Abstract
Recently developed rat heart myocyte cell lines have afforded us the opportunity to evaluate the expression of several transcription factors associated with early cardiac development. These factors include, but are not limi ted to, Nkx-2.5/Csx, MEF-2C and MLP (Muscle LIM Protein). These factors have been shown to be temporally expressed in pre-cardiac mesenchyme coincident with the earliest stages of heart development. Using the BWEM and CLEM myocyte cell lines as models of the embryonic, committed cardiomyocyte, we have evaluated the basal expression levels of these three genes over multiple passages. Both cell lines express these genes, with MEF-2C being the most abundant based on Northern blot hybridization analyses. Interestingly, as these cells increased their passage number, there was a corresponding increase in their basal expression levels. To evaluate potential ‘downstream’ effectors of these genes, we examined the basal expression levels of two cardiac-specific genes cTNC and MLC-2v. Transcript levels for both of these contractile filament genes were elevated with passage, suggestive of a inductive process mediated by one or all of these three transcription factors. Promoter analysis of MLC-2v expression in the CLEM line shows that this increase is transcriptionally-mediated and the lines retain the necessary regulatory factors to maintain and control the transcription of this gene. Analysis of the dynamics of the regulatory role(s) that these three transcription factors play in cardiac development can now be evaluated in a homogeneous, cell culture system.
Recipient of Servier Investigator Award
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsPreview
Unable to display preview. Download preview PDF.
Abbreviations
- MEF:
-
muscle enhancer factor
- MLP:
-
muscle LIM protein
- TNC:
-
Troponin C
- MLC:
-
Myosin light chain
References
Weintraub H: The myoD family and myogenesis: Redundancy, networks and thresholds. Cell 75: 1241–1244, 1993
Olson EN: Regulation of muscle transcription by the MyoD family. Circ Res 72: 1–6, 1993
Edmondson DG, Olson EN: Helix-loop-helix proteins as regulators of muscle-specific transcription. J Biol Chem 268: 755–758, 1993
Fishman MC, Stainier DYR: Cardiovascular development. Prospects for a genetic approach. Circ Res 74: 757–763, 1994
Jaffredo T, Chestier A, Bachnou N, Dieterlen-Liévre F: MC29-immor-talized clonal avian heart cell lines can partially differentiate in vitro. Exp Cell Res 192: 481–491, 1991
Engelmann GL, Birschenall-Roberts MC, Ruscetti FW, Samarel AM: Formation of fetal rat cardiac cell clones by retroviral transformation: Retention of select myocyte characteristics. J Mol Cell Cardiol 25: 197–213, 1993
Antin PB, Ordahl CP: Isolation and characterization of an avian myogenic cell line. Dev Biol 143:111–121, 1991
Delcarpio JB, Lanson NA, Field LJ, Claycomb WC: Morphological characterization of cardiomyocytes isolated from a transplantable cardiac tumor derived from a transgenic mouse atria (AT-1 cells). Circ Res 69: 1591–1600, 1991
Wang Y.-C, Neckelmann N, Mayne A, Herskowitz A, Srinivasan A, Sell KW, Ahmed-Ansari A: Establishment of a human fetal cardiac myocyte cell line. In Vitro Cell Dev Biol 27A: 63–74, 1991
Doetschman T, Schull M, Kier A, Coffin JD: Embryonic stem cell model systems for vascular morphogenesis and cardiac disorders. Hypertension 22: 618–629, 1993
Miller-Hance WC, LaCorbiere M, Fuller SJ, Evans SM, Lyons G, Schmidt C, Robbins J, Chien KR: In vitro chamber specification during embryonic stem cell cardiogenesis. Expression of the ventricular myosin light chain-2 gene is independent of heart tube formation. J Biol Chem 268: 25244–25252, 1993
Edmondson DG, Lyons GE, Martin JF, Olson EN: Mef2 gene expression marks the cardiac and skeletal muscle lineages during mouse embryogenesis. Development 120: 1251–1263, 1994
Arber S, Halder G, Caroni P: Muscle LIM protein, a novel essential regulator of myogenesis, promotes myogenic differentiation. Cell 79: 221–231,1994
Lints TJ, Parsons LM, Hartley L, Lyons I, Harvey RP: Nkx-2.5: A novel murine homeobox gene expressed in early heart progenitor cells and their myogenic descendents. Development 119: 419–431, 1993
Martin JF, Schartz JJ, Olson EN: Myocyte enhancer factor (MEF) 2C: A tissue-restricted member of the MEF-2 family of transcription factors. Proc Natl Acad Sci 90: 5282–5286, 1993
Parmacek MS, Leiden JM: Structure and expression of the murine slow/ cardiac troponin C gene. J Biol Chem 264: 13217–13225, 1989
Iwaki K, Sukhatme VP, Shubeita HE, Chien KR: α- and β-adrenergic stimulation induces distinct patterns of immediate early gene expression in neonatal rat myocardial cells. J Biol Chem 265: 13809–13817, 1990
Lee KJ et al.: Myosin light chain-2 luciferase transgenic mice reveal distinct regulatory programs for cardiac and skeletal muscle specific expression of a single contractile protein gene. J Biol Chem 267: 15875–15885, 1992
Lyons I, Parsons LM, Hartley L, Li R, Andrews J, Robb L, Harvey RP: Myogenic and morphogenetic defects in the heart tubes of murine embryos lacking the homeobox gene Nkx2–5. Gene Develop. In press, 1995
Laverriere AC, Macneill C, Muller C, Poelmann RE, Burch JBE, Evans T: GATA-4/5/6, a superfamily of three transcription factors transcribed in developing heart and gut. J Biol Chem 269: 23177–23184, 1994
Ip HS, Wison DB, Heikinheimo M, Tang Z, Ting C-N, Simon MC, Leiden JM, Parmacek MS: The GATA-4 transcription factor transactivates the cardiac muscle-specific troponin C promoter-enhancer in nonmuscle cells. Mol Cell Biol 14: 7517–7526, 1994
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1996 Kluwer Academic Publishers
About this chapter
Cite this chapter
Engelmann, G.L., Worrell, R.A., Duff, R.A., Grutkoski, P.S., Chien, K.R., Harvey, R.P. (1996). Expression of cardiac muscle markers in rat myocyte cell lines. In: Lamers, J.M.J., Verdouw, P.D. (eds) Biochemistry of Signal Transduction in Myocardium. Developments in Molecular and Cellular Biochemistry, vol 17. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-1275-8_10
Download citation
DOI: https://doi.org/10.1007/978-1-4613-1275-8_10
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4612-8544-1
Online ISBN: 978-1-4613-1275-8
eBook Packages: Springer Book Archive