Abstract
Cells respond to their environment by relaying mechanical force into biochemical stimuli that activate intracellular signal transduction pathways. Subjecting cells to in vitro mechanical stretch can mimic cellular responses to changes in the rigidity of the extracellular matrix. Here we describe an in vitro model system that mimics stretch overload in vivo. In this stretch-mediated hypertrophy model, adult rat cardiomyocytes attached to laminin-coated flexible membranes are subjected to cyclic mechanical stretch at an extension level of 10 % at 30 cycles/min. At various time points VEGF secretion into the media is collected and quantitated.
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References
Leychenko A, Konorev E, Jijiwa M et al (2011) Stretch-induced hypertrophy activates NFkB-mediated VEGF secretion in adult cardiomyocytes. PLoS One 6(12), e29055
Shyu KG, Wang B-W, Lin C-M et al (2010) Cyclic stretch enhances the expression of toll-like receptor 4 gene in cultured cardiomyocytes via p38 MAP kinase and NF-kappaB pathway. J Biomed Sci 17:15
Roca-Cusachs P, Iskratsch T, Sheetz MP (2012) Finding the weakest link: exploring integrin-mediated mechanical molecular pathways. J Cell Sci 125(Pt 13):3025–3038
Bernardo BC, Weeks KL, Pretorius L et al (2010) Molecular distinction between physiological and pathological cardiac hypertrophy: experimental findings and therapeutic strategies. Pharmacol Ther 128(1):191–227
Balakumar P, Jagadeesh G (2010) Multifarious molecular signaling cascades of cardiac hypertrophy: can the muddy waters be cleared? Pharmacol Res 62(5):365–383
Honsho S, Nishikawa S, Amano K et al (2009) Pressure-mediated hypertrophy and mechanical stretch induces IL-1 release and subsequent IGF-1 generation to maintain compensative hypertrophy by affecting Akt and JNK pathways. Circ Res 105(11):1149–1158
Sadoshima J, Izumo S (1993) Mechanical stretch rapidly activates multiple signal transduction pathways in cardiac myocytes: potential involvement of an autocrine/paracrine mechanism. EMBO J 12(4):1681–1692
Sadoshima J, Izumo S (1993) Mechanotransduction in stretch-induced hypertrophy of cardiac myocytes. J Recept Res 13(1-4):777–794
Brancaccio M, Hirsch E, Notte A et al (2006) Integrin signalling: the tug-of-war in heart hypertrophy. Cardiovasc Res 70(3):422–433
Ross RS (2002) The extracellular connections: the role of integrins in myocardial remodeling. J Card Fail 8(6 Suppl):S326–S331
Ross RS, Pham C, Shai SY et al (1998) Beta1 integrins participate in the hypertrophic response of rat ventricular myocytes. Circ Res 82(11):1160–1172
Aikawa R, Nagai T, Kudoh S et al (2002) Integrins play a critical role in mechanical stress-induced p38 MAPK activation. Hypertension 39(2):233–238
Lal H, Verma SK, Smith M et al (2007) Stretch-induced MAP kinase activation in cardiac myocytes: differential regulation through beta1-integrin and focal adhesion kinase. J Mol Cell Cardiol 43(2):137–147
Seko Y, Seko Y, Takahashi N et al (1999) Pulsatile stretch stimulates vascular endothelial growth factor (VEGF) secretion by cultured rat cardiac myocytes. Biochem Biophys Res Commun 254(2):462–475
Zheng W, Seftor EA, Meininger CJ et al (2001) Mechanisms of coronary angiogenesis in response to stretch: role of VEGF and TGF-beta. Am J Physiol Heart Circ Physiol 280(2):H909–H917
Piper HM, Probst I, Schwartz T et al (1982) Culturing of calcium stable adult cardiac myocytes. J Mol Cell Cardiol 14(7):397–412
Acknowledgment
We thank Anna Leychenko for her technical expertise in isolating adult rat cardiomyocytes and her help with the cyclic stretch assays and VEGF analysis. M.L. Matter is supported by a grant from the National Institutes of Health (RO1GM104980).
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Matter, M.L. (2015). Induction of VEGF Secretion in Cardiomyocytes by Mechanical Stretch. In: Fiedler, L. (eds) VEGF Signaling. Methods in Molecular Biology, vol 1332. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2917-7_5
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DOI: https://doi.org/10.1007/978-1-4939-2917-7_5
Publisher Name: Humana Press, New York, NY
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