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Silvestre JGO. Role of MuRF1 and MuRF2 on functional and structural aspects of myoblasts and fibroblasts skeletal muscle cells. [Ph.D thesis (Science Morphofunctional)]. São Paulo: Instituto de Ciências Biomédicas, Universidade de São Paulo; 2016. The muscle specific RING finger (MuRF) family has been proposed to perform as a linker for myofibril machinery, including titin with microtubules, intermediate filaments and also by acting as an Atrogene during muscle wasting. In the present study our aim was to verify the role of MuRF1 and MuRF2 during the myogenic differentiation of skeletal muscle and its role on skeletal muscle fibroblasts. Initially, we used MuRF1 and MuRF2 double knockout mice (dKO MuRF1/2) and submitted then to cardiotoxin injection (CTX). After that, we analyzed the regenerative process through histological procedure. Next, using in vitro analyzes, we silenced MuRF1 and MuRF2 expression by siRNA and seeking for myogenic differentiation capacity. Our results suggest that dKO MuRF1/2 caused an important impairment on skeletal muscle regeneration, because 28 days after CTX the skeletal muscle showed positive staining to white adipocytes. Moreover, siRNA on myogenic primary cultures showed that MuRF1 and MuRF2 play an important role in determining the fate decision between adipogenic and myogenic differentiation. The silencing using MuRF1 and MuRF2 siRNA impaired myotube formation and increase the expression of adipogenic markers. Another interesting finding was that skeletal muscle fibroblasts can express MuRF1 and MuRF2, and its silencing by siRNA impairs the migration capacity of fibroblasts. These results demonstrate the importance of MuRF1 and MuRF2 during myogenic differentiation of skeletal muscle and an important role at intracellular coordination of stress fiber formation of skeletal muscle fibroblasts.

Papel do MuRF1 e MuRF2 sobre aspectos estruturais e funcionais de mioblastos e fibroblastos musculares esqueléticos.