Notch3 null mutation in mice causes muscle hyperplasia by repetitive muscle regeneration

Satellite cells are skeletal muscle stem cells responsible for growth, maintenance, and repair of postnatal skeletal muscle. Although several studies have demonstrated that Notch signaling plays a critical role in muscle regeneration through promoting proliferation and self-renewal of satellite cells, the function of Notch3 is yet to be elucidated. We analyzed muscle regeneration in Notch3-deficient mutant mice. We found a remarkable overgrowth of muscle mass in the Notch3-deficient mice but only when they suffered repetitive muscle injuries. Immunochemical analysis found that Notch3 was expressed in Pax7(+)/MyoD(-) quiescent satellite cells and also in Pax7(+)/MyoD(+)-activated satellite cells, but the expression was restricted to around half the population of each cell type. In Notch3-deficient mice, the number of sublaminar quiescent satellite cells was significantly increased compared with those in control mice. We also found that primary cultured myoblasts isolated from the Notch3-deficient mice proliferated faster than those from control mice. Analysis of cultured myofibers revealed that the number of self-renewing Pax7-positive satellite cells attached to the myofiber was increased in the Notch3-deficient mice when compared with control mice. The data obtained in this study suggested that Notch3 pathway might be distinct from Notch1 in muscle regeneration. Because overexpression of Notch3 activated the expression of Nrarp, a negative feedback regulator of Notch signaling, Notch3 might act as a Notch1 repressor by activating Nrarp.