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.
Section on Preclinical Neuroscience, NIMH, Washington, DC 20032, USA.
M P
MP
Vawter
J
J
Basaric-Keys
Y
Y
Li
D S
DS
Lester
R S
RS
Lebovics
K P
KP
Lesch
H
H
Kulaga
W J
WJ
Freed
T
T
Sunderland
B
B
Wolozin
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