Human olfactory neuroepithelial cells: tyrosine phosphorylation and process extension are increased by the combination of IL-1beta, IL-6, NGF, and bFGF.
Olfactory neuroepithelial cells (ONC) grown from biopsies of human donors are a novel cell culture system that may facilitate studies into normal and disease-related human neurobiology. We further characterized the expression of cell surface markers and intermediate filaments, and responses to neurotrophic factors by ONC. ONC are positive for cell surface markers N-CAM, PSA-N-CAM, neutral endopeptidase, N-aminopeptidase, NGF low-affinity receptor homologue (CD40), and transferrin receptor by flow cytometry for the intermediate filament proteins peripherin, vimentin, and NF-H by immunocytochemistry. Responses to neurotrophic factors measured were process outgrowth, cytoskeletal protein expression, and protein phosphorylation. Process outgrowth was increased by interleukin-beta 164-171 (IL-1beta) or by the combination of IL-1beta, interleukin-6 (IL-6), nerve growth factor (NGF), and basic fibroblast growth factor (bFGF). This combination of IL-1beta, IL-6, NGF, and bFGF (16NF) increased expression of two cytoskeletal proteins, NF-H protein and microtubule-associated protein tau. Application of the individual neurotrophic factors IL-1beta, IL-6, NGF, and bFGF increased protein phosphorylation, while 16NF produced an immediate increase in tyrosine phosphorylation of several proteins (MW of 40-80, 120, 150, and 190 kDa). The 16NF combination appears to act through a tyrosine-kinase-mediated pathway to induce process extension and increase NF-H expression. The ONC culture has the potential to be further explored to examine the relationship among process outgrowth, protein phosphorylation, and synergy between neurotrophin and cytokine receptor systems.
Vawter
M P
MP
Section on Preclinical Neuroscience, NIMH, Washington, DC 20032, USA.
Basaric-Keys
J
J
Li
Y
Y
Lester
D S
DS
Lebovics
R S
RS
Lesch
K P
KP
Kulaga
H
H
Freed
W J
WJ
Sunderland
T
T
Wolozin
B
B
D015703:Antigens, CD / Q000032:analysis; D000944:Antigens, Differentiation, B-Lymphocyte / Q000032:analysis; D015415:Biomarkers; D018826:CD13 Antigens / Q000032:analysis; D019013:CD40 Antigens / Q000032:analysis; D002454:Cell Differentiation / Q000187:drug effects; D003598:Cytoskeletal Proteins / Q000187:drug effects / Q000378:metabolism; D004848:Epithelium / Q000737:chemistry / Q000187:drug effects / Q000201:enzymology; D005136:Eye Proteins / Q000032:analysis; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D005434:Flow Cytometry; D019922:GAP-43 Protein; D019204:GTP-Binding Proteins / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000494:pharmacology*; D015850:Interleukin-6 / Q000494:pharmacology*; D007381:Intermediate Filament Proteins / Q000032:analysis; D008562:Membrane Glycoproteins / Q000032:analysis; D008869:Microtubule-Associated Proteins / Q000032:analysis; D015260:Neprilysin / Q000032:analysis; D009414:Nerve Growth Factors / Q000494:pharmacology*; D009419:Nerve Tissue Proteins / Q000032:analysis; D019006:Neural Cell Adhesion Molecules / Q000032:analysis; D016900:Neurofilament Proteins / Q000032:analysis; D009479:Neuropeptides / Q000032:analysis; D009831:Olfactory Mucosa; D009832:Olfactory Nerve; D009833:Olfactory Pathways / Q000166:cytology* / Q000201:enzymology; D018034:Olfactory Receptor Neurons; D064531:Peripherins; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011990:Receptors, Transferrin; D015398:Signal Transduction / Q000502:physiology; D014443:Tyrosine / Q000187:drug effects / Q000378:metabolism*; D014746:Vimentin / Q000032:analysis
D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; D015415:Biomarkers; D019013:CD40 Antigens; C068126:CD71 antigen; D003598:Cytoskeletal Proteins; D005136:Eye Proteins; D019922:GAP-43 Protein; D007375:Interleukin-1; D015850:Interleukin-6; D007381:Intermediate Filament Proteins; D008562:Membrane Glycoproteins; D008869:Microtubule-Associated Proteins; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D019006:Neural Cell Adhesion Molecules; D016900:Neurofilament Proteins; D009479:Neuropeptides; C578994:PRPH protein, human; D064531:Peripherins; D011990:Receptors, Transferrin; D014746:Vimentin; C044882:neurofilament protein L; D016222:Fibroblast Growth Factor 2; C051774:neurofilament protein H; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases; D018826:CD13 Antigens; D015260:Neprilysin; D019204:GTP-Binding Proteins