title: Serotonin potentiates transforming growth factor-beta3 induced biomechanical remodeling in avian embryonic atrioventricular valves.
abstract: Embryonic heart valve primordia (cushions) maintain unidirectional blood flow during development despite an increasingly demanding mechanical environment. Recent studies demonstrate that atrioventricular (AV) cushions stiffen over gestation, but the molecular mechanisms of this process are unknown. Transforming growth factor-beta (TGFβ) and serotonin (5-HT) signaling modulate tissue biomechanics of postnatal valves, but less is known of their role in the biomechanical remodeling of embryonic valves. In this study, we demonstrate that exogenous TGFβ3 increases AV cushion biomechanical stiffness and residual stress, but paradoxically reduces matrix compaction. We then show that TGFβ3 induces contractile gene expression (RhoA, aSMA) and extracellular matrix expression (col1α2) in cushion mesenchyme, while simultaneously stimulating a two-fold increase in proliferation. Local compaction increased due to an elevated contractile phenotype, but global compaction appeared reduced due to proliferation and ECM synthesis. Blockade of TGFβ type I receptors via SB431542 inhibited the TGFβ3 effects. We next showed that exogenous 5-HT does not influence cushion stiffness by itself, but synergistically increases cushion stiffness with TGFβ3 co-treatment. 5-HT increased TGFβ3 gene expression and also potentiated TGFβ3 induced gene expression in a dose-dependent manner. Blockade of the 5HT2b receptor, but not 5-HT2a receptor or serotonin transporter (SERT), resulted in complete cessation of TGFβ3 induced mechanical strengthening. Finally, systemic 5-HT administration in ovo induced cushion remodeling related defects, including thinned/atretic AV valves, ventricular septal defects, and outflow rotation defects. Elevated 5-HT in ovo resulted in elevated remodeling gene expression and increased TGFβ signaling activity, supporting our ex-vivo findings. Collectively, these results highlight TGFβ/5-HT signaling as a potent mechanism for control of biomechanical remodeling of AV cushions during development.
authors.affiliation: Department of Mechanical and Aerospace Engineering, Cornell University, Ithaca, New York, United States of America.
authors.forename: Philip R
authors.initials: PR
authors.lastname: Buskohl
authors.forename: Michelle J
authors.initials: MJ
authors.lastname: Sun
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Sun
authors.forename: Robert P
authors.initials: RP
authors.lastname: Thompson
authors.forename: Jonathan T
authors.initials: JT
authors.lastname: Butcher
mesh_terms: D000818:Animals; D001696:Biomechanical Phenomena / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002642:Chick Embryo; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006343:Heart Septal Defects / Q000196:embryology / Q000473:pathology / Q000503:physiopathology; D006351:Heart Valves / Q000187:drug effects / Q000196:embryology* / Q000473:pathology / Q000502:physiology*; D006801:Humans; D008648:Mesoderm / Q000187:drug effects; D009200:Myocardial Contraction / Q000187:drug effects; D010641:Phenotype; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D044403:Receptor, Serotonin, 5-HT2B / Q000378:metabolism; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000378:metabolism; D012701:Serotonin / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D051785:Smad Proteins / Q000378:metabolism; D053782:Transforming Growth Factor beta3 / Q000494:pharmacology*; D020257:Ventricular Remodeling / Q000187:drug effects* / Q000235:genetics
chemical_list: D044403:Receptor, Serotonin, 5-HT2B; D018125:Receptors, Transforming Growth Factor beta; D051785:Smad Proteins; D053782:Transforming Growth Factor beta3; D012701:Serotonin; D017346:Protein Serine-Threonine Kinases; D000077293:Receptor, Transforming Growth Factor-beta Type I