title: GDNF and endothelin 3 regulate migration of enteric neural crest-derived cells via protein kinase A and Rac1.
abstract: Enteric neural crest-derived cells (ENCCs) migrate from the anterior foregut in a rostrocaudal direction to colonize the entire gastrointestinal tract and to form the enteric nervous system. Genetic approaches have identified many signaling molecules regulating the migration of ENCCs; however, it remains elusive how the activities of the signaling molecules are regulated spatiotemporally during migration. In this study, transgenic mice expressing biosensors based on Förster resonance energy transfer were generated to video the activity changes of the signaling molecules in migrating ENCCs. In an organ culture of embryonic day 11.25 (E11.25) to E13 guts, ENCCs at the rostral wavefront migrated as a cellular chain faster than the following ENCCs that formed a network. The faster-migrating cells at the wavefront exhibited lower protein kinase A (PKA) activity than did the slower-migrating trailing cells. The activities of Rac1 and Cdc42 exhibited an inverse correlation with the PKA activity, and PKA activation decreased the Rac1 activity and migration velocity. PKA activity in ENCCs was correlated positively with the distribution of GDNF and inversely with the distribution of endothelin 3 (ET-3). Accordingly, PKA was activated by GDNF and inhibited by ET-3 in cultured ENCCs. Finally, although the JNK and ERK pathways were previously reported to control the migration of ENCCs, we did not find any correlation of JNK or ERK activity with the migration velocities. These results suggest that external cues regulate the migration of ENCCs by controlling PKA activity, but not ERK or JNK activity, and argue for the importance of live imaging of signaling molecule activities in developing organs.
authors.affiliation: Laboratory of Bioimaging and Cell Signaling, Graduate School of Biostudies, Graduate School of Medicine, Kyoto University, Kyoto 606-8315, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Goto
authors.forename: Kenta
authors.initials: K
authors.lastname: Sumiyama
authors.forename: Yuji
authors.initials: Y
authors.lastname: Kamioka
authors.forename: Eiji
authors.initials: E
authors.lastname: Nakasyo
authors.forename: Keisuke
authors.initials: K
authors.lastname: Ito
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Iwasaki
authors.forename: Hideki
authors.initials: H
authors.lastname: Enomoto
authors.forename: Michiyuki
authors.initials: M
authors.lastname: Matsuda
mesh_terms: D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000367:Age Factors; D000818:Animals; D015374:Biosensing Techniques; D050882:CREB-Binding Protein / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004064:Digestive System / Q000166:cytology / Q000196:embryology; D004622:Embryo, Mammalian; D019334:Endothelin-3 / Q000378:metabolism* / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D031541:Fluorescence Resonance Energy Transfer; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000378:metabolism* / Q000494:pharmacology; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000187:drug effects / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D009432:Neural Crest / Q000166:cytology*; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D009924:Organ Culture Techniques; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D011247:Pregnancy; D013873:Thionucleotides / Q000494:pharmacology; D020764:cdc42 GTP-Binding Protein / Q000235:genetics / Q000378:metabolism; D020830:rac1 GTP-Binding Protein / Q000378:metabolism*; D000097573:Red Fluorescent Protein
chemical_list: D015124:8-Bromo Cyclic Adenosine Monophosphate; D050882:CREB-Binding Protein; D017868:Cyclic AMP-Dependent Protein Kinases; D019334:Endothelin-3; D004791:Enzyme Inhibitors; D051100:Glial Cell Line-Derived Neurotrophic Factor; D008164:Luminescent Proteins; D019869:Phosphatidylinositol 3-Kinases; D013873:Thionucleotides; D020764:cdc42 GTP-Binding Protein; D020830:rac1 GTP-Binding Protein; D000097573:Red Fluorescent Protein; C105593:8-bromoadenosine-3',5'-cyclic monophosphorothioate