Induction of cholinergic function in cultured sympathetic neurons by periosteal cells: cellular mechanisms.
Periosteum, the connective tissue surrounding bone, alters the transmitter properties of its sympathetic innervation during development in vivo and after transplantation. Initial noradrenergic properties are downregulated and the innervation acquires cholinergic and peptidergic properties. To elucidate the cellular mechanisms responsible, sympathetic neurons were cultured with primary periosteal cells or osteoblast cell lines. Both primary cells and an immature osteoblast cell line, MC3T3-E1, induced choline acetyltransferase (ChAT) activity. In contrast, lines representing marrow stromal cells or mature osteoblasts did not increase ChAT. Growth of periosteal cells with sympathetic neurons in transwell cultures that prevent direct contact between the neurons and periosteal cells or addition of periosteal cell-conditioned medium to neuron cultures induced ChAT, indicating that periosteal cells release a soluble cholinergic inducing factor. Antibodies against LIFRbeta, a receptor subunit shared by neuropoietic cytokines, prevented ChAT induction in periosteal cell/neuron cocultures, suggesting that a member of this family is responsible. ChAT activity was increased in neurons grown with periosteal cells or conditioned medium from mice lacking either leukemia inhibitory factor (LIF) or LIF and ciliary neurotrophic factor (CNTF). These results provide evidence that periosteal cells influence sympathetic neuron phenotype by releasing a soluble cholinergic factor that is neither LIF nor CNTF but signals via LIFRbeta.
Asmus
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Department of Biochemistry and Molecular Biology, Centre College, Danville, Kentucky 40422, USA.
Tian
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Landis
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D016475:3T3 Cells; D000818:Animals; D000831:Animals, Newborn; D002448:Cell Adhesion; D002478:Cells, Cultured; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D020934:Ciliary Neurotrophic Factor / Q000235:genetics / Q000502:physiology; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D006131:Growth Inhibitors / Q000235:genetics / Q000502:physiology; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; D008222:Lymphokines / Q000235:genetics / Q000502:physiology; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D010006:Osteoblasts / Q000166:cytology; D051381:Rats; D011950:Receptors, Cholinergic / Q000378:metabolism*; D018121:Receptors, Cytokine / Q000378:metabolism; D053606:Receptors, OSM-LIF; D015398:Signal Transduction; D013564:Sympathetic Nervous System / Q000201:enzymology
D020934:Ciliary Neurotrophic Factor; D017077:Culture Media, Conditioned; D006131:Growth Inhibitors; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; C507838:Lif protein, mouse; C507841:Lifr protein, mouse; C507842:Lifr protein, rat; D008222:Lymphokines; D011950:Receptors, Cholinergic; D018121:Receptors, Cytokine; D053606:Receptors, OSM-LIF; D002795:Choline O-Acetyltransferase