title: Control of the collective migration of enteric neural crest cells by the Complement anaphylatoxin C3a and N-cadherin.
abstract: We analyzed the cellular and molecular mechanisms governing the adhesive and migratory behavior of enteric neural crest cells (ENCCs) during their collective migration within the developing mouse gut. We aimed to decipher the role of the complement anaphylatoxin C3a during this process, because this well-known immune system attractant has been implicated in cephalic NCC co-attraction, a process controlling directional migration. We used the conditional Ht-PA-cre transgenic mouse model allowing a specific ablation of the N-cadherin gene and the expression of a fluorescent reporter in migratory ENCCs without affecting the central nervous system. We performed time-lapse videomicroscopy of ENCCs from control and N-cadherin mutant gut explants cultured on fibronectin (FN) and micropatterned FN-stripes with C3a or C3aR antagonist, and studied cell migration behavior with the use of triangulation analysis to quantify cell dispersion. We performed ex vivo gut cultures with or without C3aR antagonist to determine the effect on ENCC behavior. Confocal microscopy was used to analyze the cell-matrix adhesion properties. We provide the first demonstration of the localization of the complement anaphylatoxin C3a and its receptor on ENCCs during their migration in the embryonic gut. C3aR receptor inhibition alters ENCC adhesion and migration, perturbing directionality and increasing cell dispersion both in vitro and ex vivo. N-cadherin-null ENCCs do not respond to C3a co-attraction. These findings indicate that C3a regulates cell migration in a N-cadherin-dependent process. Our results shed light on the role of C3a in regulating collective and directional cell migration, and in ganglia network organization during enteric nervous system ontogenesis. The detection of an immune system chemokine in ENCCs during ENS development may also shed light on new mechanisms for gastrointestinal disorders.
authors.affiliation: Institut Curie/CNRS UMR144, Paris, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Broders-Bondon
authors.affiliation: Cell and Tissue Imaging Facility, PICT-IBiSA, Paris, France.
authors.forename: Perrine
authors.initials: P
authors.lastname: Paul-Gilloteaux
authors.affiliation: Institut Curie/CNRS UMR144, Paris, France; Université Paris Est, Faculté de médecine, Créteil 94000, France; INSERM, U955, Team 6, Créteil, 94000, France.
authors.forename: Elodie
authors.initials: E
authors.lastname: Gazquez
authors.affiliation: Institut Curie/CNRS UMR144, Paris, France.
authors.forename: Julie
authors.initials: J
authors.lastname: Heysch
authors.affiliation: Institut Curie/CNRS UMR144, Paris, France.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Piel
authors.affiliation: Department of Cell and Developmental Biology, University College, London, UK.
authors.forename: Roberto
authors.initials: R
authors.lastname: Mayor
authors.affiliation: University of Pennsylvannia, Philadelphia, USA.
authors.forename: John D
authors.initials: JD
authors.lastname: Lambris
authors.affiliation: Institut Curie/CNRS UMR144, Paris, France; Université Paris Est, Faculté de médecine, Créteil 94000, France; INSERM, U955, Team 6, Créteil, 94000, France. Electronic address: sylvie.dufour@inserm.fr.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Dufour
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D015820:Cadherins / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D002448:Cell Adhesion; D002465:Cell Movement; D015926:Complement C3a / Q000819:agonists / Q000502:physiology*; D003433:Crosses, Genetic; D017615:Enteric Nervous System / Q000166:cytology / Q000196:embryology*; D005109:Extracellular Matrix / Q000502:physiology; D005260:Female; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D008297:Male; D051379:Mice; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D009432:Neural Crest / Q000166:cytology*; D043562:Receptors, G-Protein-Coupled / Q000037:antagonists & inhibitors / Q000502:physiology; D059008:Time-Lapse Imaging
chemical_list: D001426:Bacterial Proteins; C000609604:C3a-derived anaphylatoxin receptor, mouse; D015820:Cadherins; C489083:Cdh2 protein, mouse; D008164:Luminescent Proteins; D043562:Receptors, G-Protein-Coupled; C054562:yellow fluorescent protein, Bacteria; D015926:Complement C3a
keywords: Adhesion; Co-attraction; Complement anaphylatoxin C3a; Enteric nervous system; Migration; N-cadherin; Neural crest cells