Folic acid remodels chromatin on Hes1 and Neurog2 promoters during caudal neural tube development.
The mechanism(s) behind folate rescue of neural tube closure are not well understood. In this study we show that maternal intake of folate prior to conception reverses the proliferation potential of neural crest stem cells in homozygous Splotch embryos (Sp(-/-)) via epigenetic mechanisms. It is also shown that the pattern of differentiation seen in these cells is similar to wild-type (WT). Cells from open caudal neural tubes of Sp(-/-) embryos exhibit increased H3K27 methylation and decreased expression of KDM6B possibly due to up-regulation of KDM6B targeting micro-RNAs such as miR-138, miR-148a, miR-185, and miR-339-5p. In our model, folate reversed these epigenetic marks in folate-rescued Sp(-/-) embryos. Using tissue from caudal neural tubes of murine embryos we also examined H3K27me2 and KDM6B association with Hes1 and Neurog2 promoters at embryonic day E10.5, the proliferative stage, and E12.5, when neural differentiation begins. In Sp(-/-) embryos compared with WT, levels of H3K27me2 associated with the Hes1 promoter were increased at E10.5, and levels associated with the Neurog2 promoter were increased at E12.5. KDM6B association with Hes1 and Neurog2 promoters was inversely related to H3K27me2 levels. These epigenetic changes were reversed in folate-rescued Sp(-/-) embryos. Thus, one of the mechanisms by which folate may rescue the Sp(-/-) phenotype is by increasing the expression of KDM6B, which in turn decreases H3K27 methylation marks on Hes1 and Neurog2 promoters thereby affecting gene transcription.
Ichi
Shunsuke
S
Laboratory of Developmental Neurobiology, Department of Pediatric Neurosurgery, Developmental Biology Program, Children's Memorial Research Center and Northwestern University Feinberg School of Medicine, Chicago, Illinois 60614, USA.
Costa
Fabricio F
FF
Bischof
Jared M
JM
Nakazaki
Hiromichi
H
Shen
Yueh-Wei
YW
Boshnjaku
Vanda
V
Sharma
Saurabh
S
Mania-Farnell
Barbara
B
McLone
David G
DG
Tomita
Tadanori
T
Soares
Marcelo B
MB
Mayanil
Chandra S K
CS
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002490:Central Nervous System / Q000196:embryology; D042002:Chromatin Assembly and Disassembly / Q000187:drug effects* / Q000235:genetics; D047369:Chromatin Immunoprecipitation; D004622:Embryo, Mammalian / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D057890:Epigenomics; D005260:Female; D005455:Fluorescent Antibody Technique; D005492:Folic Acid / Q000008:administration & dosage* / Q000494:pharmacology; D006657:Histones / Q000235:genetics / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D007124:Immunoenzyme Techniques; D047468:Immunoprecipitation; D056484:Jumonji Domain-Containing Histone Demethylases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D008156:Luciferases / Q000378:metabolism; D008297:Male; D008745:Methylation; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000502:physiology; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000378:metabolism; D058953:Neural Stem Cells / Q000187:drug effects / Q000378:metabolism; D054259:Neural Tube / Q000187:drug effects* / Q000196:embryology*; D009436:Neural Tube Defects / Q000517:prevention & control; D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors / Q000502:physiology; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000072056:Transcription Factor HES-1; D014803:Vitamin B Complex / Q000008:administration & dosage / Q000494:pharmacology
D051792:Basic Helix-Loop-Helix Proteins; D005492:Folic Acid; D006657:Histones; D018398:Homeodomain Proteins; D056484:Jumonji Domain-Containing Histone Demethylases; D008156:Luciferases; D035683:MicroRNAs; D009419:Nerve Tissue Proteins; D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D000072056:Transcription Factor HES-1; D014803:Vitamin B Complex; C489734:Hes1 protein, mouse; C102938:Neurog2 protein, mouse; C068686:Pax3 protein, mouse; C523761:Kdm6b protein, mouse