title: Human olfactory epithelial cells generated in vitro express diverse neuronal characteristics.
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.
authors.affiliation: Department of Psychiatry, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: K E
authors.initials: KE
authors.lastname: Borgmann-Winter
authors.forename: N E
authors.initials: NE
authors.lastname: Rawson
authors.forename: H-Y
authors.initials: HY
authors.lastname: Wang
authors.forename: H
authors.initials: H
authors.lastname: Wang
authors.forename: M L
authors.initials: ML
authors.lastname: Macdonald
authors.forename: M H
authors.initials: MH
authors.lastname: Ozdener
authors.forename: K K
authors.initials: KK
authors.lastname: Yee
authors.forename: G
authors.initials: G
authors.lastname: Gomez
authors.forename: J
authors.initials: J
authors.lastname: Xu
authors.forename: B
authors.initials: B
authors.lastname: Bryant
authors.forename: G
authors.initials: G
authors.lastname: Adamek
authors.forename: N
authors.initials: N
authors.lastname: Mirza
authors.forename: E
authors.initials: E
authors.lastname: Pribitkin
authors.forename: C-G
authors.initials: CG
authors.lastname: Hahn
mesh_terms: D000328:Adult; D000818:Animals; D002478:Cells, Cultured; D015259:Dopamine Agents / Q000494:pharmacology; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D015870:Gene Expression / Q000502:physiology*; D005998:Glycine / Q000494:pharmacology; D006801:Humans; D047468:Immunoprecipitation / Q000379:methods; D066298:In Vitro Techniques; D008297:Male; D008875:Middle Aged; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D051842:Olfactory Marker Protein / Q000235:genetics / Q000378:metabolism; D009831:Olfactory Mucosa / Q000166:cytology*; D018034:Olfactory Receptor Neurons / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D016194:Receptors, N-Methyl-D-Aspartate / Q000235:genetics / Q000378:metabolism; D018035:Receptors, Odorant / Q000235:genetics / Q000378:metabolism; D011985:Receptors, Serotonin / Q000235:genetics / Q000378:metabolism; D018490:Serotonin Agents / Q000494:pharmacology; D055815:Young Adult
chemical_list: D015259:Dopamine Agents; D009419:Nerve Tissue Proteins; C497597:OMP protein, human; D051842:Olfactory Marker Protein; D012333:RNA, Messenger; D016194:Receptors, N-Methyl-D-Aspartate; D018035:Receptors, Odorant; D011985:Receptors, Serotonin; D018490:Serotonin Agents; D005998:Glycine