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Differentiation-dependent PTPIP51 Expression in Human Skeletal Muscle Cell Culture

Justus Barop, Heinrich Sauer, Klaus Steger and Monika Wimmer

Institute of Anatomy and Cell Biology (JB,MW), Department of Physiology (HS), and Department of Urology and Pediatric Urology (KS), Justus-Liebig University, Giessen, Germany

Correspondence to: Justus Barop, Institute of Anatomy and Cell Biology, Justus-Liebig University, 35385 Giessen, Germany. E-mail: Justus.Barop{at}anatomie.med.uni-giessen.de

Protein tyrosine phosphatase-interacting protein 51 (PTPIP51) expression was analyzed in proliferating and differentiating human myogenic cells cultured in vitro. Satellite cell cultures derived from four different individuals were used in this study. To analyze the expression of PTPIP51, myoblasts were cultured under conditions promoting either proliferation or differentiation. In addition, further differentiation of already-differentiated myobtubes was inhibited by resubmitting the cells to conditions promoting proliferation. PTPIP51 protein and mRNA were investigated in samples taken at defined time intervals by immunostaining, immunoblotting, in situ hybridization, and PCR. Image analyses of fluorescence immunostainings were used to quantify PTPIP51 in cultured myoblasts and myotubes. Myoblasts grown in the presence of epidermal and fibroblast growth factors (EGF and FGF), both promoting proliferation, expressed PTPIP51 on a basic level. Differentiation to multinuclear myotubes displayed a linear increase in PTPIP51 expression. The rise in PTPIP51 protein was paralleled by an augmented expression of muscle-specific proteins, namely, sarcoplasmic reticulum Ca²⁺ ATPase and myosin heavy-chain protein, both linked to a progressive state of myotubal differentiation. This differentiation-induced increase in PTPIP51 was partly reversible by resubmission of differentiated myotubes to conditions boosting proliferation. The results clearly point toward a strong association between PTPIP51 expression and differentiation in human muscle cells. (J Histochem Cytochem 57:425–435, 2009)

Key Words: PTPIP51 • PTP1B • human • muscle cell • proliferation • differentiation • SERCA • myosin heavy chain

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