Abstract
Aim:
To investigate the effects of the wingless-related MMTV integration site 3A (Wnt3a) signaling on the proliferation, migration, and the myogenic and adipogenic differentiation of rat bone marrow mesenchymal stem cells (rMSC).
Methods:
Primary MSC were isolated and cultured from Sprague-Dawley rats and characterized by flow cytometry. Mouse L cells were transfected with Wnt3a cDNA, and conditioned media containing active Wnt3a proteins were prepared. Cell proliferation was evaluated by cell count and 5-bromodeoxyuridine incorporation assay. The migration of rMSC was performed by using a transwell migration and wound healing assay. The myogenic and adipogenic differentiation in rMSC were examined by light microscopy, immunofluorescence, and RT-PCR at different time points after myogenic or adipogenic introduction.
Results:
Wnt3a signaling induced β-catenin nuclear translocation and activated the Wnt pathway in rMSC. In the presence of Wnt3a, rMSC proliferated more rapidly than the control cells, keeping their differentiation potential. Moreover, Wnt3a signaling induced 2.62% and 3.76% of rMSC-expressed desmin and myosin heavy chain after being cultured in myogenic medium. The myogenic differentiation genes, including Pax7, MyoD, Myf5, Myf4, and myogenin, were activated after Wnt3a treatment. On the other hand, Wnt3a inhibited the adipogenic differentiation in rMSC through the downregulated expression of CCAAT/enh ancer-binding protein alpha (C/EBPalph a) and peroxisome proliferator-activated receptor gamma (PPARgamma). Furthermore, Wnt3a promoted the migration capacity of rMSC.
Conclusion:
The results indicate that Wnt3a signaling can induce myogenic differentiation in rMSC. Wnt3a signaling is also involved in the regulation of the proliferation and migration of rMSC. These results could provide a rational foundation for cell-based tissue repair in humans.
Similar content being viewed by others
Article PDF
References
Friedenstein AJ, Gorskaja JF, Kulagina NN . Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Hematol 1976; 4: 267–74.
Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284: 143–7.
Jiang Y, Jahagirdar BN, Reinhardt RL, Schwartz RE, Keene CD, Ortiz-Gonzalez XR, et al. Pluripotency of mesenchymal stem cells derived from adult marrow. Nature 2002; 418: 41–9.
Reyes M, Lund T, Lenvik T, Aguiar D, Koodie L, Verfaillie CM . Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells. Blood 2001; 98: 2615–25.
Price FD, Kuroda K, Rudnicki MA . Stem cell based therapies to treat muscular dystrophy. Biochim Biophys Acta 2007; 1772: 272–83.
Dezawa M, Ishikawa H, Itokazu Y, Yoshihara T, Hoshino M, Takeda S, et al. Bone marrow stromal cells generate muscle cells and repair muscle degeneration. Science 2005; 309: 314–7.
De Bari C, Dell'Accio F, Vandenabeele F, Vermeesch JR, Raymackers JM, Luyten FP . Skeletal muscle repair by adult human mesenchymal stem cells from synovial membrane. J Cell Biol 2003; 160: 909–18.
Parker MH, Seale P, Rudnicki MA . Looking back to the embryo: defining transcriptional networks in adult myogenesis. Nat Rev Genet 2003; 4: 497–507.
Moon RT, Kohn AD, De Ferrari GV, Kaykas A . WNT and beta-catenin signalling: diseases and therapies. Nat Rev Genet 2004; 5: 691–701.
Yu JM, Kim JH, Song GS, Jung JS . Increase in proliferation and differentiation of neural progenitor cells isolated from postnatal and adult mice brain by Wnt-3a and Wnt-5a. Mol Cell Biochem 2006; 288: 17–28.
Gordon MD, Nusse R . Wnt signaling: multiple pathways, multiple receptors, and multiple transcription factors. J Biol Chem 2006; 281: 22 429–33.
Aberle H, Bauer A, Stappert J, Kispert A, Kemler R . Beta-catenin is a target for the ubiquitin-proteasome pathway. EMBO J 1997; 16: 3797–804.
Li L, Yuan H, Xie W, Mao J, Caruso AM, McMahon A, et al. Dishevelled proteins lead to two signaling pathways. Regulation of LEF-1 and c-Jun N-terminal kinase in mammalian cells. J Biol Chem 1999; 274: 129–34.
Munsterberg AE, Kitajewski J, Bumcrot DA, McMahon AP, Lassar AB . Combinatorial signaling by Sonic hedgehog and Wnt family members induces myogenic bHLH gene expression in the somite. Genes Dev 1995; 9: 2911–22.
Tajbakhsh S, Borello U, Vivarelli E, Kelly R, Papkoff J, Duprez D, et al. Differential activation of Myf5 and MyoD by different Wnts in explants of mouse paraxial mesoderm and the later activation of myogenesis in the absence of Myf5. Development 1998; 125: 4155–62.
Borello U, Coletta M, Tajbakhsh S, Leyns L, De Robertis EM, Buckingham M, et al. Transplacental delivery of the Wnt antagonist Frzb1 inhibits development of caudal paraxial mesoderm and skeletal myogenesis in mouse embryos. Development 1999; 126: 4247–55.
Ikeya M, Takada S . Wnt signaling from the dorsal neural tube is required for the formation of the medial dermomyotome. Development 1998; 125: 4969–76.
Polesskaya A, Seale P, Rudnicki MA . Wnt signaling induces the myogenic specification of resident CD45+ adult stem cells during muscle regeneration. Cell 2003; 113: 841–52.
Neth P, Ciccarella M, Egea V, Hoelters J, Jochum M, Ries C . Wnt signaling regulates the invasion capacity of human mesenchymal stem cells. Stem Cells 2006; 24: 1892–903.
Jian H, Shen X, Liu I, Semenov M, He X, Wang XF . Smad3-dependent nuclear translocation of beta-catenin is required for TGF-beta1-induced proliferation of bone marrow-derived adult human mesenchymal stem cells. Genes Dev 2006; 20: 666–74.
De Boer J, Wang HJ, Van Blitterswijk C . Effects of Wnt signaling on proliferation and differentiation of human mesenchymal stem cells. Tissue Eng 2004; 10: 393–401.
Wakitani S, Saito T, Caplan AI . Myogenic cells derived from rat bone marrow mesenchymal stem cells exposed to 5-azacytidine. Muscle Nerve 1995; 18: 1417–26.
Shibamoto S, Higano K, Takada R, Ito F, Takeichi M, Takada S . Cytoskeletal reorganization by soluble Wnt-3a protein signalling. Genes Cells 1998; 3: 659–70.
Hyatt JP, Roy RR, Baldwin KM, Edgerton VR . Nerve activity-independent regulation of skeletal muscle atrophy: role of MyoD and myogenin in satellite cells and myonuclei. Am J Physiol Cell Physiol 2003; 285: C1161–73.
Armand AS, Pariset C, Laziz I, Launay T, Fiore F, Della Gaspera, B, et al. FGF6 regulates muscle differentiation through a calcineurin-dependent pathway in regenerating soleus of adult mice. J Cell Physiol 2005; 204: 297–308.
Reinecke H, Poppa V, Murry CE . Skeletal muscle stem cells do not transdifferentiate into cardiomyocytes after cardiac grafting. J Mol Cell Cardiol 2002; 34: 241–9.
Hamdi HK, Castellon R . ACE inhibition actively promotes cell survival by altering gene expression. Biochem Biophys Res Commun 2003; 310: 1227–35.
Ross SE, Hemati N, Longo KA, Bennett CN, Lucas PC, Erickson RL, et al. Inhibition of adipogenesis by Wnt signaling. Science 2000; 289: 950–3.
de Boer J, Siddappa R, Gaspar C, van Apeldoorn A, Fodde R, van Blitterswijk C . Wnt signaling inhibits osteogenic differentiation of human mesenchymal stem cells. Bone 2004; 34: 818–26.
Pinto D, Clevers H . Wnt control of stem cells and differentiation in the intestinal epithelium. Exp Cell Res 2005; 306: 357–63.
Nguyen H, Rendl M, Fuchs E . Tcf3 governs stem cell features and represses cell fate determination in skin. Cell 2006; 127: 171–83.
Lie DC, Colamarino SA, Song HJ, Desire L, Mira H, Consiglio A, et al. Wnt signalling regulates adult hippocampal neurogenesis. Nature 2005; 437: 1370–5.
Reya T, Duncan AW, Ailles L, Domen J, Scherer DC, Willert K, et al. A role for Wnt signalling in self-renewal of haematopoietic stem cells. Nature 2003; 423: 409.
Wagner J, Schmidt C, Nikowits W Jr, Christ B . Compartmentalization of the somite and myogenesis in chick embryos are influenced by wnt expression. Dev Biol 2000; 228: 86–94.
Petropoulos H, Skerjanc IS . Beta-catenin is essential and sufficient for skeletal myogenesis in P19 cells. J Biol Chem 2002; 277: 15393–9.
Belema Bedada, F, Technau A, Ebelt H, Schulze M, Braun T . Activation of myogenic differentiation pathways in adult bone marrow-derived stem cells. Mol Cell Biol 2005; 25: 9509–19.
Vertino AM, Taylor-Jones JM, Longo KA, Bearden ED, Lane TF, McGehee, RE, et al. Wnt10b deficiency promotes coexpression of myogenic and adipogenic programs in myoblasts. Mol Biol Cell 2005; 16: 2039–48.
Christodoulides C, Laudes M, Cawthorn WP, Schinner S, Soos M, O'Rahilly S, et al. The Wnt antagonist Dickkopf-1 and its receptors are coordinately regulated during early human adipogenesis. J Cell Sci 2006; 119: 2613–20.
Pinto D, Clevers H . Wnt, stem cells and cancer in the intestine. Biol Cell 2005; 97: 185–96.
Qiang YW, Walsh K, Yao L, Kedei N, Blumberg PM, Rubin JS, et al. Wnts induce migration and invasion of myeloma plasma cells. Blood 2005; 106: 1786–93.
Brabletz T, Hlubek F, Spaderna S, Schmalhofer O, Hiendlmeyer E, Jung A, et al. Invasion and metastasis in colorectal cancer: epithelial-mesenchymal transition, mesenchymal-epithelial transition, stem cells and beta-catenin. Cells Tissues Organs 2005; 179: 56–65.
Reya T, Clevers H . Wnt signalling in stem cells and cancer. Nature 2005; 434: 843–50.
Author information
Authors and Affiliations
Corresponding author
Additional information
Project supported by the National Natural Science Foundation of China (No 30370510 and 30170337) and the Key Project of the State Ministry of Public Health (No 2001321).
Rights and permissions
About this article
Cite this article
Shang, Yc., Wang, Sh., Xiong, F. et al. Wnt3a signaling promotes proliferation, myogenic differentiation, and migration of rat bone marrow mesenchymal stem cells. Acta Pharmacol Sin 28, 1761–1774 (2007). https://doi.org/10.1111/j.1745-7254.2007.00671.x
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1111/j.1745-7254.2007.00671.x
Keywords
This article is cited by
-
Identification of osteogenic progenitor cell-targeted peptides that augment bone formation
Nature Communications (2020)
-
Comprehensive analysis of lncRNAs and mRNAs in skeletal muscle of rainbow trout (Oncorhynchus mykiss) exposed to estradiol
Scientific Reports (2017)
-
Human MSCs promotes colorectal cancer epithelial–mesenchymal transition and progression via CCL5/β-catenin/Slug pathway
Cell Death & Disease (2017)
-
Effect of SKL2001 on the neuronal survival mechanism in Parkinson’s disease
Molecular & Cellular Toxicology (2017)
-
Activation of Wnt3a signaling promotes myogenic differentiation of mesenchymal stem cells in mdx mice
Acta Pharmacologica Sinica (2016)