title: Expressing exogenous functional odorant receptors in cultured olfactory sensory neurons.
abstract: BACKGROUND
  Olfactory discrimination depends on the large numbers of odorant receptor genes and differential ligand-receptor signaling among neurons expressing different receptors. In this study, we describe an in vitro system that enables the expression of exogenous odorant receptors in cultured olfactory sensory neurons. Olfactory sensory neurons in the culture express characteristic signaling molecules and, therefore, provide a system to study receptor function within its intrinsic cellular environment.
  
  
  RESULTS
  We demonstrate that cultured olfactory sensory neurons express endogenous odorant receptors. Lentiviral vector-mediated gene transfer enables successful ectopic expression of odorant receptors. We show that the ectopically expressed mouse I7 is functional in the cultured olfactory sensory neurons. When two different odorant receptors are ectopically expressed simultaneously, both receptor proteins co-localized in the same olfactory sensory neurons up to 10 days in vitro.
  
  
  CONCLUSIONS
  This culture technique provided an efficient method to culture olfactory sensory neurons whose morphology, molecular characteristics and maturation progression resembled those observed in vivo. Using this system, regulation of odorant receptor expression and its ligand specificity can be studied in its intrinsic cellular environment.
authors.affiliation: Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California 95616, USA. hycchen@ucdavis.edu
authors.forename: Huaiyang
authors.initials: H
authors.lastname: Chen
authors.forename: Sepehr
authors.initials: S
authors.lastname: Dadsetan
authors.forename: Alla F
authors.initials: AF
authors.lastname: Fomina
authors.forename: Qizhi
authors.initials: Q
authors.lastname: Gong
mesh_terms: D000818:Animals; D001369:Axons / Q000502:physiology; D015415:Biomarkers; D002478:Cells, Cultured; D005260:Female; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D018034:Olfactory Receptor Neurons / Q000166:cytology* / Q000502:physiology* / Q000648:ultrastructure; D011247:Pregnancy; D018035:Receptors, Odorant / Q000235:genetics*
chemical_list: D015415:Biomarkers; D018035:Receptors, Odorant
references.citation: J Neurosci. 2007 Jun 20;27(25):6675-83
references.pmid: 17581954
references.citation: Cell Tissue Res. 1999 Dec;298(3):405-14
references.pmid: 10639731
references.citation: J Neurosci. 1999 Nov 15;19(22):9856-64
references.pmid: 10559395
references.citation: J Comp Neurol. 1994 Jun 15;344(3):336-48
references.pmid: 8063958
references.citation: Neuron. 1995 Oct;15(4):779-89
references.pmid: 7576628
references.citation: Science. 1998 Jan 9;279(5348):237-42
references.pmid: 9422698
references.citation: J Neurosci. 2004 Jun 23;24(25):5670-83
references.pmid: 15215289
references.citation: J Neurosci. 2001 Dec 15;21(24):9713-23
references.pmid: 11739580
references.citation: Cell. 2004 Jun 11;117(6):801-15
references.pmid: 15186780
references.citation: Chem Senses. 2004 Sep;29(7):595-616
references.pmid: 15337685
references.citation: Curr Protoc Neurosci. 2001 May;Chapter 3:Unit 3.5
references.pmid: 18428469
references.citation: J Neurosci. 1997 May 1;17(9):3120-7
references.pmid: 9096146
references.citation: Proc Natl Acad Sci U S A. 2002 Jun 25;99(13):9015-20
references.pmid: 12084939
references.citation: J Neurosci. 1991 May;11(5):1243-55
references.pmid: 1851216
references.citation: Neuroscience. 1999;94(1):131-40
references.pmid: 10613503
references.citation: Neuron. 1992 Mar;8(3):441-53
references.pmid: 1550671
references.citation: Proc Natl Acad Sci U S A. 2004 Jan 27;101(4):1069-74
references.pmid: 14732684
references.citation: Cell. 2007 Nov 30;131(5):1009-17
references.pmid: 18045541
references.citation: Brain Res. 1993 Jun 4;613(1):168-72
references.pmid: 8348301
references.citation: Science. 2002 Feb 1;295(5556):868-72
references.pmid: 11786607
references.citation: Nat Neurosci. 2000 Jul;3(7):687-93
references.pmid: 10862701
references.citation: Science. 2003 Dec 19;302(5653):2088-94
references.pmid: 14593185
references.citation: Neuron. 1989 Jul;3(1):115-27
references.pmid: 2482777
references.citation: Brain Res Mol Brain Res. 1997 Sep;48(2):270-8
references.pmid: 9332724
references.citation: Cell. 1998 Dec 23;95(7):917-26
references.pmid: 9875846
references.citation: J Biol Chem. 2007 May 18;282(20):15284-93
references.pmid: 17387175
references.citation: Cell. 1994 Dec 30;79(7):1245-55
references.pmid: 7528109
references.citation: Cell. 2006 Dec 1;127(5):1057-69
references.pmid: 17129788
references.citation: J Neurocytol. 2005 Mar;34(1-2):37-47
references.pmid: 16374708
references.citation: Neuron. 2002 Aug 15;35(4):681-96
references.pmid: 12194868
references.citation: Cell. 1994 Dec 16;79(6):981-91
references.pmid: 8001145
references.citation: Exp Cell Res. 2005 Jun 10;306(2):309-16
references.pmid: 15925585
references.citation: J Neurochem. 2004 Sep;90(6):1453-63
references.pmid: 15341529
references.citation: Dev Neurobiol. 2008 Mar;68(4):476-86
references.pmid: 18214836
references.citation: Science. 1990 Dec 7;250(4986):1403-6
references.pmid: 2255909
references.citation: Chem Senses. 1997 Aug;22(4):467-76
references.pmid: 9279469
references.citation: J Neurocytol. 1997 Oct;26(10):691-706
references.pmid: 9368882
references.citation: Cell. 1999 Mar 5;96(5):713-23
references.pmid: 10089886
references.citation: Nat Biotechnol. 2004 Dec;22(12):1567-72
references.pmid: 15558047