Distinct patterns of gene transfer to gerbil hippocampus with recombinant adeno-associated virus type 2 and 5.
Genetic modification of the gerbil hippocampal neuronal cells in vivo helps us understand the mechanisms of neuronal function under various circumstances such as ischemic insult. In this study, we examined the distinct distribution of the recombinant adeno-associated virus type 2 (rAAV2) and rAAV5 vectors for gene delivery to primary cultured cells and the gerbil hippocampus. Mixed cortical cultures containing both neurons and astrocytes from E17 rat embryos were infected with rAAVs containing the Cytomegalovirus virus (CMV) promoter. rAAV2 was preferably transduced to neurons, whereas rAAV5 was inclined to be transduced to astrocytes in vitro. rAAV2 and rAAV5 vectors, each with the CMV or Rous sarcoma virus (RSV) promoter, were injected into the gerbil hippocampus using a stereotaxic apparatus. Five days after injection, transgene expression was analyzed with X-gal staining. In the gerbil hippocampus, rAAV5 with the CMV promoter achieved a higher overall transgene expression than rAAV2 with the CMV promoter. The transgene expression of rAAV2 with the RSV promoter was found in the pyramidal and granular cells, while the transgene expression of rAAV5 with the RSV promoter was preferentially found in the granular cells. These findings would be valuable in optimizing rAAV-mediated gene transfer to the gerbil hippocampus.
Nomoto
Tatsuya
T
Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical School 3311-1 Yakushiji, Minami-kawachi, Kawachi, Tochigi 329-0498, Japan.
Okada
Takashi
T
Shimazaki
Kuniko
K
Mizukami
Hiroaki
H
Matsushita
Takashi
T
Hanazono
Yutaka
Y
Kume
Akihiro
A
Katsura
Ken-ichiro
K
Katayama
Yasuo
Y
Ozawa
Keiya
K
D000818:Animals; D030801:Animals, Genetically Modified; D002478:Cells, Cultured; D000229:Dependovirus / Q000235:genetics*; D016503:Drug Delivery Systems / Q000379:methods; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D005849:Gerbillinae; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D019076:Transgenes / Q000235:genetics*