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TGF-beta1, -beta2 and -beta3 cooperate to facilitate tubulogenesis in the explanted quail heart
Jennifer S Holifield1, Angela M Arlen, Raymond B Runyan
1Department of Anatomy and Cell Biology and Cardiovascular Center, University of Iowa, Iowa City, IA 52242, USA.
Background:
Transforming growth factor-beta (TGF-beta) isoforms have been implicated as both pro- and anti-angiogenic modulators. In this study we addressed the roles of TGF-beta isoforms on coronary tubulogenesis.
Methods:
Embryonic (E6) quail ventricular specimens were explanted onto collagen gels allowing endothelial cells to migrate and form vascular tubes. Growth factors and/or neutralizing growth factor antibodies were added to the cultures. Endothelial cells were identified using a quail endothelial cell marker, QH1. Image analysis was used to quantify aggregate tube length.
Results:
Addition of any isoform (TGF-beta(1), TGF-beta(2) or TGF-beta(3)) virtually prevented tubulogenesis (>95% inhibition), while stimulation of tubulogenesis occurred by adding neutralizing antibodies to TGF-beta(3), but not to TGF-beta(1) or -beta(2). When all three isoforms were added, tubulogenesis was enhanced, indicating the key role of TGF-beta(3). Documentation of the inhibitory effect of TGF-beta isoforms on tubulogenesis is further supported by our experiments in which the marked enhancement of tube formation by bFGF and VEGF was negated when exogenous TGF-beta(1), -beta(2), or -beta(3) were added to the cultures.
Conclusions:
(1) TGF-beta(1), -beta(2) and -beta(3) each inhibits angiogenesis; (2) cooperation between the three TGF-beta isoforms and other angiogenic factors is essential for the regulation of normal tubulogenesis and (3) the stimulatory effect of VEGF or bFGF on tubulogenesis is negated by exogenous TGB-betas.
Insights
Transforming growth factor-beta (TGF-beta) isoforms inhibit coronary tubulogenesis. However, cooperation among TGF-beta isoforms and other factors is essential for normal blood vessel formation.
Area of Science:
- Cardiovascular Biology
- Angiogenesis Research
- Developmental Biology
Background:
- Transforming growth factor-beta (TGF-beta) isoforms are known modulators of angiogenesis, with roles that can be both pro- and anti-angiogenic.
- The specific functions of TGF-beta isoforms in the context of coronary tubulogenesis have not been fully elucidated.
Purpose of the Study:
- To investigate the distinct and cooperative roles of TGF-beta isoforms (TGF-beta(1), TGF-beta(2), and TGF-beta(3)) in regulating coronary tubulogenesis.
- To determine the impact of TGF-beta isoforms on endothelial cell tube formation in the developing heart.
Main Methods:
- Utilized embryonic quail ventricular explants cultured on collagen gels to model coronary tubulogenesis.
- Administered specific TGF-beta isoforms and/or neutralizing antibodies to assess their effects on endothelial cell migration and tube formation.
- Quantified aggregate tube length using image analysis and identified endothelial cells with the QH1 marker.
Main Results:
- All three TGF-beta isoforms (TGF-beta(1), TGF-beta(2), TGF-beta(3)) individually inhibited tubulogenesis by over 95%.
- Neutralizing antibodies against TGF-beta(3), but not TGF-beta(1) or TGF-beta(2), stimulated tubulogenesis, highlighting TGF-beta(3)'s key inhibitory role.
- The pro-angiogenic effects of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) were abolished by the addition of exogenous TGF-beta isoforms.
Conclusions:
- Each TGF-beta isoform (TGF-beta(1), TGF-beta(2), TGF-beta(3)) independently inhibits angiogenesis.
- Cooperation among TGF-beta isoforms and other angiogenic factors is critical for the regulation of normal coronary tubulogenesis.
- Exogenous TGF-betas counteract the stimulatory effects of VEGF and bFGF on tubulogenesis.
