title: Altered gene expression in rat cranial neural crest cells exposed to a teratogenic glucose concentration in vitro: paradoxical downregulation of antioxidative defense genes.
abstract: BACKGROUND
  Diabetic pregnancy is associated with increased risk of malformation in the infant. Diabetes-induced anomalies of the face and heart are strongly correlated with neural crest cell (NCC) maldevelopment. We aimed to study glucose-induced alterations of mRNA levels in cranial and trunk NCCs isolated from rat embryos with increased risk of developing mandibular and cardiac malformations in diabetic pregnancy.
  
  
  METHODS
  Inbred Sprague-Dawley rat embryos were used for NCC isolation from neural tube explants. The migrating cells were exposed to 5.5 or 30 mmol/l glucose concentration for 48 hr, harvested, and prepared for gene expression measurement by RT-PCR or immunostaining with either distal-less (Dlx) or AP-2-α antibodies.
  
  
  RESULTS
  Evaluation of the immunostained slides showed that approximately 75% of the cells were of NCC origin. Exposure to 30 mM glucose decreased mRNA levels of Copper-Zinc superoxide dismutase, manganese superoxide dismutase, extracellular superoxide dismutase, Catalase, Gpx-1, Nrf2, poly-ADP ribose polymerase, B-cell leukemia/lymphoma protein 2, and β-Catenin genes in cranial neural crest explant cultures. In addition, Pax-3, Pax-6, Wnt3a, and Apc mRNA levels were decreased by high glucose exposure in both cranial and trunk neural crest explant cultures.
  
  
  CONCLUSIONS
  Cranial NCCs diminish their mRNA levels of antioxidative enzymes and the Nrf2 response factor, as well as the antiapoptotic B-cell leukemia/lymphoma protein 2 gene, in response to increased ambient glucose concentration. Furthermore, both cranial and trunk NCC decrease the mRNA levels of the transcription factors Pax-3 and Pax-6, as well as key components of the Wnt pathway. These patterns of glucose-altered gene expression in a developmentally important cell population may be of etiological importance for NCC-associated malformations in diabetic pregnancy.
authors.affiliation: Department of Medical Cell Biology, Uppsala University, Biomedical Center, Sweden. parri.wentzel@mcb.uu.se
authors.forename: Parri
authors.initials: P
authors.lastname: Wentzel
authors.forename: Ulf J
authors.initials: UJ
authors.lastname: Eriksson
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D048909:Diabetes Complications / Q000235:genetics; D004622:Embryo, Mammalian / Q000187:drug effects; D005136:Eye Proteins / Q000096:biosynthesis; D005260:Female; D015870:Gene Expression / Q000187:drug effects*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D005947:Glucose / Q000008:administration & dosage / Q000633:toxicity*; D018398:Homeodomain Proteins / Q000096:biosynthesis; D009432:Neural Crest / Q000166:cytology / Q000187:drug effects* / Q000196:embryology / Q000378:metabolism; D000072041:PAX3 Transcription Factor; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000096:biosynthesis; D011247:Pregnancy; D011254:Pregnancy in Diabetics / Q000503:physiopathology*; D011518:Proto-Oncogene Proteins / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012097:Repressor Proteins / Q000096:biosynthesis; D013723:Teratogens / Q000633:toxicity*; D060449:Wnt Signaling Pathway / Q000187:drug effects
chemical_list: D005136:Eye Proteins; D018398:Homeodomain Proteins; D000072041:PAX3 Transcription Factor; C530820:PAX3 protein, rat; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; C000605733:Pax6 protein, rat; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; D013723:Teratogens; D005947:Glucose