Journal article
Replication-deficient adenovirus vector transfer of gfp reporter gene into supraoptic nucleus and subfornical organ neurons
Experimental neurology, Vol.154(2), pp.353-365
1998
DOI: 10.1006/exnr.1998.6917
PMID: 9878173
Abstract
The present studies used defined cells of the subfornical organ (SFO) and supraoptic nuclei (SON) as model systems to demonstrate the efficacy of replication-deficient adenovirus (Ad) encoding green fluorescent protein (GFP) for gene transfer. The studies investigated the effects of both direct transfection of the SON and indirect transfection (i.e., via retrograde transport) of SFO neurons. The SON of rats were injected with Ad (2 × 106pfu) and sacrificed 1–7 days later for cell culture of the SON and of the SFO. In the SON, GFP fluorescence was visualized in both neuronal and nonneuronal cells while only neurons in the SFO expressed GFP. Successfulin vitrotransfection of cultured cells from the SON and SFO was also achieved with Ad (2 × 106to 2 × 108pfu). The expression of GFP inin vitrotransfected cells was higher in nonneuronal (approximately 28% in SON and SFO) than neuronal (approximately 4% in SON and 10% in SFO) cells. The expression of GFP was time and viral concentration related. No apparent alterations in cellular morphology of transfected cells were detected and electrophysiological characterization of transfected cells was similar between GFP-expressing and nonexpressing neurons. We conclude that (1) GFP is an effective marker for gene transfer in living SON and SFO cells, (2) Ad infects both neuronal and nonneuronal cells, (3) Ad is taken up by axonal projections from the SON and retrogradely transported to the SFO where it is expressed at detectable levels, and (4) Ad does not adversely affect neuronal viability. These results demonstrate the feasibility of using adenoviral vectors to deliver genes to the SFO–SON axis.
Details
- Title: Subtitle
- Replication-deficient adenovirus vector transfer of gfp reporter gene into supraoptic nucleus and subfornical organ neurons
- Creators
- Elisardo C Vasquez - Departments of Psychology, Pharmacology and Medicine and the Cardiovascular Center University of Iowa, Iowa City, Iowa 52242, United StatesR. F Johnson - Departments of Psychology, Pharmacology and Medicine and the Cardiovascular Center University of Iowa, Iowa City, Iowa 52242, United StatesTerry G Beltz - Departments of Psychology, Pharmacology and Medicine and the Cardiovascular Center University of Iowa, Iowa City, Iowa 52242, United StatesR. E Haskell - Departments of Psychology, Pharmacology and Medicine and the Cardiovascular Center University of Iowa, Iowa City, Iowa 52242, United StatesB. L Davidson - Departments of Psychology, Pharmacology and Medicine and the Cardiovascular Center University of Iowa, Iowa City, Iowa 52242, United StatesAlan Kim Johnson - Departments of Psychology, Pharmacology and Medicine and the Cardiovascular Center University of Iowa, Iowa City, Iowa 52242, United States
- Resource Type
- Journal article
- Publication Details
- Experimental neurology, Vol.154(2), pp.353-365
- Publisher
- Elsevier
- DOI
- 10.1006/exnr.1998.6917
- PMID
- 9878173
- ISSN
- 0014-4886
- eISSN
- 1090-2430
- Language
- English
- Date published
- 1998
- Academic Unit
- Psychological and Brain Sciences; Neuroscience and Pharmacology; Health and Human Physiology
- Record Identifier
- 9984213276302771
Metrics
3 Record Views