Metabolic Characterization of All-Trans-Retinoic Acid (ATRA)–Induced Craniofacial Development of Murine Embryos Using In Vivo Proton Magnetic Resonance Spectroscopy

Abstract

AimTo characterize the abnormal metabolic profile of all-trans-retinoic acid (ATRA)–induced craniofacial development in mouse embryos using proton magnetic resonance spectroscopy (1H-MRS).MethodsTimed-pregnant mice were treated by oral gavage on the morning of embryonic gestation day 11 (E11) with all-trans-retinoic acid (ATRA). Dosing solutions were adjusted by maternal body weight to provide 30, 70, or 100 mg/kg RA. The control group was given an equivalent volume of the carrier alone. Using an Agilent 7.0 T MR system and a combination of surface coil coils, a 3 mm×3 mm×3 mm 1H-MRS voxel was selected along the embryonic craniofacial tissue. 1H-MRS was performed with a single-voxel method using PRESS sequence and analyzed using LCModel software. Hematoxylin and eosin was used to detect and confirm cleft palate.Result 1H-MRS revealed elevated choline levels in embryonic craniofacial tissue in the RA70 and RA100 groups compared to controls (P<0.05). Increased choline levels were also found in the RA70 and RA100 groups compared with the RA30 group (P<0.01). High intra-myocellular lipids at 1.30 ppm (IMCL13) in the RA100 group compared to the RA30 group were found (P<0.01). There were no significant changes in taurine, intra-myocellular lipids at 2.10 ppm (IMCL21), and extra-myocellular lipids at 2.30 ppm (EMCL23). Cleft palate formation was observed in all fetuses carried by mice administered 70 and 100 mg/kg RA.ConclusionsThis novel study suggests that the elevated choline and lipid levels found by 1H-MRS may represent early biomarkers of craniofacial defects. Further studies will determine performance of this test and pathogenetic mechanisms of craniofacial malformation.

Prenatal retinoic acid exposure reveals candidate genes for craniofacial disorders

“…Moreover, we revealed here a link between the major OAVS environmental factors (i.e. ATRA exposure 18 , 24 , hypoxia 15 , 38 and diabetes 11 , 12 ) and craniofacial defects.…”

Position-dependent correlation between TBX22 exon 5 methylation and palatal shelf fusion in the development of cleft palate

“…For this study, an ATRA (Sigma-Aldrich, St. Louis., MO, USA)induced mouse cleft palate model was used, and embryonic palatal shelf tissues were resected at E13.5, E14.5, and E16.5, as reported previously (Qin et al 2014, Shu et al 2018. This animal study protocol was approved by the Laboratory Animal Ethical Committee of Medical College of Shantou University (SUMC2015-106; Shantou, China), and experiments were carried out in accordance with the animal care guidelines of the US National Institutes of Health.…”

Correlation between HDAC4 enhancer DNA methylation and mRNA expression during palatal fusion induced by all‐trans retinoic acid

“…The case groups were administered ATRA (Sigma-Aldrich, St. Louis, MO), dissolved in corn oil by gavage at 70 mg/kg, 18 whereas the control group mice were given an equivalent volume of corn oil according to their weight. C57BL/6J mice were purchased from the Beijing Vital River Laboratory Animal Technology Co Ltd (Beijing, China).…”

“…Pregnant females at E10.5 were randomly divided into two groups. The case groups were administered ATRA (Sigma-Aldrich, St. Louis, MO), dissolved in corn oil by gavage at 70 mg/kg, 18 whereas the control group mice were given an equivalent volume of corn oil according to their weight. At E14.5, the mice were killed, and palatal shelves were dissected using microbiological instruments.…”