Adeno-Associated Virus-Mediated Gene Transfer into Taste Cells In Vivo

被引:7
作者
Taruno, Akiyuki [1 ]
Kashio, Makiko [1 ]
Sun, Hongxin [1 ]
Kobayashi, Kenta [2 ,3 ]
Sano, Hiromi [3 ,4 ]
Nambu, Atsushi [2 ,3 ,4 ]
Marunaka, Yoshinori [1 ,5 ]
机构
[1] Kyoto Prefectural Univ Med, Dept Mol Cell Physiol, Kyoto 6028566, Japan
[2] Natl Inst Nat Sci, Natl Inst Physiol Sci, Sect Viral Vector Dev, Okazaki, Aichi 4448585, Japan
[3] SOKENDAI, Dept Physiol Sci, Okazaki, Aichi 4448585, Japan
[4] Natl Inst Nat Sci, Div Syst Neurophysiol, Natl Inst Physiol Sci, Okazaki, Aichi 4448585, Japan
[5] Kyoto Prefectural Univ Med, Dept Bioion, Kyoto 6028566, Japan
关键词
adeno-associated virus; AAV-DJ; lentivirus; taste; taste bud; tongue; RECEPTOR-CELLS; UMAMI TASTES; MICE LACKING; BUD CELLS; VECTOR; EXPRESSION; SWEET; PATHWAYS; RENEWAL; BITTER;
D O I
10.1093/chemse/bjw101
中图分类号
B84 [心理学]; C [社会科学总论]; Q98 [人类学];
学科分类号
03 ; 0303 ; 030303 ; 04 ; 0402 ;
摘要
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 PLC beta 2-immunoreactive type II and aromatic l-amino acid decarboxylase (AADC)-immunoreactive type III taste cells as well as in cells immunonegative for both PLC beta 2 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.
引用
收藏
页码:69 / 78
页数:10
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