Influence of LIF and BMP-2 on differentiation and development of glial cells in primary cultures of embryonic rat cerebral hemisphere.
Cells prepared from the cerebral hemisphere of embryonic Day 18 rats were maintained for 2 days in serum-free modified Bottenstein-Sato (mBS) medium containing thyroid hormone (TH), with or without leukemia inhibitory factor (LIF) or bone morphogenetic protein (BMP)-2, and these influences on the differentiation and development of glial cells were investigated using the cells maintained in mBS medium containing TH as controls. The levels of glial fibrillary acidic protein (GFAP) expression and the number of GFAP-positive astrocytes increased markedly with the addition of LIF or BMP-2, and were enhanced further with the addition of both LIF and BMP-2. The number of O1-positive oligodendrocytes increased with the addition of LIF, whereas it decreased with the addition of BMP-2. The number did not change with the addition of both cytokines. Using antibody against platelet-derived growth factor (PDGF), we then excluded indirect effects of these cytokines through PDGF, which would increase by accelerated astrocyte development. When PDGF was neutralized, the number of oligodendrocytes increased under all conditions examined. As a result of the neutralization, the effect of BMP-2 on oligodendrocyte differentiation was eliminated, although LIF remained effective. These results suggest that the differentiation of oligodendrocytes was delayed partially by PDGF even in control cultures. It is also suggested that LIF and BMP-2, each of which accelerates the differentiation and development of astrocytes, would seem to have different effects on oligodendrocyte differentiation, i.e., LIF would directly affect oligodendrocyte differentiation, whereas BMP-2 would affect it mainly through PDGF.
Adachi
Tatsumi
T
Department of Basic Medical Sciences, National Institute for Minamata Disease, Minamata, Kumamoto, Japan. taadachi@nimd.go.jp
Takanaga
Hiromi
H
Kunimoto
Manabu
M
Asou
Hiroaki
H
D000704:Analysis of Variance; D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D015153:Blotting, Western / Q000379:methods; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology*; D002452:Cell Count / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology*; D017077:Culture Media, Conditioned / Q000494:pharmacology; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004347:Drug Interactions; D004622:Embryo, Mammalian; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D007211:Indoles / Q000378:metabolism; D015850:Interleukin-6 / Q000494:pharmacology*; D053605:Leukemia Inhibitory Factor; D008297:Male; D009457:Neuroglia / Q000166:cytology / Q000187:drug effects*; D010982:Platelet-Derived Growth Factor / Q000276:immunology / Q000378:metabolism; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
D000906:Antibodies; C525720:Bmp2 protein, rat; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free; D005904:Glial Fibrillary Acidic Protein; D007211:Indoles; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D010982:Platelet-Derived Growth Factor; D016212:Transforming Growth Factor beta; C007293:DAPI; D014446:Tyrosine 3-Monooxygenase