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Neural Development

, 3:22

First Online: 11 September 2008Received: 22 May 2008Accepted: 11 September 2008

Abstract

BackgroundOlfactory 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.

ResultsWe 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.

ConclusionThis 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.

AbbreviationsACadenylyl cyclase

CNGA2cyclic nucleotide gated channel A2 subunit

DIVdays in vitro

Eembryonic day

EGFPenhanced green fluorescent protein

NCAMneuronal cell adhesion molecule

OEolfactory neuroepithelia

OMPolfactory marker protein

ORodorant receptor

OSNolfactory sensory neuron

Ppostnatal day

SEstandard error

Electronic supplementary materialThe online version of this article doi:10.1186-1749-8104-3-22 contains supplementary material, which is available to authorized users.

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Autor: Huaiyang Chen - Sepehr Dadsetan - Alla F Fomina - Qizhi Gong

Fuente: https://link.springer.com/







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