PBX, MEIS, and IGF-I are potential mediators of retinoic acid-induced proximodistal limb reduction defects
| Autor: | Kenneth J. Soprano, Pu Qin, Rebecca Cimildoro, Dianne Robert Soprano, D. M. Kochhar |
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| Rok vydání: | 2002 |
| Předmět: |
Gene isoform
Embryology animal structures Microarray Ectromelia Health Toxicology and Mutagenesis Limb Deformities Congenital Retinoic acid Mice Inbred Strains Tretinoin Biology Toxicology Mice chemistry.chemical_compound Western blot Pregnancy Gene expression medicine Animals Protein Isoforms Limb development RNA Messenger Insulin-Like Growth Factor I Maternal-Fetal Exchange Oligonucleotide Array Sequence Analysis Homeodomain Proteins Genetics medicine.diagnostic_test Microarray analysis techniques Days post coitum Abnormalities Drug-Induced Gene Expression Regulation Developmental Molecular biology Teratogens chemistry Female Transcription Factors Developmental Biology |
| Zdroj: | Teratology. 66:224-234 |
| ISSN: | 1096-9926 0040-3709 |
| DOI: | 10.1002/tera.10082 |
| Popis: | Background Phocomelia, which is primarily due to a disruption in the proximodistal axis, is found in virtually all mouse embryos exposed to high doses of retinoic acid (RA) on 11 days post coitum (dpc). Methods To identify genes that potentially mediate the effects of retinoic acid (RA) on limb development, we have examined the expression of 9,000 clones from the IMAGE consortium by microarray analysis of RNA isolated from 11 dpc mouse forelimbs exposed to RA or vehicle for 6 hr. Eight genes that demonstrated altered expression were chosen for further study of their mRNA levels using RT-PCR. Protein levels were determined by Western blot analysis. Results Of the 9,000 genes examined in the microarray, approximately 111 demonstrated altered expression (33 known genes and 78 ESTs). Of the eight known genes chosen for further study using RT-PCR, four mRNAs (PBX1a, PBX1b, IGF-Ia, and IGF-Ib) demonstrated consistent elevation (≈3-fold) in their levels after RA treatment in both the forelimbs and hindlimbs as early as 3 hr after RA treatment. In addition to the two PBX1 isoforms, the mRNA level of the other two subtypes (PBX2 and PBX3) and the level of PBX1/2/3 protein were also found to be elevated in limb buds after RA treatment. Finally, we examined the expression of MEIS1, MEIS2, and MEIS3 because these proteins are necessary for PBX nuclear localization. The mRNA level of all three subtypes of MEIS were elevated approximately three- to four-fold in both the forelimbs and hindlimbs after RA treatment. Conclusions Because both PBX and MEIS (and their orthologs) are believed to be involved in the control of proximodistal axis formation in mouse and fly limbs and IGFs in the development of limbs, we suggest that increases in PBX, MEIS and IGF-1 mRNA levels may contribute to proximodistal limb reduction defects caused by teratogenic doses of RA. Teratology 66:224–234, 2002. © 2002 Wiley-Liss, Inc. |
| Databáze: | OpenAIRE |
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