Functional BMP receptor in endocardial cells is required in atrioventricular cushion mesenchymal cell formation in chick.
Transformation of atrioventricular (AV) canal endocardium into invasive mesenchyme correlates spatially and temporally with the expression of bone morphogenetic protein (BMP)-2 in the AV myocardium. We revealed the presence of mRNA of Type I BMP receptors, BMPR-1A (ALK3), BMPR-1B (ALK6) and ALK2 in chick AV endocardium at stage-14(-), the onset of epithelial to mesenchymal transformation (EMT), by RT-PCR and localized BMPR-1B mRNA in the endocardium by in situ hybridization. To circumvent the functional redundancies among the Type I BMP receptors, we applied dominant-negative (dn) BMPR-1B-viruses to chick AV explants and whole-chick embryo cultures to specifically block BMP signaling in AV endocardium during EMT. dnBMPR-1B-virus infection of AV endocardial cells abolished BMP-2-supported AV endocardial EMT. Conversely, caBMPR-1B-virus infection promoted AV endocardial EMT in the absence of AV myocardium. Moreover, dnBMPR-1B-virus treatments significantly reduced myocardially supported EMT in AV endocardial-myocardial co-culture. AV cushion mesenchymal cell markers, alpha-smooth muscle actin (SMA), and TGFbeta3 in the endocardial cells were promoted by caBMPR-1B and reduced by dnBMPR-1B infection. Microinjection of the virus into the cardiac jelly in the AV canal at stage-13 in vivo (ovo) revealed that the dnBMPR-1B-virus-infected cells remained in the endocardial epithelium, whereas caBMPR-1B-infected cells invaded deep into the cushions. These results provide evidence that BMP signaling through the AV endocardium is required for the EMT and the activation of the BMP receptor in the endocardium can promote AV EMT in the chick.
Okagawa
Hiroto
H
Department of Cell Biology and Anatomy and Cardiovascular Developmental Biology Center, Medical University of South Carolina, 171 Ashley Avenue, Charleston, SC 29425, USA. jefata@lbl.gov <jefata@lbl.gov>
Markwald
Roger R
RR
Sugi
Yukiko
Y
D000818:Animals; D052005:Bone Morphogenetic Protein Receptors, Type I / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism; D002642:Chick Embryo; D018920:Coculture Techniques; D003094:Collagen / Q000737:chemistry; D004699:Endocardium / Q000166:cytology / Q000196:embryology* / Q000378:metabolism*; D005782:Gels / Q000737:chemistry; D006321:Heart / Q000196:embryology*; D006325:Heart Atria / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D006352:Heart Ventricles / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D008648:Mesoderm / Q000166:cytology* / Q000378:metabolism; D008845:Microinjections; D009206:Myocardium / Q000166:cytology / Q000378:metabolism; D015398:Signal Transduction; D053782:Transforming Growth Factor beta3 / Q000378:metabolism; D000099176:Noggin Protein
D052005:Bone Morphogenetic Protein Receptors, Type I; D002352:Carrier Proteins; D003094:Collagen; D005782:Gels; D053782:Transforming Growth Factor beta3; D000099176:Noggin Protein