Erythropoietin and bone morphogenetic protein 7 mediate ascorbate-induced dopaminergic differentiation from embryonic mesencephalic precursors.
Mesencephalic precursors derived from early (embryonic day 12; E12) rat embryos were grown in vitro using mitogen basic fibroblast growth factor (bFGF) and these cells efficiently differentiated into dopaminergic (DA) neurons. However, this in vitro DA differentiation was poor in mesencephalic precursors isolated from later embryos (E13-15). Ascorbate (AA) treatment enhanced yields of DA neurons from E12 precursors, and increased the number of DA neurons generated from E13 precursors to levels attained when using E12 precursors. AA markedly up-regulated expression of bone morphogenetic protein 7 (BMP7) and erythropoietin (Epo) in precursors, but did not affect expression of a number of genes known to regulate midbrain DA development. The addition of these recombinant proteins or blockers revealed that both BMP7 and Epo mediate AA-induced DA neuron differentiation.
Lee
Ji-Yeon
JY
Department of Biochemistry, College of Medicine, Hanyang University, Seoul 133-791, Korea.
Koh
Hyun Chul
HC
Chang
Mi-Yoon
MY
Park
Chang-Hwan
CH
Lee
Yong-Sung
YS
Lee
Sang-Hun
SH
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D001205:Ascorbic Acid / Q000494:pharmacology*; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins / Q000502:physiology*; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D004298:Dopamine; D004299:Dopamine beta-Hydroxylase / Q000378:metabolism; D004622:Embryo, Mammalian; D004921:Erythropoietin / Q000502:physiology*; D005260:Female; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D015151:Immunoblotting / Q000379:methods; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D008636:Mesencephalon / Q000187:drug effects / Q000196:embryology / Q000378:metabolism; D011247:Pregnancy; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D016212:Transforming Growth Factor beta; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
D000975:Antioxidants; C525821:Bmp7 protein, rat; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins; D005904:Glial Fibrillary Acidic Protein; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D004921:Erythropoietin; D014446:Tyrosine 3-Monooxygenase; D004299:Dopamine beta-Hydroxylase; D001205:Ascorbic Acid; D004298:Dopamine