Differential gene profiles in developing embryo and fetus after in utero exposure to ethanol.
Alcohol consumption during pregnancy results in morphological abnormalities in the fetuses of humans and experimental animals, and is referred to as fetal alcohol syndrome (FAS). However, the molecular mechanism underlying FAS has not been completely elucidated. The aim of the present study was to investigate the potential molecular mechanisms of ethanol-induced FAS in the developing embryo and fetus. cDNA microarray analysis was used to screen for altered gene profiles. Ethanol at a teratogenic dosage (3.8 g/kg, twice a day) was administered intraperitoneally to pregnant C57Bl/6J mice from gestation day (GD) 6 to 8. Morphologic observations showed excessive malformations of the craniofacial regions (reduction of the face, the absence of eyes, nose, jaw, and mandible, underdevelopment of vibrissae areas, cleft lip, and palate) in ethanol-exposed embryos (GD 10) and fetusus (GD 15). cDNA microarray analysis showed alterations in several gene profiles, including the "palate, lung, and nasal epithelium clone (plunc), "neurofilament, " and "pale ear. " Of these genes, the expressions of plunc were confirmed by reverse-transcription polymerase chain reaction (RT-PCR) and whole-mount in situ hybridization. The plunc was highly expressed in the craniofacial region, specifically in upper airways and nasopharyngeal epithelium. RT-PCR analysis revealed that normal plunc mRNA expression levels were present in GD 15 fetuses, but not in GD 10 embryos. Interestingly, ethanol significantly downregulated the plunc expression in GD 15 fetuses. Our results suggest that ethanol-induced FAS is due in part to the downregulation of plunc expression in the fetus, and this gene may be a candidate biological marker for FAS.
Da Lee
Rhee
R
Department of Toxicology, National Institute of Toxicological Research, Korea Food and Drug Administration, Seoul, Korea.
Rhee
Gyu Seek
GS
An
Sang Mi
SM
Kim
Soon Sun
SS
Kwack
Seung Jun
SJ
Seok
Ji Hyun
JH
Chae
Soo Yeong
SY
Park
Chul Hoon
CH
Yoon
Hyo Jung
HJ
Cho
Dae Hyun
DH
Kim
Hyung Sik
HS
Park
Kui Lea
KL
D000818:Animals; D002492:Central Nervous System Depressants / Q000633:toxicity*; D019465:Craniofacial Abnormalities / Q000139:chemically induced* / Q000662:veterinary; D015536:Down-Regulation; D047108:Embryonic Development / Q000187:drug effects; D000431:Ethanol / Q000633:toxicity*; D005260:Female; D063647:Fetal Alcohol Spectrum Disorders / Q000503:physiopathology*; D047109:Fetal Development / Q000187:drug effects; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D006023:Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D007263:Infusions, Parenteral; D051379:Mice; D008810:Mice, Inbred C57BL; D020411:Oligonucleotide Array Sequence Analysis; D010750:Phosphoproteins / Q000096:biosynthesis* / Q000235:genetics; D011247:Pregnancy; D011297:Prenatal Exposure Delayed Effects; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction
C119074:Bpifa1 protein, mouse; D002492:Central Nervous System Depressants; D006023:Glycoproteins; D010750:Phosphoproteins; D012333:RNA, Messenger; D000431:Ethanol