title: Early migratory rat neural crest cells express functional gap junctions: evidence that neural crest cell survival requires gap junction function.
abstract: Gap junctions mediate crucial intercellular interactions during development. This study provides evidence that early migrating rat neural crest cells assemble functional gap junctions, as demonstrated by dye transfer following microinjection of single cells, which were phenotypically identified as neural crest cells by their expression of the low- affinity nerve growth factor receptor. An immunohistochemical analysis using connexin- specific antibodies revealed that migrating rat neural crest cells express the gap junction constituents connexins 43 (Cx 43) and Cx 46. We tested the hypothesis that gap junctions play an important role during early neural crest cell development by perturbing their function in migrating neural crest cells. Our data show that markedly decreasing gap junction communication between these neural crest cells in vitro with either 18alpha-glycyrrhetinic acid or anandamide decreases their survival, whereas oleamide, a less effective blocker of connexon function, had quantitatively less effect on neural crest cell death. This cell death was associated with the occurrence of DNA nicking as detected by the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick end-labelling (TUNEL) procedure, suggesting cell death via apoptosis. The effect of 18alpha-glycyrrhetinic acid and anandamide on neural crest cell survival was reversible and was not mimicked by the structurally related compounds glycyrrhizic acid and palmitoylethanolamide, respectively, which do not uncouple cells. These results indicate that gap junctions are necessary for the survival of spinal neural crest cells.
authors.affiliation: Neurology Research, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA.
authors.forename: P
authors.initials: P
authors.lastname: Bannerman
authors.forename: W
authors.initials: W
authors.lastname: Nichols
authors.forename: S
authors.initials: S
authors.lastname: Puhalla
authors.forename: T
authors.initials: T
authors.lastname: Oliver
authors.forename: M
authors.initials: M
authors.lastname: Berman
authors.forename: D
authors.initials: D
authors.lastname: Pleasure
mesh_terms: D020003:1-Octanol / Q000494:pharmacology; D000367:Age Factors; D000818:Animals; D000911:Antibodies, Monoclonal; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D001095:Arachidonic Acids / Q000494:pharmacology; D001710:Biotin / Q000031:analogs & derivatives; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002465:Cell Movement / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D018678:Cholinergic Agents; D018031:Connexin 43 / Q000032:analysis; D017630:Connexins / Q000032:analysis; D063388:Endocannabinoids; D005260:Female; D017629:Gap Junctions / Q000737:chemistry / Q000502:physiology*; D017930:Genes, Reporter; D006034:Glycyrrhetinic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D006993:Hypnotics and Sedatives / Q000494:pharmacology; D000922:Immunotoxins; D009699:N-Glycosyl Hydrolases; D009432:Neural Crest / Q000166:cytology*; D009474:Neurons / Q000737:chemistry / Q000166:cytology*; D009829:Oleic Acids / Q000494:pharmacology; D053284:Polyunsaturated Alkamides; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D020800:Receptor, Nerve Growth Factor / Q000235:genetics; D054789:Ribosome Inactivating Proteins, Type 1; D000076985:Saporins; D014475:Uncoupling Agents / Q000494:pharmacology
chemical_list: C085696:192 IgG-saporin; D000911:Antibodies, Monoclonal; D001095:Arachidonic Acids; D002121:Calcium Channel Blockers; D018678:Cholinergic Agents; D018031:Connexin 43; D017630:Connexins; D063388:Endocannabinoids; D006993:Hypnotics and Sedatives; D000922:Immunotoxins; D009829:Oleic Acids; D053284:Polyunsaturated Alkamides; D020800:Receptor, Nerve Growth Factor; D054789:Ribosome Inactivating Proteins, Type 1; D014475:Uncoupling Agents; C082917:GJA3 protein, human; C070614:neurobiotin; C119129:18alpha-glycyrrhetinic acid; D001710:Biotin; C029407:oleylamide; D009699:N-Glycosyl Hydrolases; D000076985:Saporins; D020003:1-Octanol; D006034:Glycyrrhetinic Acid; C078814:anandamide