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Modulation of contractile protein troponin-T in chick myocardial cells by basic fibroblast growth factor and
C Vélez1, A E Aránega, C Melguizo
1Department of Morphological Sciences, School of Medicine, University of Granada, Spain.
Insights
Basic fibroblast growth factor (bFGF) significantly increased cardiac troponin-T (TnT) in early-stage chick heart cells. This suggests bFGF promotes cardiomyocyte differentiation during embryonic development.
Area of Science:
- Cardiology
- Developmental Biology
- Molecular Biology
Background:
- Cardiac troponin-T (TnT) is a key contractile protein in cardiomyocytes.
- Growth factors play crucial roles in embryonic development and cell differentiation.
- Understanding molecular markers of cardiomyocyte differentiation is essential.
Purpose of the Study:
- To quantify the effects of basic fibroblast growth factor (bFGF) and platelet-derived growth factor (PDGF) on TnT expression in chick embryo cardiomyocytes.
- To investigate the impact of these growth factors on TnT gene transcription at different embryonic developmental stages.
- To determine the role of bFGF in early cardiomyocyte differentiation.
Main Methods:
- Primary cultures of chick embryo cardiomyocytes from Hamburger and Hamilton (HH) stages 19, 29, and 39.
- Treatment with bFGF and PDGF.
- Analysis of TnT expression using Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting.
- Quantification of TnT mRNA levels.
Main Results:
- Cytoplasmic and cytoskeletal TnT concentrations were dependent on embryonic stage and growth factor treatment.
- bFGF significantly increased cytoplasmic TnT expression (p < 0.001) and TnT mRNA in HH stage 19 cardiomyocytes.
- bFGF induced less pronounced changes in TnT accumulation at HH stage 39 compared to controls.
Conclusions:
- bFGF plays a significant role in promoting cardiomyocyte differentiation during early embryonic development.
- The effects of bFGF on TnT expression are stage-dependent.
- TnT expression serves as a marker for bFGF-mediated cardiomyocyte differentiation.
Abstract:
We quantified the effect of basic fibroblast growth factor (bFGF) and platelet-derived growth factor (PDGF) on the contractile protein troponin-T (TnT) at the cellular and subcellular level in cultures of chick embryo cardiomyocytes obtained from Hamburger and Hamilton's (HH) stage 19, 29, and 39 embryos. Because expression of thin-filament molecules is considered a good marker of differentiation in muscle cell cultures, we analyzed the modifications these growth factors induced in the transcription of the gene for chick cardiac TnT. Sodium dodecyl sulfate-polyacrylamate gel electrophoresis (SDS-PAGE) and immunoblotting showed that cytoplasmic and cytoskeletal concentrations of TnT are dependent on the stage of embryonic development analyzed and on the type of growth factors added to the culture. The most significant finding was the increase in TnT expression in the cytoplasmic compartment (p < 0.001), accompanied by a slight increase in TnT mRNA, after treatment with bFGF of chick heart cells obtained at HH stage 19. At HH stage 39, bFGF induced less marked changes in the accumulation of TnT in comparison with untreated cardiomyocytes. Our findings support the hypothesis that bFGF plays a role in cardiomyocyte differentiation during early stages of development.