\u3cem\u3eIn vitro\u3c/em\u3e Collagen Gel Model for Tissue Damage: Towards a Study of Dystrophin-Glycoprotein Complex and Reactive Oxygen Species

Abstract

Duchenne\u27s Muscular Dystrophy is caused by a deficiency in the dystrophin protein, a component of the Dystrophin-Glycoprotein Complex (DGC), which serves as a structural link between the sarcolemma and the cytoskeleton. Neuronal Nitric Oxide Synthase is a critical enzyme in the sarcolemma responsible for catalyzing the formation of nitric oxide (NO). This enzyme is an important molecular component of the DGC. We studied the C2C12 pre-myogenic cell line by growing them in 3D collagen gels to form a model for muscle development. This muscle model maintains the cells in tension while they differentiate, and can be compared to cells grown in a stress-free environment as a control. We used the method q-RT-PCR to measure the expression of specific muscle markers in two distinct cellular environments. Histological images allowed us to assess cell morphology within each sample. This study provides preliminary data for future plans to test the effects of NO generated by plasma to answer mechanistic questions such as: 1) do increased levels of NO affect muscle-specific gene expression in the presence and absence of dystrophin, 2) will the increased NO level stabilize the DGC within the cell, and 3) are other types of muscle cells (skeletal, cardiac, and smooth) affected by increased NO in cells