A critical role for calponin 2 in vascular development.
Calponin 2 (h2 calponin, CNN2) is an actin-binding protein implicated in cytoskeletal organization. We have found that the expression of calponin 2 is relatively restricted to vasculature from 16 to 30 h post-fertilization during zebrafish (Danio rerio) development. Forty-eight hours after injecting antisense morpholino oligos against calponin 2 into embryos at the 1-4-cell stage, zebrafish demonstrated various cardiovascular defects, including sluggish axial and head circulation, absence of circulation in intersegmental vessels and in the dorsal longitudinal anastomotic vessel, enlarged cerebral ventricles, and pericardial edema, in addition to an excess bending, spiraling tail and twisting of the caudal fin. Knockdown of calponin 2 in the Tg(fli1:EGFP)(y1) zebrafish line (in which a fli1 promoter drives vascular-specific enhanced green fluorescent protein expression) indicated that diminished calponin 2 expression blocked the proper migration of endothelial cells during formation of intersegmental vessels. In vitro studies showed that basic fibroblast growth factor-induced human umbilical vein endothelial cell migration was down-regulated by knockdown of calponin 2 expression using an antisense adenovirus, and overexpression of calponin 2 enhanced migration and hastened wound healing. These events were correlated with activation of mitogen-activated protein kinase; moreover, inhibition of this pathway blocked the promigratory effect of calponin 2. Collectively, these data suggest that calponin 2 plays an important role in the migration of endothelial cells both in vivo and in vitro and that its expression is critical for proper vascular development.
Tang
Jian
J
Renal Division and Center for Study of the Tumor Microenvironment, Department of Medicine, Beth Israel Deaconess Medical Center, Boston, Massachusetts 02215, USA.
Hu
Guang
G
Hanai
Jun-ichi
J
Yadlapalli
Ganesh
G
Lin
Yanfeng
Y
Zhang
Bo
B
Galloway
Jenna
J
Bahary
Nathan
N
Sinha
Sonia
S
Thisse
Bernard
B
Thisse
Christine
C
Jin
Jian-Ping
JP
Zon
Leonard I
LI
Sukhatme
Vikas P
VP
D000595:Amino Acid Sequence; D000818:Animals; D002135:Calcium-Binding Proteins / Q000502:physiology*; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology; D003001:Cloning, Molecular; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D006801:Humans; D020930:MAP Kinase Kinase Kinases / Q000378:metabolism; D008840:Microfilament Proteins / Q000502:physiology*; D008969:Molecular Sequence Data; D018919:Neovascularization, Physiologic / Q000235:genetics / Q000502:physiology*; D016376:Oligonucleotides, Antisense; D010766:Phosphorylation; D015398:Signal Transduction / Q000502:physiology; D015027:Zebrafish / Q000196:embryology / Q000235:genetics; D029961:Zebrafish Proteins / Q000502:physiology*; D000096985:Calponins
D002135:Calcium-Binding Proteins; D048049:Extracellular Signal-Regulated MAP Kinases; D020930:MAP Kinase Kinase Kinases; D008840:Microfilament Proteins; D016376:Oligonucleotides, Antisense; D029961:Zebrafish Proteins; C510190:Cnn2 protein, zebrafish; D000096985:Calponins