title: Novel migrating mouse neural crest cell assay system utilizing P0-Cre/EGFP fluorescent time-lapse imaging.
abstract: BACKGROUND
  Neural crest cells (NCCs) are embryonic, multipotent stem cells. Their long-range and precision-guided migration is one of their most striking characteristics. We previously reported that P0-Cre/CAG-CAT-lacZ double-transgenic mice showed significant lacZ expression in tissues derived from NCCs.
  
  
  RESULTS
  In this study, by embedding a P0-Cre/CAG-CAT-EGFP embryo at E9.5 in collagen gel inside a culture glass slide, we were able to keep the embryo developing ex vivo for more than 24 hours; this development was with enough NCC fluorescent signal intensity to enable single-cell resolution analysis, with the accompanying NCC migration potential intact and with the appropriate NCC response to the extracellular signal maintained. By implantation of beads with absorbed platelet-derived growth factor-AA (PDGF-AA), we demonstrated that PDGF-AA acts as an NCC-attractant in embryos.We also performed assays with NCCs isolated from P0-Cre/CAG-CAT-EGFP embryos on culture plates. The neuromediator 5-hydroxytryptamine (5-HT) has been known to regulate NCC migration. We newly demonstrated that dopamine, in addition to 5-HT, stimulated NCC migration in vitro. Two NCC populations, with different axial levels of origins, showed unique distribution patterns regarding migration velocity and different dose-response patterns to both 5-HT and dopamine.
  
  
  CONCLUSIONS
  Although avian species predominated over the other species in the NCC study, our novel system should enable us to use mice to assay many different aspects of NCCs in embryos or on culture plates, such as migration, division, differentiation, and apoptosis.
authors: Division of Developmental Genetics, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto-City, Kumamoto, 860-0811, Japan. mkawa@gpo.kumamoto-u.ac.jp
authors: Minoru
authors: M
authors: Kawakami
authors: Masafumi
authors: M
authors: Umeda
authors: Naomi
authors: N
authors: Nakagata
authors: Toru
authors: T
authors: Takeo
authors: Ken-Ichi
authors: K
authors: Yamamura
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002465:Cell Movement; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009432:Neural Crest / Q000166:cytology* / Q000378:metabolism; D010982:Platelet-Derived Growth Factor / Q000378:metabolism; D012701:Serotonin / Q000378:metabolism; D059008:Time-Lapse Imaging / Q000379:methods*
chemical_list: D010982:Platelet-Derived Growth Factor; C485184:enhanced green fluorescent protein; C075278:platelet-derived growth factor A; D049452:Green Fluorescent Proteins; D012701:Serotonin