High-affinity folate receptor in cardiac neural crest migration: a gene knockdown model using siRNA.
Folate supplementation reduces the incidence of congenital heart defects, but the nature of this protective mechanism remains unclear. Immunolabeling demonstrated that the neural tube and neural crest (NC) cells were rich in the high-affinity folate receptor FOLR1and during the early stages of development FOLR1 was found principally in these cells. Suppression of Folr1 expression in the nascent cardiac NC by site-directed short-interfering RNA (siRNA) altered cardiac NC cell mitosis and subsequent migration patterns leading to abnormal development of the pharyngeal arch arteries (PAA) and outflow tract. qPCR analysis demonstrated that the siRNA treatment significantly reduced Folr1 24 hr after treatment. These treatments also significantly reduced mitosis in the neural tube, but adjacent, nontreated areas were unaffected. In summary, a brief reduction in the expression of Folr1 during a critical stage of NC development had long-term consequences for the development of the PAA and outflow tract.
Department of Genetics, Cell Biology and Anatomy, University of Nebraska Medical Center, Omaha, Nebraska 68198-5805, USA.
Thomas H
TH
Rosenquist
Tammy
T
Chaudoin
Richard H
RH
Finnell
Gregory D
GD
Bennett
D000818:Animals; D030801:Animals, Genetically Modified; D001158:Arteries / Q000187:drug effects / Q000196:embryology / Q000378:metabolism; D001934:Branchial Region / Q000098:blood supply / Q000187:drug effects / Q000196:embryology / Q000378:metabolism; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002465:Cell Movement / Q000187:drug effects / Q000235:genetics*; D002642:Chick Embryo; D058973:Folate Receptors, GPI-Anchored; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D055785:Gene Knockdown Techniques; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006321:Heart / Q000187:drug effects / Q000196:embryology*; D023421:Models, Animal; D009432:Neural Crest / Q000187:drug effects / Q000196:embryology / Q000378:metabolism* / Q000502:physiology; D034741:RNA, Small Interfering / Q000494:pharmacology*; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D013379:Substrate Specificity; D013997:Time Factors
D002352:Carrier Proteins; D058973:Folate Receptors, GPI-Anchored; D034741:RNA, Small Interfering; D011956:Receptors, Cell Surface; D049452:Green Fluorescent Proteins