Expressing exogenous functional odorant receptors in cultured olfactory sensory neurons

Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis, California 95616, USA.
Neural Development (Impact Factor: 3.45). 10/2008; 3(1):22. DOI: 10.1186/1749-8104-3-22
Source: PubMed


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.
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.
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.

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    • "To produce functional ORs, some groups have used various OR expression systems, either mammalian cells (Chen, Dadsetan, Fomina, & Gong, 2008; Dahoun, Grasso, Vogel, & Pick, 2011; Ko & Park, 2006; Krautwurst, Yau, & Reed, 1998; Levasseur et al., 2003; Matsunami, Mainland, & Dey, 2009; Saito, Kubota, Roberts, Chi, & Matsunami, 2004; Zhuang & Matsunami, 2007), Saccharomyces cerevisiae-based cells (Fukutani et al., 2012; Minic et al., 2005; Pajot-Augy, Crowe, Levasseur, Salesse, & Connerton, 2003; Radhika et al., 2007) or Escherichia coli cells (Song, Lee, Oh, & Park, 2009; Sung, Ko, & Park, 2006) also regarded as most promising systems. Here, ORs are produced in S. cerevisiae, which constitutes an efficient heterologous expression system. "
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