An adeno-associated viral vector transduces the rat hypothalamus and amygdala more efficient than a lentiviral vector.
BACKGROUND
This study compared the transduction efficiencies of an adeno-associated viral (AAV) vector, which was pseudotyped with an AAV1 capsid and encoded the green fluorescent protein (GFP), with a lentiviral (LV) vector, which was pseudotyped with a VSV-G envelop and encoded the discosoma red fluorescent protein (dsRed), to investigate which viral vector transduced the lateral hypothalamus or the amygdala more efficiently. The LV-dsRed and AAV1-GFP vector were mixed and injected into the lateral hypothalamus or into the amygdala of adult rats. The titers that were injected were 1 x 108 or 1 x 109 genomic copies of AAV1-GFP and 1 x 105 transducing units of LV-dsRed.


RESULTS
Immunostaining for GFP and dsRed showed that AAV1-GFP transduced significantly more cells than LV-dsRed in both the lateral hypothalamus and the amygdala. In addition, the number of LV particles that were injected can not easily be increased, while the number of AAV1 particles can be increased easily with a factor 100 to 1000. Both viral vectors appear to predominantly transduce neurons.


CONCLUSIONS
This study showed that AAV1 vectors are better tools to overexpress or knockdown genes in the lateral hypothalamus and amygdala of adult rats, since more cells can be transduced with AAV1 than with LV vectors and the titer of AAV1 vectors can easily be increased to transduce the area of interest.
de Backer
Marijke W A
MW
Department of Neuroscience and Pharmacology, Rudolf Magnus Institute of Neuroscience, University Medical Centre Utrecht, Utrecht, the Netherlands.
Fitzsimons
Carlos P
CP
Brans
Maike A D
MA
Luijendijk
Mieneke C M
MC
Garner
Keith M
KM
Vreugdenhil
Erno
E
Adan
Roger A H
RA
D000679:Amygdala / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007031:Hypothalamus / Q000378:metabolism*; D007150:Immunohistochemistry; D016086:Lentivirus / Q000235:genetics* / Q000378:metabolism; D008297:Male; D009474:Neurons / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D014161:Transduction, Genetic / Q000379:methods*
D049452:Green Fluorescent Proteins