QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

HOME  |   SEARCH  |   ARCHIVE  |   SUBSCRIBE  |   CONTACT  |   HELP

This Article Full Text Full Text (PDF) Submit an eLetter Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (19) Citing Articles via Google Scholar Google Scholar Articles by Barber, R. D. Articles by Ronnett, G. V. Search for Related Content PubMed PubMed Citation Articles by Barber, R. D. Articles by Ronnett, G. V.

 Previous Article  |  Next Article 

The Journal of Neuroscience, May 15, 2000, 20(10):3695-3704

Isolation and In Vitro Differentiation of Conditionally Immortalized Murine Olfactory Receptor Neurons

Robert D. Barber^{1, 2}, Donna E. Jaworsky², King-Wai Yau^{1, 2}, and Gabriele V. Ronnett^{1, 3}

¹ Howard Hughes Medical Institute and Departments of ² Neuroscience and ³ Neurology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205

Two major challenges exist in our understanding of the olfactory system. One concerns the enormous combinatorial code underlying odorant discrimination by odorant receptors. The other relates to neurogenesis and neuronal development in the olfactory epithelium. To address these issues, continuous cell cultures containing olfactory receptor neurons (ORNs) were obtained from olfactory epithelia of H-2K^b-tsA58 transgenic mice. ORNs were detected and characterized by immunocytochemistry, RT-PCR, and Western blot for the markers G_olf, adenylyl cyclase III, the olfactory cyclic nucleotide-gated channel subunits, and olfactory marker protein. In culture, epidermal growth factor and nerve growth factor stimulated proliferation, and brain-derived neurotrophic factor and neurotrophin-3 induced cellular maturation.

Clonal cell lines were isolated by fluorescence-activated cell sorting with anti-neural cell adhesion molecule antibodies, and of 144 single cells plated, 39 clones were expanded, propagated, and stored in liquid nitrogen. All attempts at recovery of clonal lines from frozen stocks have been successful. The most thoroughly characterized clone, 3NA12, expressed ORN markers and responded to stimulation by single odorants. Each odorant activated ~1% of cells in a clonal line, and this suggests that many different odorant receptors may be expressed by these clonal cells. Therefore, these cell lines and the method by which they have been obtained represent a significant advance in the generation of olfactory cell cultures and provide a system to investigate odorant coding and olfactory neurogenesis.

Key words: olfactory receptor neurons; H-2K^b-tsA58 transgenic mouse; cell lines; immortalization; proliferation; trophic factors

---

Copyright © 2000 Society for Neuroscience  0270-6474/00/20103695-10$05.00/0

This article has been cited by other articles:

				T. Murata, M. Tsuboi, K. Hikita, and N. Kaneda Protective Effects of Neurotrophic Factors on Tumor Necrosis Factor-related Apoptosis-inducing Ligand (TRAIL)-mediated Apoptosis of Murine Adrenal Chromaffin Cell Line tsAM5D J. Biol. Chem., August 11, 2006; 281(32): 22503 - 22516. [Abstract] [Full Text] [PDF]

				E. Shirokova, K. Schmiedeberg, P. Bedner, H. Niessen, K. Willecke, J.-D. Raguse, W. Meyerhof, and D. Krautwurst Identification of Specific Ligands for Orphan Olfactory Receptors: G PROTEIN-DEPENDENT AGONISM AND ANTAGONISM OF ODORANTS J. Biol. Chem., March 25, 2005; 280(12): 11807 - 11815. [Abstract] [Full Text] [PDF]

				D. Dory, H. Echchannaoui, M. Letiembre, F. Ferracin, J. Pieters, Y. Adachi, S. Akashi, W. Zimmerli, and R. Landmann Generation and functional characterization of a clonal murine periportal Kupffer cell line from H-2Kb -tsA58 mice J. Leukoc. Biol., July 1, 2003; 74(1): 49 - 59. [Abstract] [Full Text] [PDF]

				P. J. Simpson, I. Miller, C. Moon, A. L. Hanlon, D. J. Liebl, and G. V. Ronnett Atrial Natriuretic Peptide Type C Induces a Cell-Cycle Switch from Proliferation to Differentiation in Brain-Derived Neurotrophic Factor- or Nerve Growth Factor-Primed Olfactory Receptor Neurons J. Neurosci., July 1, 2002; 22(13): 5536 - 5551. [Abstract] [Full Text] [PDF]

Home  |   Search  |   Archive  |   Subscribe  |   Contact  |   Help