title: LIF removal increases CRABPI and CRABPII transcripts in embryonic stem cells cultured in retinol or 4-oxoretinol.
abstract: Murine embryonic stem (ES) cells cultured without leukemia inhibitory factor (LIF) or with retinoids differentiate and concomitantly metabolize retinol (vitamin A) to 4-oxoretinol. Our objective was to examine the effects of retinol or 4-oxoretinol on cellular retinoic acid binding protein (CRABP) I and II mRNA levels and retinol metabolism. ES cells were cultured with or without LIF, and with various doses of all-trans-retinol, all-trans-4-oxoretinol, or all-trans-retinoic acid (RA). In ES cells treated with retinol or 4-oxoretinol in the absence of LIF the CRABP-I (Crabp1, NM_013496; GI:7304974) and CRABP-II (Crabp2, NM_007759; GI:33469074) mRNA levels at 72h were 66+/-4 and 413+/-6 fold higher, respectively, than the levels in control ES cells cultured without retinoids and in the presence of LIF. The increase in CRABPI mRNA occurred through an increase in CRABPI gene transcription. CRABPI protein was also increased by >50-fold in cells treated with retinol in the absence of LIF. However [(3)H]4-oxoretinol does not bind to murine CRABPI or CRABPII. CYP26A1 mRNA levels and [(3)H]4-oxoretinol production from [(3)H]retinol increased in cells cultured without LIF and with exogenous retinoids. The enormous increases in CRABPI and II transcripts ( approximately 60 and 400-fold, respectively) in the absence of LIF may regulate aspects of the ES cell differentiation program in response to retinol.
authors.affiliation: Department of Human Ecology, Division of Nutritional Sciences, The University of Texas at Austin, GEA 117, A2700 Austin, TX 78712, USA. mlane@mail.utexas.edu
authors.forename: Michelle A
authors.initials: MA
authors.lastname: Lane
authors.forename: Juliana
authors.initials: J
authors.lastname: Xu
authors.forename: Elana W
authors.initials: EW
authors.lastname: Wilen
authors.forename: Renia
authors.initials: R
authors.lastname: Sylvester
authors.forename: Fadila
authors.initials: F
authors.lastname: Derguini
authors.forename: Lorraine J
authors.initials: LJ
authors.lastname: Gudas
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D053595:Embryonic Stem Cells / Q000187:drug effects* / Q000378:metabolism; D004790:Enzyme Induction / Q000187:drug effects; D053605:Leukemia Inhibitory Factor / Q000378:metabolism*; D051379:Mice; D011485:Protein Binding; D012333:RNA, Messenger / Q000378:metabolism*; D018168:Receptors, Retinoic Acid / Q000235:genetics / Q000378:metabolism*; D000072516:Retinoic Acid 4-Hydroxylase; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects*; D014162:Transfection; D014212:Tretinoin / Q000494:pharmacology; D015854:Up-Regulation; D014801:Vitamin A / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology
chemical_list: D053605:Leukemia Inhibitory Factor; C507838:Lif protein, mouse; D012333:RNA, Messenger; D018168:Receptors, Retinoic Acid; C082927:retinoic acid binding protein I, cellular; C082928:retinoic acid binding protein II, cellular; D014801:Vitamin A; D014212:Tretinoin; C041819:4-oxoretinol; D003577:Cytochrome P-450 Enzyme System; C000607080:Cyp26a1 protein, mouse; D000072516:Retinoic Acid 4-Hydroxylase