The Effect of Mechanical Force on Gene Expression of Human Bladder Smooth Muscle Cells

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Callan, Christopher A
Smooth Muscle; Urinary Bladder; Fibrosis; Fibrillar Collagens; Medicine and Health Sciences; Molecular Genetics; Urogenital System
thesis / dissertation description
The purpose of this project is to define, at the molecular level, the process by which gene expression of the extracellular matrix is regulated by mechanical forces in the Human Bladder Smooth Muscle cells (BSMCs). The goal is to first localize several functionally distinct transmembrane proteins; Sarcoglycans (α, β, γ, δ and ε), cytoskeletal proteins Vimentin, and Desmin to verify their presence in the cultured BSMCs using fluorescent-labeled antibodies specific for each protein. The sarcoglycans are primarily responsible for transferring intracellular force generated by the interaction of actin and myosin while the extracellular proteins are responsible for linking the cells to the extracellular matrix. These proteins comprise several collagen matrix proteins, e.g., type I, type III and type IV collagens and an integrin-receptor (11, 64). To evaluate the functions of these proteins, we will subject the BSMCs, using a cell strain apparatus, to a level of force which mimics those experienced by the bladder wall during the filling/emptying cycles. This investigation seeks to find supporting evidence to determine if altered mechanical stimulation of bladder smooth muscle cells results in an up-regulation of type III collagen as well as additional genes which mediate the transfer of intracellular force to the extracellular environment.