Document Detail


Generation of neurons from a nonneuronal precursor in adult olfactory epithelium in vitro.
MedLine Citation:
PMID:  9929609     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Within the olfactory epithelium is a stem cell which can divide and differentiate to produce new sensory neurons. The identity of the neuronal stem cell is unknown but one candidate is the horizontal basal cell which lies adjacent to the basement membrane and expresses keratin. Previous attempts to generate mature sensory neurons from purified horizontal basal cells in vitro were unsuccessful. We show here for the first time that olfactory neurogenesis can be reproduced in vitro from partially-purified cultures of adult rat precursor cells cultivated in a serum-free medium. Rat olfactory epithelium was dissected from the nasal septum and separated from the underlying lamina propria, and its cells were dissociated and grown in a medium containing epidermal growth factor for 5 days. Immunochemistry showed that only supporting cells (SUS1-positive) and horizontal basal cells (keratin-positive) survived for this period. At day 6, the cells were stressed either by passaging them or by a simple mechanical stress. In each case, a morphological and immunological differentiation was observed within 24-48 hr. Newly formed bipolar cells were found to be S100-, glial fibrillary acidic protein (GFAP-), neural cell adhesion molecule (N-CAM+), and/or microtubule-associated protein 5 (MAP-5+). After passaging 14% of the surviving cells were immature neurons (MAP-5+) and 4% were mature olfactory neurons (MAP-5+) and olfactory marker protein (OMP+)). In addition the same experiment was conducted on transgenic mice in which the lacZ gene was linked to the OMP promoter. Using 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-Gal) staining we showed that OMP+ cells disappeared before day 5 in culture but reappeared after passaging. These results suggest that olfactory sensory neurons can arise from a non-neuronal precursor, probably the keratin-positive horizontal basal cell.
Authors:
G Sicard; F Féron; J L Andrieu; A Holley; A Mackay-Sim
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Annals of the New York Academy of Sciences     Volume:  855     ISSN:  0077-8923     ISO Abbreviation:  Ann. N. Y. Acad. Sci.     Publication Date:  1998 Nov 
Date Detail:
Created Date:  1999-02-24     Completed Date:  1999-02-24     Revised Date:  2003-11-14    
Medline Journal Info:
Nlm Unique ID:  7506858     Medline TA:  Ann N Y Acad Sci     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  223-5     Citation Subset:  IM    
Affiliation:
Université Claude Bernard, Villeurbanne, France.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Differentiation
Cells, Cultured
Epithelial Cells / cytology*
Mice
Nerve Tissue Proteins / biosynthesis
Olfactory Mucosa / cytology
Olfactory Receptor Neurons / cytology*,  metabolism
Rats
Stem Cells / cytology
Chemical
Reg. No./Substance:
0/Nerve Tissue Proteins

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