Olfactory receptor neurons exist as distinct subclasses of immature and mature cells in primary culture

A. M. Cunningham; P. B. Manis; R. R. Reed; G. V. Ronnett

doi:10.1016/S0306-4522(99)00193-1

Olfactory receptor neurons exist as distinct subclasses of immature and mature cells in primary culture

A. M. Cunningham, P. B. Manis, R. R. Reed, G. V. Ronnett

School of Medicine

Research output: Contribution to journal › Article › peer-review

29 Scopus citations

Abstract

The processes of neuronal differentiation and survival are key questions in neurobiology. The olfactory system possesses unique regenerative capacity, as its neurons are continually replaced throughout adulthood from a maintained population of precursor cells. Primary cultures of olfactory epithelium enriched in olfactory neurons would provide a useful model to study the processes of neurogenesis, differentiation and senescence. To determine whether immature olfactory neurons could be isolated in primary culture and to investigate the mechanisms underlying these processes, culture conditions which selectively favored the presence of immature olfactory neurons were optimized. Using low plating densities, a population of cells was identified which, by reverse transcription-polymerase chain reaction, demonstrated messages for olfactory neuronal markers, including G(olf), olfactory cyclic nucleotide-gated channel and olfactory marker protein, as well as the p75 low-affinity nerve growth factor receptor. Immunocytochemical analysis showed that these putative immature olfactory neurons possessed immunoreactivity to G(olf), neuron-specific tubulin, neural cell adhesion molecule, synaptophysin and neurofilament. These neurons were defined as olfactory receptor neuron-1 cells. Under these conditions, a separate class of rarely occurring cells with different morphology demonstrated immunoreactivity to mature markers, such as adenylyl cyclase III and olfactory marker protein. Electrophysiologically, these cells displayed properties consistent with those of acutely dissociated olfactory receptor neurons. Another class of rarer cells which represented less than 2% of cells in culture demonstrated immunoreactivity to glial fibrillary acidic protein.These cultures can serve as a model for in vitro analysis of olfactory receptor neuronal development and maintenance, and provide a potential substrate for the development of cell lines. Copyright (C) 1999 IBRO.

Original language	English (US)
Pages (from-to)	1301-1312
Number of pages	12
Journal	Neuroscience
Volume	93
Issue number	4
DOIs	https://doi.org/10.1016/S0306-4522(99)00193-1
State	Published - Aug 1999

Keywords

Differentiation
Electrophysiology
Immunocytochemistry
Neurogenesis
Olfaction
Primary culture

ASJC Scopus subject areas

General Neuroscience

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10.1016/S0306-4522(99)00193-1

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@article{43451653ed49444d8e72452c8cecb30d,

title = "Olfactory receptor neurons exist as distinct subclasses of immature and mature cells in primary culture",

abstract = "The processes of neuronal differentiation and survival are key questions in neurobiology. The olfactory system possesses unique regenerative capacity, as its neurons are continually replaced throughout adulthood from a maintained population of precursor cells. Primary cultures of olfactory epithelium enriched in olfactory neurons would provide a useful model to study the processes of neurogenesis, differentiation and senescence. To determine whether immature olfactory neurons could be isolated in primary culture and to investigate the mechanisms underlying these processes, culture conditions which selectively favored the presence of immature olfactory neurons were optimized. Using low plating densities, a population of cells was identified which, by reverse transcription-polymerase chain reaction, demonstrated messages for olfactory neuronal markers, including G(olf), olfactory cyclic nucleotide-gated channel and olfactory marker protein, as well as the p75 low-affinity nerve growth factor receptor. Immunocytochemical analysis showed that these putative immature olfactory neurons possessed immunoreactivity to G(olf), neuron-specific tubulin, neural cell adhesion molecule, synaptophysin and neurofilament. These neurons were defined as olfactory receptor neuron-1 cells. Under these conditions, a separate class of rarely occurring cells with different morphology demonstrated immunoreactivity to mature markers, such as adenylyl cyclase III and olfactory marker protein. Electrophysiologically, these cells displayed properties consistent with those of acutely dissociated olfactory receptor neurons. Another class of rarer cells which represented less than 2% of cells in culture demonstrated immunoreactivity to glial fibrillary acidic protein.These cultures can serve as a model for in vitro analysis of olfactory receptor neuronal development and maintenance, and provide a potential substrate for the development of cell lines. Copyright (C) 1999 IBRO.",

keywords = "Differentiation, Electrophysiology, Immunocytochemistry, Neurogenesis, Olfaction, Primary culture",

author = "Cunningham, {A. M.} and Manis, {P. B.} and Reed, {R. R.} and Ronnett, {G. V.}",

note = "Funding Information: We thank Helen Cho for technical assistance with neuronal culture. We are grateful to Frank Margolis for the gift of OMP antibody, Alan Sharp for the gift of InsP 3 receptor antibodies and John Hemperly for the gift of anti-NCAM antibody. We thank anonymous reviewers for helpful critiques. This work was supported in part by a grant to A.M.C. from The Medical Foundation, The University of Sydney, NSW, Australia, a grant from The National Institute of Mental Health to R.R.R., a USPHS grant NS-02131 and a Johns Hopkins Clinician Scientist Award to G.V.R., NIDCD grants K04-DC00048 and R01 DC00425 to P.B.M., a grant from International Flavors and Fragrances, and the W. M. Keck Foundation. ",

year = "1999",

month = aug,

doi = "10.1016/S0306-4522(99)00193-1",

language = "English (US)",

volume = "93",

pages = "1301--1312",

journal = "Neuroscience",

issn = "0306-4522",

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T1 - Olfactory receptor neurons exist as distinct subclasses of immature and mature cells in primary culture

AU - Cunningham, A. M.

AU - Manis, P. B.

AU - Reed, R. R.

AU - Ronnett, G. V.

N1 - Funding Information: We thank Helen Cho for technical assistance with neuronal culture. We are grateful to Frank Margolis for the gift of OMP antibody, Alan Sharp for the gift of InsP 3 receptor antibodies and John Hemperly for the gift of anti-NCAM antibody. We thank anonymous reviewers for helpful critiques. This work was supported in part by a grant to A.M.C. from The Medical Foundation, The University of Sydney, NSW, Australia, a grant from The National Institute of Mental Health to R.R.R., a USPHS grant NS-02131 and a Johns Hopkins Clinician Scientist Award to G.V.R., NIDCD grants K04-DC00048 and R01 DC00425 to P.B.M., a grant from International Flavors and Fragrances, and the W. M. Keck Foundation.

PY - 1999/8

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N2 - The processes of neuronal differentiation and survival are key questions in neurobiology. The olfactory system possesses unique regenerative capacity, as its neurons are continually replaced throughout adulthood from a maintained population of precursor cells. Primary cultures of olfactory epithelium enriched in olfactory neurons would provide a useful model to study the processes of neurogenesis, differentiation and senescence. To determine whether immature olfactory neurons could be isolated in primary culture and to investigate the mechanisms underlying these processes, culture conditions which selectively favored the presence of immature olfactory neurons were optimized. Using low plating densities, a population of cells was identified which, by reverse transcription-polymerase chain reaction, demonstrated messages for olfactory neuronal markers, including G(olf), olfactory cyclic nucleotide-gated channel and olfactory marker protein, as well as the p75 low-affinity nerve growth factor receptor. Immunocytochemical analysis showed that these putative immature olfactory neurons possessed immunoreactivity to G(olf), neuron-specific tubulin, neural cell adhesion molecule, synaptophysin and neurofilament. These neurons were defined as olfactory receptor neuron-1 cells. Under these conditions, a separate class of rarely occurring cells with different morphology demonstrated immunoreactivity to mature markers, such as adenylyl cyclase III and olfactory marker protein. Electrophysiologically, these cells displayed properties consistent with those of acutely dissociated olfactory receptor neurons. Another class of rarer cells which represented less than 2% of cells in culture demonstrated immunoreactivity to glial fibrillary acidic protein.These cultures can serve as a model for in vitro analysis of olfactory receptor neuronal development and maintenance, and provide a potential substrate for the development of cell lines. Copyright (C) 1999 IBRO.

AB - The processes of neuronal differentiation and survival are key questions in neurobiology. The olfactory system possesses unique regenerative capacity, as its neurons are continually replaced throughout adulthood from a maintained population of precursor cells. Primary cultures of olfactory epithelium enriched in olfactory neurons would provide a useful model to study the processes of neurogenesis, differentiation and senescence. To determine whether immature olfactory neurons could be isolated in primary culture and to investigate the mechanisms underlying these processes, culture conditions which selectively favored the presence of immature olfactory neurons were optimized. Using low plating densities, a population of cells was identified which, by reverse transcription-polymerase chain reaction, demonstrated messages for olfactory neuronal markers, including G(olf), olfactory cyclic nucleotide-gated channel and olfactory marker protein, as well as the p75 low-affinity nerve growth factor receptor. Immunocytochemical analysis showed that these putative immature olfactory neurons possessed immunoreactivity to G(olf), neuron-specific tubulin, neural cell adhesion molecule, synaptophysin and neurofilament. These neurons were defined as olfactory receptor neuron-1 cells. Under these conditions, a separate class of rarely occurring cells with different morphology demonstrated immunoreactivity to mature markers, such as adenylyl cyclase III and olfactory marker protein. Electrophysiologically, these cells displayed properties consistent with those of acutely dissociated olfactory receptor neurons. Another class of rarer cells which represented less than 2% of cells in culture demonstrated immunoreactivity to glial fibrillary acidic protein.These cultures can serve as a model for in vitro analysis of olfactory receptor neuronal development and maintenance, and provide a potential substrate for the development of cell lines. Copyright (C) 1999 IBRO.

KW - Differentiation

KW - Electrophysiology

KW - Immunocytochemistry

KW - Neurogenesis

KW - Olfaction

KW - Primary culture

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JO - Neuroscience

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ER -

Olfactory receptor neurons exist as distinct subclasses of immature and mature cells in primary culture

Abstract

Keywords

ASJC Scopus subject areas

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