Fibroblast growth factor 2 and protein kinase C alpha are involved in syndecan-4 cytoplasmic domain modulation of turkey myogenic satellite cell proliferation |
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| Authors: | Song Yan McFarland Douglas C Velleman Sandra G |
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| Institution: | a Department of Animal Sciences, Ohio Agricultural Research and Development Center, The Ohio State University, Wooster, OH 44691, USAb Department of Animal and Range Sciences, South Dakota State University, Brookings, SD 57007, USA |
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| Abstract: | Syndecan-4 core protein is composed of extracellular, transmembrane, and cytoplasmic domains. The cytoplasmic domain functions in transmitting signals into the cell through the protein kinase C alpha (PKCα) pathway. The glycosaminoglycan (GAG) and N-linked glycosylated (N-glycosylated) chains attached to the extracellular domain influence cell proliferation. The current study investigated the function of syndecan-4 cytoplasmic domain in combination with GAG and N-glycosylated chains in turkey muscle cell proliferation, differentiation, fibroblast growth factor 2 (FGF2) responsiveness, and PKCα membrane localization. Syndecan-4 or syndecan-4 without the cytoplasmic domain and with or without the GAG and N-glycosylated chains were transfected or co-transfected with a small interfering RNA targeting syndecan-4 cytoplasmic domain into turkey muscle satellite cells. The overexpression of syndecan-4 mutants increased cell proliferation but did not change differentiation. Syndecan-4 mutants had increased cellular responsiveness to FGF2 during proliferation. Syndecan-4 increased PKCα cell membrane localization, whereas the syndecan-4 mutants decreased PKCα cell membrane localization compared to syndecan-4. However, compared to the cells without transfection, syndecan-4 mutants increased cell membrane localization of PKCα. These data indicated that the syndecan‐4 cytoplasmic domain and the GAG and N-glycosylated chains are critical in syndecan-4 regulating satellite cell proliferation, responsiveness to FGF2, and PKCα cell membrane localization. |
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| Keywords: | BSA bovine serum albumin DMEM Dulbecco's modified eagle medium ECM extracellular matrix FGF2 fibroblast growth factor 2 GAG glycosaminoglycan GAPDH glyceraldehyde-3-phosphate dehydrogenase N-glycosylated N-linked glycosylated PBS phosphate buffered saline pCMS pCMS-EGFP PIP2 phosphatidylinositol 4 5-bisphosphate PKCα protein kinase C alpha S4 wild-type syndecan-4 S4C syndecan-4 without cytoplasmic domain S4C-N0 syndecan-4 without cytoplasmic domain and N-linked glycosylated chains S4C-S0 syndecan-4 without cytoplasmic domain and glycosaminoglycan chains S4C-S0N0 syndecan-4 without cytoplasmic domain N-linked glycosylated chains and glycosaminoglycan chains siRNA small interfering RNA SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis SEM standard error of the mean TBS-T Tris-buffered saline |
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