PMID- 21431420
OWN - NLM
STAT- MEDLINE
DCOM- 20111130
LR  - 20211020
IS  - 1573-6830 (Electronic)
IS  - 0272-4340 (Print)
IS  - 0272-4340 (Linking)
VI  - 31
IP  - 6
DP  - 2011 Aug
TI  - Targeting neurons of rat nucleus tractus solitarii with the gene transfer vector 
      adeno-associated virus type 2 to up-regulate neuronal nitric oxide synthase.
PG  - 847-59
LID - 10.1007/s10571-011-9674-5 [doi]
AB  - Adeno-associated virus (AAV) has distinct advantages over other viral vectors in 
      delivering genes of interest to the brain. AAV mainly transfects neurons, 
      produces no toxicity or inflammatory responses, and yields long-term transgene 
      expression. In this study, we first tested the hypothesis that AAV serotype 2 
      (AAV2) selectively transfects neurons but not glial cells in the nucleus tractus 
      solitarii (NTS) by examining expression of the reporter gene, enhanced green 
      fluorescent protein (eGFP), in the rat NTS after unilateral microinjection of 
      AAV2eGFP into NTS. Expression of eGFP was observed in 1-2 cells in the NTS 1 day 
      after injection. The number of transduced cells and the intensity of eGFP 
      fluorescence increased from day 1 to day 28 and decreased on day 60. The majority 
      (92.9 +/- 7.0%) of eGFP expressing NTS cells contained immunoreactivity for the 
      neuronal marker, protein gene product 9.5, but not that for the glial marker, 
      glial fibrillary acidic protein. We observed eGFP expressing neurons and fibers 
      in the nodose ganglia (NG) both ipsilateral and contralateral to the injection. 
      In addition, eGFP expressing fibers were present in both ipsilateral and 
      contralateral nucleus ambiguus (NA), caudal ventrolateral medulla (CVLM) and 
      rostral ventrolateral medulla (RVLM). Having established that AAV2 was able to 
      transduce a gene into NTS neurons, we constructed AAV2 vectors that contained 
      cDNA for neuronal nitric oxide synthase (nNOS) and examined nNOS expression in 
      the rat NTS after injection of this vector into the area. Results from RT-PCR, 
      Western analysis, and immunofluorescent histochemistry indicated that nNOS 
      expression was elevated in rat NTS that had been injected with AAV2nNOS vectors. 
      Therefore, we conclude that AAV2 is an effective viral vector in chronically 
      transducing NTS neurons and that AAV2nNOS can be used as a specific gene transfer 
      tool to study the role of nNOS in CNS neurons.
FAU - Lin, Li-Hsien
AU  - Lin LH
AD  - Department of Neurology, University of Iowa, Iowa City, IA 52242, USA. 
      li-hsien-lin@uiowa.edu
FAU - Dragon, Deidre Nitschke
AU  - Dragon DN
FAU - Jin, Jingwen
AU  - Jin J
FAU - Talman, William T
AU  - Talman WT
LA  - eng
GR  - R01 HL088090/HL/NHLBI NIH HHS/United States
GR  - R01 HL059593/HL/NHLBI NIH HHS/United States
GR  - R01 HL 088090/HL/NHLBI NIH HHS/United States
GR  - R01 HL 59593/HL/NHLBI NIH HHS/United States
GR  - R01 HL088090-04/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20110323
PL  - United States
TA  - Cell Mol Neurobiol
JT  - Cellular and molecular neurobiology
JID - 8200709
RN  - 0 (enhanced green fluorescent protein)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - EC 1.14.13.39 (Nitric Oxide Synthase Type I)
SB  - IM
MH  - Animals
MH  - Blotting, Western
MH  - Dependovirus/*genetics
MH  - *Gene Transfer Techniques
MH  - Genetic Vectors/*genetics
MH  - Green Fluorescent Proteins/metabolism
MH  - HEK293 Cells
MH  - Humans
MH  - Injections
MH  - Male
MH  - Medulla Oblongata/cytology/metabolism
MH  - Microscopy, Confocal
MH  - Neurons/cytology/*metabolism
MH  - Nitric Oxide Synthase Type I/*genetics/metabolism
MH  - Nodose Ganglion/cytology/metabolism
MH  - Organ Specificity
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Solitary Nucleus/*cytology/metabolism
MH  - Time Factors
MH  - Transduction, Genetic
MH  - Up-Regulation/*genetics
PMC - PMC3146586
MID - NIHMS301391
EDAT- 2011/03/25 06:00
MHDA- 2011/12/13 00:00
PMCR- 2011/08/01
CRDT- 2011/03/25 06:00
PHST- 2011/01/27 00:00 [received]
PHST- 2011/03/05 00:00 [accepted]
PHST- 2011/03/25 06:00 [entrez]
PHST- 2011/03/25 06:00 [pubmed]
PHST- 2011/12/13 00:00 [medline]
PHST- 2011/08/01 00:00 [pmc-release]
AID - 10.1007/s10571-011-9674-5 [doi]
PST - ppublish
SO  - Cell Mol Neurobiol. 2011 Aug;31(6):847-59. doi: 10.1007/s10571-011-9674-5. Epub 
      2011 Mar 23.