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VEGF induces stress fiber formation in fibroblasts isolated from dystrophic muscle

  • RESEARCH ARTICLE
  • Published:
Journal of Cell Communication and Signaling Aims and scope

Abstract

Treatment with vascular endothelial growth factor (VEGF) to reduce ischemia and enhance both endogenous muscle repair and regenerative cell therapy in Duchenne muscular dystrophy (DMD) has been widely proposed in recent years. However, the interaction between angiogenesis and fibrosis, a hallmark feature of DMD, remains unclear. To date, it has not been determined whether VEGF exerts a pro-fibrotic effect on DMD-derived fibroblasts, which may contribute to further disease progression. Thus, the purpose of this study was to investigate the effect of exogenous VEGF on fibroblast cultures established from a murine model of DMD. Primary fibroblast cultures were established from gastrocnemius and diaphragm muscles of 10 week-old mdx/utrn+/- mice. Quantitative polymerase chain reaction (qPCR) was employed to assess changes in transcript expression of alpha-smooth muscle actin (Acta2), type-1 collagen (Col1a1), connective tissue growth factor (Ctgf/ccn2) and fibronectin (Fn1). Immunofluorescence and Western blot analysis was further employed to visualize changes in protein expression of alpha-smooth muscle actin (α-SMA), CTGF/CCN2 and fibronectin. mRNA levels of Col1a1, Ctgf/ccn2, and FN did not increase following treatment with VEGF in fibroblasts derived from either diaphragm or gastrocnemius muscles. Acta2 expression increased significantly in diaphragm-derived fibroblasts following treatment with VEGF. Morphological assessment revealed increased stress fiber formation in VEGF-treated fibroblasts compared to the untreated control fibroblasts. The findings from this study suggest that further investigation into the effect of VEGF on fibroblast function is required prior to the utilization of the growth factor as a treatment for DMD.

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Abbreviations

DMD:

Duchenne muscular dystrophy

VEGF:

Vascular endothelial growth factor

TGFβ:

Transforming growth factor beta

Col1a1 :

Type I collagen (mRNA)

αSMA:

Alpha-smooth muscle actin

Acta2 :

Alpha-smooth muscle actin (mRNA)

Ctgf/ccn2 :

Connective tissue growth factor (mRNA)

CTGF/CCN2:

Connective tissue growth factor

Fn1 :

Fibronectin (mRNA)

FN:

Fibronectin

qPCR:

Quantitative polymerase chain reaction

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Acknowledgments

We would like to thank Dr. Robert Grange from Virginia Tech, Virginia for generously providing us with the mdx/utrn+/- mice. We would also like to acknowledge Terrie Ann and Dale Forder at the St. Joseph’s Animal Care Facility for assistance with animal care and Dr. Justin Crawford for technical assistance.

Author contributions

KG carried out the experimental work and performed the statistical analysis. KG and LH conceived and designed the study and drafted the manuscript. Both authors read and approved the final manuscript.

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Authors and Affiliations

  1. Lawson Health Research Institute, London, Ontario, Canada

    Kelly M. Gutpell & Lisa M. Hoffman

  2. Department of Anatomy and Cell Biology, Western University, London, Ontario, Canada

    Kelly M. Gutpell & Lisa M. Hoffman

  3. Department of Medical Biophysics, Western University, London, Ontario, Canada

    Kelly M. Gutpell & Lisa M. Hoffman

  4. Imaging Program, Lawson Health Research Institute, 268 Grosvenor Street, London, Ontario, Canada, N6A 4V2

    Lisa M. Hoffman

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  1. Kelly M. Gutpell
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Correspondence to Lisa M. Hoffman.

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Gutpell, K.M., Hoffman, L.M. VEGF induces stress fiber formation in fibroblasts isolated from dystrophic muscle. J. Cell Commun. Signal. 9, 353–360 (2015). https://doi.org/10.1007/s12079-015-0300-z

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  • DOI: https://doi.org/10.1007/s12079-015-0300-z

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