Antisense modulation of the coding or regulatory sequence of the folate receptor (Folate binding protein-1) in mouse embryos leads to neural tube defects

被引:14
作者
Hansen, DK [1 ]
Streck, RD
Antony, AC
机构
[1] Natl Ctr Toxicol Res, Div Genet & Reprod Toxicol, Food & Drug Adm, Jefferson, AR 72079 USA
[2] Indiana Univ, Sch Med, Dept Med, Indianapolis, IN USA
[3] Roudebush Vet Affairs Med Ctr, Indianapolis, IN USA
关键词
folic acid; folate receptor; neural tube defects; mouse; rodent whole embryo culture; gene expression; antisense;
D O I
10.1002/bdra.10072
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
BACKGROUND: Although folic acid decreases the incidence of neural tube defects (NTDs) in humans, the mechanism for this protection is unknown. We have employed antisense technology to alter expression of the gene for the folate receptor (folate binding protein-1 [Folbp1]) in mouse embryos cultured in vitro. METHODS: Embryos were explanted on day 8 of gestation and cultured for 44 hr. Several oligodeoxyribonucleotides designed to modulate the coding region or a regulatory sequence in the 5'-untranslated region of Folbp1 were microinjected into the amniotic sac of embryos at the beginning of the culture period. RESULTS: Two different antisense sequences to the 5' and 3' coding region in Folbp1 produced concentration-dependent increases in the number of embryos with NTDs. Coinjection of 5-methyltetrahydrofolate with these sequences decreased the frequency of abnormal embryos. A semi-quantitative RT-PCR technique used to measure the amount of Folbp1 mRNA in treated and control embryos confirmed that the mRNA level was decreased by treatment with the antisense sequences. An antisense oligodeoxyribonucleotide to a 17 base cis regulatory element also generated a concentration-dependent increase in the frequency of embryos with NTDs, and a decrease in the level of Folbp1 mRNA. CONCLUSIONS: These results demonstrate that alterations in expression of Folbp1 by perturbing either the coding sequence or a critical regulatory cis-element can play a role in NTDs. Published 2003 Wiley-Liss, Inc.
引用
收藏
页码:475 / 487
页数:13
相关论文
共 74 条
[1]  
[Anonymous], 1991, Lancet, V338, P131, DOI 10.1016/0140-6736(91)90133-A
[2]  
ANTONY AC, 1992, BLOOD, V79, P2807
[3]   Folate receptors [J].
Antony, AC .
ANNUAL REVIEW OF NUTRITION, 1996, 16 :501-521
[4]  
Antony AC, 2000, TERATOLOGY, V62, P42, DOI 10.1002/1096-9926(200007)62:1&lt
[5]  
42::AID-TERA9&gt
[6]  
3.0.CO
[7]  
2-U
[8]   Investigation of folate pathway gene polymorphisms and the incidence of neural tube defects in a Texas Hispanic population [J].
Barber, R ;
Shalat, S ;
Hendricks, K ;
Joggerst, B ;
Larsen, R ;
Suarez, L ;
Finnell, R .
MOLECULAR GENETICS AND METABOLISM, 2000, 70 (01) :45-52
[9]  
Barber RC, 1998, AM J MED GENET, V76, P310, DOI 10.1002/(SICI)1096-8628(19980401)76:4<310::AID-AJMG6>3.3.CO
[10]  
2-5