decreases in the number of human urethral rhabdosphincter cells and be a causative factor for urinary incontinence in the elderly population. [Copyright &y& Elsevier]

Purpose: We clarified the contractile properties of human male periurethral striated muscle fibers to better understand how the rhabdosphincter and the levator ani maintain urinary continence. Materials and Methods: Muscle specimens were obtained from 52 male patients who underwent radical prostatectomy or radical cystectomy. The specimens were frozen in liquid nitrogen. Frozen sections (10 μm thick) were stained with myofibrillar ATPase at different pH values (pH 4.2, 4.6 and 10.6), and evaluated for quantitative parameters and fiber type distribution. Myosin heavy chain analysis was performed using SDS-PAGE. Results: Of all 52 cases 37 provided specimens that could be divided into the 2 major fiber types, type 1 (slow twitch) and type 2 (fast twitch). Although type 1 muscle fibers were predominant in RS and LA muscle groups (RS 69.6 ± 2.7%, LA 67.0 ± 2.0%), mean muscle fiber size was significantly smaller in RS (mean area 906 ± 86 μm2) than in LA (mean area 2,967 ± 170 μm2) (p <0.0001). In 11 specimens type 2 muscle fibers could be subdivided into types 2A (fast fatigue resistant) and 2B (fast fatigable). Type 2A fibers were significantly more prevalent than type 2B fibers (p <0.05). Likewise, MHC analysis of these 11 specimens found a significantly higher percentage of fiber type 2A expression products (MHC 2A) than of fiber type 2B expression products (MHC 2X) (p <0.05). Conclusions: RS and LA contribute to urinary continence mechanism by slow contraction. Moreover, the smaller mean size of muscle fibers in RS suggests more fatigue resistance compared with muscle fibers in LA because small fibers have a shorter diffusion distance for metabolic substrates. These results should help contribute to a more detailed understanding of the function of periurethral striated muscles in the human male. [Copyright &y& Elsevier]