Differentiation of mesencephalic progenitor cells into dopaminergic neurons by cytokines.
Rat progenitor cells from the germinal region of the fetal mesencephalon were isolated and expanded in media containing the mitogen epidermal growth factor. These cells remained mitotically active (up to 8 months), were immunoreactive for the progenitor cell marker nestin, and were readily infected with the BAG alpha retrovirus. When incubated in complete media containing serum in poly-L-lysine-coated plates, these cells spontaneously converted to neurons and glia but rarely expressed the dopamine (DA) neuron phenotype. Nineteen different cytokines were screened for their ability to induce the DA phenotype and only interleukin (IL)-1 was found to induce the expression of the DA neuron marker tyrosine hydroxylase (TH). The addition of IL-1, IL-11, leukemia inhibitory factor (LIF), and glial cell line-derived neurotrophic factor (GDNF) were found to further increase the number of TH immunoreactive (TH-ir) cells. The addition of mesencephalic membrane fragments and striatal culture-conditioned media along with the cytokine mixture induced the expression of morphologically mature TH-ir cells that were also immunoreactive for dopa-decarboxylase, the DA transporter, and DA itself. The DA neuron cell counts were approximately 20-25% of the overall cell population and 50% of the neurofilament population. Astrocytes and oligodendrocytes were also present. These data suggest that hematopoietic cytokines participate in the development of the DA neuron phenotype. Parallels between the function of hematopoietic cytokines in bone marrow and the central nervous system may exist and be useful in understanding the factors which regulate the differentiation of neurons in the brain.
Ling
Z D
ZD
Department of Pharmacology, Rush-Presbyterian St. Luke's Medical Center, Chicago, Illinois 60612, USA.
Potter
E D
ED
Lipton
J W
JW
Carvey
P M
PM
D000818:Animals; D001253:Astrocytes / Q000166:cytology; D015415:Biomarkers; D002352:Carrier Proteins / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002462:Cell Membrane / Q000502:physiology; D002478:Cells, Cultured; D003342:Corpus Striatum / Q000502:physiology; D017077:Culture Media, Conditioned; D016207:Cytokines / Q000494:pharmacology*; D004296:Dopa Decarboxylase / Q000096:biosynthesis; D004298:Dopamine / Q000378:metabolism*; D050483:Dopamine Plasma Membrane Transport Proteins; D004622:Embryo, Mammalian; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004921:Erythropoietin / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D051100:Glial Cell Line-Derived Neurotrophic Factor; D006131:Growth Inhibitors / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D017370:Interleukin-11 / Q000494:pharmacology; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines / Q000494:pharmacology; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D008636:Mesencephalon / Q000166:cytology / Q000196:embryology*; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins / Q000494:pharmacology; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D009836:Oligodendroglia / Q000166:cytology; D051381:Rats; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects; D014446:Tyrosine 3-Monooxygenase / Q000096:biosynthesis
D015415:Biomarkers; D002352:Carrier Proteins; D017077:Culture Media, Conditioned; D016207:Cytokines; D050483:Dopamine Plasma Membrane Transport Proteins; C495041:Gdnf protein, rat; D051100:Glial Cell Line-Derived Neurotrophic Factor; D006131:Growth Inhibitors; D007375:Interleukin-1; D017370:Interleukin-11; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D016222:Fibroblast Growth Factor 2; D004921:Erythropoietin; D004815:Epidermal Growth Factor; D014446:Tyrosine 3-Monooxygenase; D004296:Dopa Decarboxylase; D004298:Dopamine