title: Fetal neural tube stem cells from Pax3 mutant mice proliferate, differentiate, and form synaptic connections when stimulated with folic acid.
abstract: Although maternal intake of folic acid (FA) prevents neural tube defects in 70% of the population, the exact mechanism of prevention has not been elucidated. We hypothesized that FA affects neural stem cell (NSC) proliferation and differentiation. This hypothesis was examined in a folate-responsive spina bifida mouse model, Splotch (Sp(-/-)), which has a homozygous loss-of-function mutation in the Pax3 gene. Neurospheres were generated with NSCs from the lower lumbar neural tube of E10.5 wild-type (WT) and Sp(-/-) embryos, in the presence and absence of FA. In the absence of FA, the number of neurospheres generated from Sp(-/-) embryos compared with WT was minimal (P<0.05). Addition of FA to Sp(-/-) cultures increased the expression of a Pax3 downstream target, fgfr4, and rescued NSC proliferative potential, as demonstrated by a significant increase in neurosphere formation (P<0.01). To ascertain if FA affected cell differentiation, FA-stimulated Sp(-/-) neurospheres were allowed to differentiate in the continued presence or absence of FA. Neurospheres from both conditions expressed multi-potent stem cell characteristics and the same differentiation potential as WT. Further, multiple neurospheres from both WT and FA-stimulated Sp(-/-) cell cultures formed extensive synaptic connections. On the whole, FA-mediated rescue of neural tube defects in Sp(-/-) embryos promotes NSC proliferation at an early embryonic stage. FA-stimulated Sp(-/-) neurospheres differentiate and form synaptic connections, comparable to WT.
authors.affiliation: Developmental Biology Program, Division of Pediatric Neurosurgery, Children's Memorial Hospital and Research Center, Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60614, USA.
authors.forename: Shunsuke
authors.initials: S
authors.lastname: Ichi
authors.forename: Hiromichi
authors.initials: H
authors.lastname: Nakazaki
authors.forename: Vanda
authors.initials: V
authors.lastname: Boshnjaku
authors.forename: Ravneet Monny
authors.initials: RM
authors.lastname: Singh
authors.forename: Barbara
authors.initials: B
authors.lastname: Mania-Farnell
authors.forename: Guifa
authors.initials: G
authors.lastname: Xi
authors.forename: David G
authors.initials: DG
authors.lastname: McLone
authors.forename: Tadanori
authors.initials: T
authors.lastname: Tomita
authors.forename: Chandra Shekhar K
authors.initials: CS
authors.lastname: Mayanil
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004195:Disease Models, Animal; D004622:Embryo, Mammalian; D053686:Fetal Stem Cells / Q000187:drug effects* / Q000473:pathology; D005333:Fetus; D005492:Folic Acid / Q000494:pharmacology*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D055786:Gene Knockout Techniques; D006720:Homozygote; D051379:Mice; D018345:Mice, Knockout; D058953:Neural Stem Cells / Q000187:drug effects* / Q000473:pathology; D054259:Neural Tube / Q000187:drug effects* / Q000196:embryology / Q000473:pathology; D009474:Neurons / Q000187:drug effects / Q000473:pathology; D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors / Q000172:deficiency / Q000235:genetics; D051499:Receptor, Fibroblast Growth Factor, Type 4 / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D016135:Spinal Dysraphism / Q000188:drug therapy* / Q000196:embryology / Q000235:genetics / Q000473:pathology; D013569:Synapses / Q000187:drug effects / Q000502:physiology
chemical_list: D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors; C068686:Pax3 protein, mouse; D005492:Folic Acid; C496372:Fgfr4 protein, mouse; D051499:Receptor, Fibroblast Growth Factor, Type 4