Skip to main content Accessibility help

Fibroblast Growth Factor-2 regulates proliferation of cardiac myocytes in normal and hypoplastic left ventricles in the developing chick

  • Angela deAlmeida (a1) and David Sedmera (a1) (a2) (a3)


The developing heart increases its mass predominantly by increasing the number of contained cells through proliferation. We hypothesized that addition of fibroblast growth factor-2, a factor previously shown to stimulate division of the embryonic myocytes, to the left ventricular myocardium in an experimental model of left heart hypoplasia created in the chicken would attenuate phenotypic severity by increasing cellular proliferation. We have established an effective mode of delivery of fibroblast growth factor-2 to the chick embryonic left ventricular myocardium by using adenovirus vectors, which was more efficient and better tolerated than direct injection of recombinant fibroblast growth factor-2 protein. Injection of control adenovirus expressing green fluorescent protein did not result in significant alterations in myocytic proliferation or cell death compared with intact, uninjected, controls. Co-injection of adenoviruses expressing green fluorescent protein and fibroblast growth factor-2 was used for verification of positive injection, and induction of proliferation, respectively. Treatment of both normal and hypoplastic left ventricles with fibroblast growth factor-2 expressing adenovirus resulted in to 2 to 3-fold overexpression of fibroblast growth factor-2, as verified by immunostaining. An increase by 45% in myocytic proliferation was observed following injection of normal hearts, and an increase of 39% was observed in hypoplastic hearts. There was a significant increase in anti-myosin immunostaining in the hypoplastic, but not the normal hearts. We have shown, therefore, that expression of exogenous fibroblast growth factor-2 in the late embryonic heart can exert direct effects on cardiac myocytes, inducing both their proliferation and differentiation. These data suggest potential for a novel therapeutic option in selected cases of congenital cardiac disease, such as hypoplastic left heart syndrome.


Corresponding author

Correspondence to: David Sedmera, Institute of Anatomy, 1st Faculty of Medicine, Charles University in Prague, U Nemocnice 3, 128 00 Prague 2, Czech Republic. Phone:+420 224 965 941; Fax: +420 224 965 770; E-mail:


Hide All
1.Bishop, SP. The myocardial cell: normal growth, cardiac hypertrophy and response to injury. Toxicol Pathol 1990; 18: 438453.
2.Saiki, Y, Konig, A, Waddell, J, Rebeyka, IM. Hemodynamic alteration by fetal surgery accelerates myocyte proliferation in fetal guinea pig hearts. Surgery 1997; 122: 412419.
3.Clark, EB, Hu, N, Frommelt, P, Vandekieft, GK, Dummett, JL, Tomanek, RJ. Effect of increased pressure on ventricular growth in stage 21 chick embryos. Am J Physiol 1989; 257: H55H61.
4.Sedmera, D, Hu, N, Weiss, KM, Keller, BB, Denslow, S, Thompson, RP. Cellular changes in experimental left heart hypoplasia. Anat Rec 2002; 267: 137145.
5.Hefti, MA, Harder, BA, Eppenberger, HM, Schaub, MC. Signaling pathways in cardiac myocyte hypertrophy. J Mol Cell Cardiol 1997; 29: 28732892.
6.Pasumarthi, KB, Field, LJ. Cardiomyocyte cell cycle regulation. Circ Res 2002; 90: 10441054.
7.Sylven, C. Angiogenic gene therapy. Drugs Today (Barc) 2002; 38: 819827.
8.Stern, CD. The chick; a great model system becomes even greater. Dev Cell 2005; 8: 917.
9.Antin, PB, Fallon, JF, Schoenwolf, GC. The chick embryo rules (still)! Dev Dyn 2004; 229: 413.
10.Sedmera, D, Pexieder, T, Rychterova, V, Hu, N, Clark, EB. Remodeling of chick embryonic ventricular myoarchitecture under experimentally changed loading conditions. Anat Rec 1999; 254: 238252.
11.Tobita, K, Keller, BB. Right and left ventricular wall deformation patterns in normal and left heart hypoplasia chick embryos. Am J Physiol Heart Circ Physiol 2000; 279: H959969.
12.Sedmera, D, Cook, AC, Shirali, G, McQuinn, TC. Current issues and perspectives in hypoplasia of the left heart. Cardiol Young 2005; 15: 5672.
13.Itoh, N, Ornitz, DM. Evolution of the Fgf and Fgfr gene families. Trends Genet 2004; 20: 563569.
14.Ornitz, DM. FGFs, heparan sulfate and FGFRs: complex interactions essential for development. Bioessays 2000; 22: 108112.
15.Eswarakumar, VP, Lax, I, Schlessinger, J. Cellular signaling by fibroblast growth factor receptors. Cytokine Growth Factor Rev 2005; 16: 139149.
16.Lynch, P, Lee, TC, Fallavollita, JA, Canty, JM Jr, Suzuki, G. Intracoronary administration of AdvFGF-5 (fibroblast growth factor-5) ameliorates left ventricular dysfunction and prevents myocyte loss in swine with developing collaterals and ischemic cardiomyopathy. Circulation 2007; 116: I71I76.
17.Suzuki, G, Lee, TC, Fallavollita, JA, Canty, JM Jr. Adenoviral gene transfer of FGF-5 to hibernating myocardium improves function and stimulates myocytes to hypertrophy and reenter the cell cycle. Circ Res 2005; 96: 767775.
18.Demiroglu, A, Steer, EJ, Heath, C, et al. The t(8;22) in chronic myeloid leukemia fuses BCR to FGFR1: transforming activity and specific inhibition of FGFR1 fusion proteins. Blood 2001; 98: 37783783.
19.Lee, PL, Johnson, DE, Cousens, LS, Fried, VA, Williams, LT. Purification and complementary DNA cloning of a receptor for basic fibroblast growth factor. Science 1989; 245: 5760.
20.Powers, CJ, McLeskey, SW, Wellstein, A. Fibroblast growth factors, their receptors and signaling. Endocr Relat Cancer 2000; 7: 165197.
21.Detillieux, KA, Sheikh, F, Kardami, E, Cattini, PA. Biological activities of fibroblast growth factor-2 in the adult myocardium. Cardiovasc Res 2003; 57: 819.
22.Speir, E, Tanner, V, Gonzalez, AM, Farris, J, Baird, A, Casscells, W. Acidic and basic fibroblast growth factors in adult rat heart myocytes. Localization, regulation in culture, and effects on DNA synthesis. Circ Res 1992; 71: 251259.
23.Lavine, KJ, Yu, K, White, AC, et al. Endocardial and epicardial derived FGF signals regulate myocardial proliferation and differentiation in vivo. Dev Cell 2005; 8: 8595.
24.Colvin, JS, Feldman, B, Nadeau, JH, Goldfarb, M, Ornitz, DM. Genomic organization and embryonic expression of the mouse fibroblast growth factor 9 gene. Dev Dyn 1999; 216: 7288.
25.Pennisi, DJ, Ballard, VL, Mikawa, T. Epicardium is required for the full rate of myocyte proliferation and levels of expression of myocyte mitogenic factors FGF2 and its receptor, FGFR-1, but not for transmural myocardial patterning in the embryonic chick heart. Dev Dyn 2003; 228: 161172.
26.Parlow, MH, Bolender, DL, Kokan-Moore, NP, Lough, J. Localization of bFGF-like proteins as punctate inclusions in the preseptation myocardium of the chicken embryo. Dev Biol 1991; 146: 139147.
27.Sugi, Y, Sasse, J, Lough, J. Inhibition of precardiac mesoderm cell proliferation by antisense oligodeoxynucleotide complementary to fibroblast growth factor-2 (FGF- 2). Dev Biol 1993; 157: 2837.
28.Jimenez, SK, Sheikh, F, Jin, Y, et al. Transcriptional regulation of FGF-2 gene expression in cardiac myocytes. Cardiovasc Res 2004; 62: 548557.
29.Sheikh, F, Hirst, CJ, Jin, Y, et al. Inhibition of TGFbeta signaling potentiates the FGF-2-induced stimulation of cardiomyocyte DNA synthesis. Cardiovasc Res 2004; 64: 516525.
30.Velez, C, Aranega, AE, Melguizo, C, Fernandez, JE, Prados, J, Aranega, A. Modulation of contractile protein troponin-T in chick myocardial cells by basic fibroblast growth factor and platelet-derived growth factor during development. J Cardiovasc Pharmacol 1994; 24: 906913.
31.Franciosi, JP, Bolender, DL, Lough, J, Kolesari, GL. FGF-2-induced imbalance in early embryonic heart cell proliferation: a potential cause of late cardiovascular anomalies. Teratology 2000; 62: 189194.
32.Mima, T, Ueno, H, Fischman, DA, Williams, LT, Mikawa, T. Fibroblast growth factor receptor is required for in vivo cardiac myocyte proliferation at early embryonic stages of heart development. Proc Natl Acad Sci U S A 1995; 92: 467471.
33.Mikawa, T. Retroviral targeting of FGF and FGFR in cardiomyocytes and coronary vascular cells during heart development. Ann N Y Acad Sci 1995; 752: 506516.
34.Kardami, E, Liu, L, Kishore, S, Pasumarthi, B, Doble, BW, Cattini, PA. Regulation of basic fibroblast growth factor (bFGF) and FGF receptors in the heart. Ann N Y Acad Sci 1995; 752: 353369.
35.Sheikh, F, Fandrich, RR, Kardami, E, Cattini, PA. Overexpression of long or short FGFR-1 results in FGF-2-mediated proliferation in neonatal cardiac myocyte cultures. Cardiovasc Res 1999; 42: 696705.
36.Hamburger, V, Hamilton, HL. A series of normal stages in the development of the chick embryo. J Morphol 1951; 88: 4992.
37.Nesbit, M, Nesbit, HK, Bennett, J, et al. Basic fibroblast growth factor induces a transformed phenotype in normal human melanocytes. Oncogene 1999; 18: 64696476.
38.Kajstura, J, Rota, M, Whang, B, et al. Bone marrow cells differentiate in cardiac cell lineages after infarction independently of cell fusion. Circ Res 2005; 96: 127137.
39.Zhou, M, Sutliff, RL, Paul, RJ, et al. Fibroblast growth factor 2 control of vascular tone. Nat Med 1998; 4: 201207.
40.Kang, J, Gu, Y, Li, P, Johnson, BL, Sucov, HM, Thomas, PS. PDGF-A as an epicardial mitogen during heart development. Dev Dyn 2008; 237: 692701.
41.Lavine, KJ, Schmid, GJ, Smith, CS, Ornitz, DM. Novel tool to suppress cell proliferation in vivo demonstrates that myocardial and coronary vascular growth represent distinct developmental programs. Dev Dyn 2008; 237: 713724.
42.Liechty, KW, Nesbit, M, Herlyn, M, Radu, A, Adzick, NS, Crombleholme, TM. Adenoviral-mediated overexpression of platelet-derived growth factor-B corrects ischemic impaired wound healing. J Invest Dermatol 1999; 113: 375383.
43.Donahue, JK, Heldman, AW, Fraser, H, et al. Focal modification of electrical conduction in the heart by viral gene transfer. Nat Med 2000; 6: 13951398.
44.Christensen, G, Minamisawa, S, Gruber, PJ, Wang, Y, Chien, KR. High-efficiency, long-term cardiac expression of foreign genes in living mouse embryos and neonates. Circulation 2000; 101: 178184.
45.Cheng, G, Litchenberg, WH, Cole, GJ, Mikawa, T, Thompson, RP, Gourdie, RG. Development of the cardiac conduction system involves recruitment within a multipotent cardiomyogenic lineage. Development 1999; 126: 50415049.
46.Orlic, D, Kajstura, J, Chimenti, S, et al. Bone marrow cells regenerate infarcted myocardium. Nature 2001; 410: 701705.
47.Zhang, N, Mustin, D, Reardon, W, et al. Blood-borne stem cells differentiate into vascular and cardiac lineages during normal development. Stem Cells Dev 2006; 15: 1728.
48.Linke, A, Muller, P, Nurzynska, D, et al. Stem cells in the dog heart are self-renewing, clonogenic, and multipotent and regenerate infarcted myocardium, improving cardiac function. Proc Natl Acad Sci U S A 2005; 102: 89668971.
49.Tobita, K, Garrison, JB, Li, JJ, Tinney, JP, Keller, BB. Three-dimensional myofiber architecture of the embryonic left ventricle during normal development and altered mechanical loads. Anat Rec A Discov Mol Cell Evol Biol 2005; 283: 193201.
50.Dow, JK, deVere White, RW. Fibroblast growth factor 2: its structure and property, paracrine function, tumor angiogenesis, and prostate-related mitogenic and oncogenic functions. Urology 2000; 55: 800806.
51.Kumar-Singh, S, Weyler, J, Martin, MJ, Vermeulen, PB, Van Marck, E. Angiogenic cytokines in mesothelioma: a study of VEGF, FGF-1 and -2, and TGF beta expression. J Pathol 1999; 189: 7278.
52.deAlmeida, A, McQuinn, T, Sedmera, D. Increased ventricular preload is compensated by myocyte proliferation in normal and hypoplastic fetal chick left ventricle. Circ Res 2007; 100: 13631370.



Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed