Neural tube defects in embryos of diabetic mice: role of the Pax-3 gene and apoptosis.
Neural tube defects are among the most common of the malformations associated with diabetic embryopathy. To study the molecular mechanisms by which neural tube defects occur during diabetic pregnancy, we have developed a new experimental system using pregnant diabetic mice. In this system, the rate of neural tube defects is about three times higher in embryos of diabetic mice than in embryos of nondiabetic mice. Most of the defects affected presumptive midbrain and hindbrain structures and included open defects (i.e., exencephaly) and gross maldevelopment. By semiquantitative reverse transcription-polymerase chain reaction and in situ hybridization, we found that expression of Pax-3, a gene required for neural tube closure in the area of the midbrain and hindbrain, is significantly reduced in the embryos of diabetic mice. The same regions of the neural tube where Pax-3 had been underexpressed were found subsequently to contain high concentrations of cells undergoing apoptosis. Reduced expression of Pax-3 appears to be responsible for this apoptosis because apoptotic cells were also found at sites of neural tube defects in embryos carrying null mutation of the Pax-3 gene. Finally, mouse strains that carry null mutations in Pax-3 develop neural tube defects that resemble the malformations that occur in embryos of diabetic mice. These results suggest that Pax-3 is an important developmental control gene, expression of which is disturbed in embryos of diabetic mice, and that as a consequence, apoptosis of the neural tube occurs. This pathway may be responsible for many of the neural tube defects resulting from diabetic pregnancy.
Phelan
S A
SA
Section on Molecular Biology, Joslin Diabetes Center, Harvard Medical School, Boston, Massachusetts 02215, USA.
Ito
M
M
Loeken
M R
MR
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000235:genetics*; D003921:Diabetes Mellitus, Experimental / Q000139:chemically induced / Q000150:complications* / Q000196:embryology; D003922:Diabetes Mellitus, Type 1 / Q000139:chemically induced / Q000150:complications* / Q000196:embryology; D004622:Embryo, Mammalian / Q000002:abnormalities* / Q000502:physiology; D005260:Female; D015870:Gene Expression / Q000235:genetics; D017403:In Situ Hybridization; D008098:Litter Size; D051379:Mice; D008813:Mice, Inbred ICR; D009436:Neural Tube Defects / Q000175:diagnosis / Q000209:etiology* / Q000235:genetics; D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011254:Pregnancy in Diabetics / Q000139:chemically induced / Q000150:complications* / Q000196:embryology; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D014157:Transcription Factors
D004268:DNA-Binding Proteins; D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors; D012333:RNA, Messenger; D014157:Transcription Factors; C068686:Pax3 protein, mouse