Functional interactions between the LRP6 WNT co-receptor and folate supplementation.
Crooked tail (Cd) mice bear a gain-of-function mutation in Lrp6, a co-receptor for canonical WNT signaling, and are a model of neural tube defects (NTDs), preventable with dietary folic acid (FA) supplementation. Whether the FA response reflects a direct influence of FA on LRP6 function was tested with prenatal supplementation in LRP6-deficient embryos. The enriched FA (10 ppm) diet reduced the occurrence of birth defects among all litters compared with the control (2 ppm FA) diet, but did so by increasing early lethality of Lrp6(-/-) embryos while actually increasing NTDs among nulls alive at embryonic days 10-13 (E10-13). Proliferation in cranial neural folds was reduced in homozygous Lrp6(-/-) mutants versus wild-type embryos at E10, and FA supplementation increased proliferation in wild-type but not mutant neuroepithelia. Canonical WNT activity was reduced in LRP6-deficient midbrain-hindbrain at E9.5, demonstrated in vivo by a TCF/LEF-reporter transgene. FA levels in media modulated the canonical WNT response in NIH3T3 cells, suggesting that although FA was required for optimal WNT signaling, even modest FA elevations attenuated LRP5/6-dependent canonical WNT responses. Gene expression analysis in embryos and adults showed striking interactions between targeted Lrp6 deficiency and FA supplementation, especially for mitochondrial function, folate and methionine metabolism, WNT signaling and cytoskeletal regulation that together implicate relevant signaling and metabolic pathways supporting cell proliferation, morphology and differentiation. We propose that FA supplementation rescues Lrp6(Cd/Cd) fetuses by normalizing hyperactive WNT activity, whereas in LRP6-deficient embryos, added FA further attenuates reduced WNT activity, thereby compromising development.
Gray
Jason D
JD
Laboratory of Neurogenetics and Development, Graduate Program in Neuroscience, Weill Medical College of Cornell University, 1300 York Avenue, New York, NY 10021, USA.
Nakouzi
Ghunwa
G
Slowinska-Castaldo
Bozena
B
Dazard
Jean-Eudes
JE
Rao
J Sunil
JS
Nadeau
Joseph H
JH
Ross
M Elizabeth
ME
D000818:Animals; D019587:Dietary Supplements; D004195:Disease Models, Animal; D005492:Folic Acid / Q000008:administration & dosage* / Q000378:metabolism / Q000494:pharmacology; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D007150:Immunohistochemistry; D026502:LDL-Receptor Related Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D060488:Low Density Lipoprotein Receptor-Related Protein-6; D008715:Methionine / Q000378:metabolism; D051379:Mice; D008809:Mice, Inbred C3H; D008928:Mitochondria / Q000378:metabolism; D009154:Mutation; D041681:NIH 3T3 Cells; D009432:Neural Crest / Q000002:abnormalities / Q000187:drug effects / Q000254:growth & development; D009436:Neural Tube Defects / Q000196:embryology / Q000235:genetics / Q000378:metabolism / Q000517:prevention & control; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D017422:Sequence Analysis, DNA; D015398:Signal Transduction / Q000187:drug effects; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism*
D026502:LDL-Receptor Related Proteins; D060488:Low Density Lipoprotein Receptor-Related Protein-6; C555476:Lrp6 protein, mouse; D012333:RNA, Messenger; D051153:Wnt Proteins; D005492:Folic Acid; D008715:Methionine