Fetal alcohol syndrome (FAS) in C57BL/6 mice detected through proteomics screening of the amniotic fluid.
BACKGROUND
Fetal Alcohol Syndrome (FAS), a severe consequence of the Fetal Alcohol Spectrum Disorders, is associated with craniofacial defects, mental retardation, and stunted growth. Previous studies in C57BL/6J and C57BL/6N mice provide evidence that alcohol-induced pathogenesis follows early changes in gene expression within specific molecular pathways in the embryonic headfold. Whereas the former (B6J) pregnancies carry a high-risk for dysmorphogenesis following maternal exposure to 2.9 g/kg alcohol (two injections spaced 4.0 h apart on gestation day 8), the latter (B6N) pregnancies carry a low-risk for malformations. The present study used this murine model to screen amniotic fluid for biomarkers that could potentially discriminate between FAS-positive and FAS-negative pregnancies.


METHODS
B6J and B6N litters were treated with alcohol (exposed) or saline (control) on day 8 of gestation. Amniotic fluid aspirated on day 17 (n = 6 replicate litters per group) was subjected to trypsin digestion for analysis by matrix-assisted laser desorption-time of flight mass spectrometry with the aid of denoising algorithms, statistical testing, and classification methods.


RESULTS
We identified several peaks in the proteomics screen that were reduced consistently and specifically in exposed B6J litters. Preliminary characterization by liquid chromatography tandem mass spectrometry and multidimensional protein identification mapped the reduced peaks to alpha fetoprotein (AFP). The predictive strength of AFP deficiency as a biomarker for FAS-positive litters was confirmed by area under the receiver operating characteristic curve.


CONCLUSIONS
: These findings in genetically susceptible mice support clinical observations in maternal serum that implicate a decrease in AFP levels following prenatal alcohol damage.
Datta
Susmita
S
Department of Bioinformatics and Biostatistics, School of Public Health and Information Science, University of Louisville, Louisville, Kentucky 40292, USA. susmita.datta@louisville.edu
Turner
Delano
D
Singh
Reetu
R
Ruest
L Bruno
LB
Pierce
William M
WM
Knudsen
Thomas B
TB
D000653:Amniotic Fluid / Q000737:chemistry*; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D000013:Congenital Abnormalities / Q000209:etiology; D000431:Ethanol; D005260:Female; D063647:Fetal Alcohol Spectrum Disorders / Q000175:diagnosis* / Q000473:pathology; D005327:Fetal Resorption / Q000209:etiology; D020567:Fetal Weight / Q000187:drug effects; D008098:Litter Size; D018811:Maternal Exposure / Q000009:adverse effects; D051379:Mice; D008810:Mice, Inbred C57BL; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D040901:Proteomics; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D000509:alpha-Fetoproteins / Q000032:analysis*
D015415:Biomarkers; D000509:alpha-Fetoproteins; D000431:Ethanol