title: Metabolic characterization of all-trans-retinoic acid (ATRA)-induced craniofacial development of murine embryos using in vivo proton magnetic resonance spectroscopy.
abstract: OBJECTIVE
  To characterize the abnormal metabolic profile of all-trans-retinoic acid (ATRA)-induced craniofacial development in mouse embryos using proton magnetic resonance spectroscopy (1H-MRS).
  
  
  METHODS
  Timed-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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  This 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.
authors.affiliation: Cleft Lip and Palate Treatment Center, Second Affiliated Hospital, Shantou University Medical College, Shantou, Guangdong Province, People's Republic of China.
authors.forename: Feifei
authors.initials: F
authors.lastname: Qin
authors.affiliation: Department of Medical Imaging, Second Affiliated Hospital, Shantou University Medical College, Shantou, Guangdong Province, People's Republic of China.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Shen
authors.affiliation: Cleft Lip and Palate Treatment Center, Second Affiliated Hospital, Shantou University Medical College, Shantou, Guangdong Province, People's Republic of China.
authors.forename: Lihong
authors.initials: L
authors.lastname: Peng
authors.affiliation: Department of Medical Imaging, Second Affiliated Hospital, Shantou University Medical College, Shantou, Guangdong Province, People's Republic of China.
authors.forename: Renhua
authors.initials: R
authors.lastname: Wu
authors.affiliation: Department of Plastic and Burn Surgery, Guangzhou Red Cross Hospital, Guangzhou, Guangdong Province, People's Republic of China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Hu
authors.affiliation: Department of Medical Imaging, Second Affiliated Hospital, Shantou University Medical College, Shantou, Guangdong Province, People's Republic of China.
authors.forename: Guishan
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Cleft Lip and Palate Treatment Center, Second Affiliated Hospital, Shantou University Medical College, Shantou, Guangdong Province, People's Republic of China.
authors.forename: Shijie
authors.initials: S
authors.lastname: Tang
mesh_terms: D000014:Abnormalities, Drug-Induced / Q000196:embryology / Q000209:etiology / Q000378:metabolism*; D000818:Animals; D000970:Antineoplastic Agents / Q000633:toxicity; D002794:Choline / Q000378:metabolism; D002972:Cleft Palate / Q000139:chemically induced / Q000196:embryology / Q000378:metabolism; D019465:Craniofacial Abnormalities / Q000139:chemically induced / Q000196:embryology / Q000378:metabolism*; D003401:Creatine / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian / Q000002:abnormalities / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D008055:Lipids / Q000032:analysis; D008297:Male; D051379:Mice; D011247:Pregnancy; D066244:Proton Magnetic Resonance Spectroscopy / Q000379:methods*; D013654:Taurine / Q000378:metabolism; D014212:Tretinoin / Q000633:toxicity*
chemical_list: D000970:Antineoplastic Agents; D008055:Lipids; D013654:Taurine; D014212:Tretinoin; D003401:Creatine; D002794:Choline