Human olfactory epithelial cells generated in vitro express diverse neuronal characteristics

• Published in: Neuroscience Vol. 158; no. 2; pp. 642 - 653
• Main Authors: Borgmann-Winter, K.E, Rawson, N.E, Wang, H.-Y, Wang, H, MacDonald, M.L, Ozdener, M.H, Yee, K.K, Gomez, G, Xu, J, Bryant, B, Adamek, G, Mirza, N, Pribitkin, E, Hahn, C.-G
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Ltd 23.01.2009; Elsevier
• Subjects: 5-HT receptor; Adult; Animals; Biological and medical sciences; Cells, Cultured; Dopamine Agents - pharmacology; dopamine receptor; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Female; Fundamental and applied biological sciences. Psychology; Gene Expression - physiology; Glycine - pharmacology; Humans; Immunoprecipitation - methods; In Vitro Techniques; Male; Middle Aged; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; neurogenesis; Neurology; neuronal cell culture; NMDA receptor; odorant receptor; Olfactory Marker Protein - genetics; Olfactory Marker Protein - metabolism; Olfactory Mucosa - cytology; Olfactory Receptor Neurons - metabolism; Rats; Rats, Sprague-Dawley; Receptors, N-Methyl-D-Aspartate - genetics; Receptors, N-Methyl-D-Aspartate - metabolism; Receptors, Odorant - genetics; Receptors, Odorant - metabolism; Receptors, Serotonin - genetics; Receptors, Serotonin - metabolism; RNA, Messenger - metabolism; Serotonin Agents - pharmacology; Vertebrates: nervous system and sense organs; Young Adult; GTP; digoxigenin; adenylate cyclase type 3; olfactory marker protein; phosphate-buffered saline; dopamine receptor; N-methyl-D-aspartic acid receptor; OcNc1; serotonin; N-methyl-D-aspartic acid; RT-PCR; neurogenesis; NMDA; 5-HT receptor; PBS; DIG; phosphate-buffered saline containing 0.3% Tween-20; D2R; reverse transcriptase PCR; human olfactory epithelial; ethylene diamine tetraacetic acid; neuronal cell culture; guanosine triphosphate; olfactory sensory neuron; OSN; EDTA; NR1; NMDAR1; PLCγ1; EGTA; OMP; ACIII; PBST; olfactory cyclic nucleotide gated channel 1; NMDAR; phospholipase C-γ1; NMDA receptor; 5-HT; ethylene glycol tetraacetic acid; odorant receptor; NMDAR2A/2B; NR2A/2B; hOE; Human; Dopamine receptor; Epithelial cell; Glutamate receptor; Serotonine receptor; In vitro; Neurogenesis; Biological receptor
• ISSN: 0306-4522; 1873-7544
• DOI: 10.1016/j.neuroscience.2008.09.059

Abstract The olfactory epithelium constitutes the sole source of regenerating neural cells that can be obtained from a living human. As such, primary cultures derived from human olfactory epithelial biopsies can be utilized to study neurobiological characteristics of individuals under different conditions and disease states. Here, using such human cultures, we report in vitro generation of cells that exhibit a complex neuronal phenotype, encompassing receptors and signaling pathways pertinent to both olfaction and other aspects of CNS function. Using in situ hybridization, we demonstrate for the first time the native expression of olfactory receptors in cultured cells derived from human olfactory epithelial tissue. We further establish the presence and function of olfactory transduction molecules in these cells using immunocytochemistry, calcium imaging and molecular methods. Western blot analysis revealed the expression of neurotransmitter receptors for dopamine (D2R), 5-HT (5HT2C) and NMDA subtypes 1 and 2A/2B. Stimulation with dopamine or 5-HT enhanced receptor G protein activation in a subtype specific manner, based on 35S-guanosine triphosphate incorporation assay. Functional characteristics of the cultured cells are demonstrated through enhanced tyrosine phosphorylation of NMDAR 2A/2B and recruitment of signaling partners in response to NMDA stimulation. The array of neuronal characteristics observed here establishes that proliferating cells derived from the human olfactory epithelium differentiate in vitro to express functional and molecular attributes of mature olfactory neurons. These cultured neural cells exhibit neurotransmitter pathways important in a number of neuropsychiatric disorders. Their ready availability from living humans thus provides a new tool to link functional and molecular features of neural cells with clinical characteristics of individual living patients.