title: Adeno-Associated Virus-Mediated Gene Transfer into Taste Cells In Vivo.
abstract: The sense of taste is achieved by cooperation of many signaling molecules expressed in taste cells, which code and transmit information on quality and intensity of taste to the nervous system. Viral vector-mediated gene transfer techniques have been proven to be useful to study and control function of a gene product in vivo However, there is no transduction method for taste cells in live animals. Here, we have established a method for inducing foreign gene expression in mouse taste cells in vivo by recombinant adeno-associated virus (AAV) vector. First, using enhanced green fluorescent protein (EGFP) as a reporter, we screened 6 AAV serotypes along with a recombinant lentivirus vector for their ability to transduce taste cells. One week after viral injection into the submucosa of the tongue, EGFP expression in fungiform taste cells was observed only in animals injected with AAV-DJ, a synthetic serotype. Next, time course of AAV-DJ-mediated EGFP expression in fungiform taste cells was evaluated. Intragemmal EGFP signals appeared after a delay, rapidly increased until 7 days postinjection, and gradually decreased over the next few weeks probably because of the cell turnover. Finally, the taste cell types susceptible to AAV-DJ transduction were characterized. EGFP expression was observed in PLCβ2-immunoreactive type II and aromatic l-amino acid decarboxylase (AADC)-immunoreactive type III taste cells as well as in cells immunonegative for both PLCβ2 and AADC, demonstrating that AAV-DJ does not discriminate functional taste cell types. In conclusion, the method established in this study will be a promising tool to study the mechanism of taste.
authors.affiliation: Department of Molecular Cell Physiology, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan.
authors.forename: Akiyuki
authors.initials: A
authors.lastname: Taruno
authors.affiliation: Department of Molecular Cell Physiology, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan.
authors.forename: Makiko
authors.initials: M
authors.lastname: Kashio
authors.affiliation: Department of Molecular Cell Physiology, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan.
authors.forename: Hongxin
authors.initials: H
authors.lastname: Sun
authors.affiliation: Section of Viral Vector Development, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki 444-8585, Japan.|Department of Physiological Sciences, SOKENDAI (The Graduate University for Advanced Studies), Okazaki 444-8585, Japan.
authors.forename: Kenta
authors.initials: K
authors.lastname: Kobayashi
authors.affiliation: Department of Physiological Sciences, SOKENDAI (The Graduate University for Advanced Studies), Okazaki 444-8585, Japan.|Division of System Neurophysiology, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki 444-8585, Japan and.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Sano
authors.affiliation: Section of Viral Vector Development, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki 444-8585, Japan.|Department of Physiological Sciences, SOKENDAI (The Graduate University for Advanced Studies), Okazaki 444-8585, Japan.|Division of System Neurophysiology, National Institute for Physiological Sciences, National Institutes of Natural Sciences, Okazaki 444-8585, Japan and.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Nambu
authors.affiliation: Department of Molecular Cell Physiology, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan.|Department of Bio-Ionomics, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan.
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Marunaka
mesh_terms: D000818:Animals; D000229:Dependovirus / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D013650:Taste Buds / Q000166:cytology* / Q000378:metabolism*
keywords: AAV-DJ; adeno-associated virus; lentivirus; taste; taste bud; tongue