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This study was performed to analyze the developmental changes in bladder response to cholinergic stimulation in detail, highlighting calcium sensitization (CS) and its related pathways. Rats were divided into three groups in accordance with reported time of developmental milestones (newborns, days 1–4; youngsters, days 5–14; and grown-ups, days 15–28). Following cholinergic stimulation (carbachol, 5 µM), the contractile response to detrusor was analyzed with respect to three phases (initial phasic, tonic, and superimposed phasic contractions). Contractile responses were analyzed by their dynamic and kinetic aspects. The responses were further compared in varying external calcium concentrations and in the presence of inhibitors of protein kinase C (PKC) and Rho kinase (ROCK), which are involved in CS. The responses of newborns contrasted with the others by their short and brisk initial phasic contractions, prominent tonic contractions, and delayed participation of irregular superimposed phasic contractions. With development, phasic contractions became prominent, and tonic contractions diminished. These developmental changes in phasic contractions were reproduced when exposed to increasing calcium concentrations. Application of specific inhibitors and molecular phasic analysis revealed that PKC was functional in tonic contractions of the newborns, whereas ROCK took over its role with development. Within a few days of birth, rats’ bladders experienced drastic changes in contractile mechanisms. This included dominance of phasic contractions over tonic contractions due to increased calcium dependence and the maturational shift of the calcium sensitivity mechanism from PKC to ROCK.

Developmental changes in contractile responses to cholinergic stimuli: role of calcium sensitization and related pathways