Journal article
Adenovirus-mediated gene delivery to cells of the magnocellular hypothalamo-neurohypophyseal system
Experimental neurology, Vol.167(2), pp.260-271
2001
DOI: 10.1006/exnr.2000.7557
PMID: 11161614
Abstract
The objective of the present study was to define the optimum conditions for using replication-defective adenovirus (Ad) to transfer the gene for the green fluorescent protein (GFP) to the hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei and cells of the neurohypophysis (NH). As indicated by characterizing cell survival over 15 days in culture and in electrophysiological whole cell patch-clamp studies, viral concentrations up to 2 x 10(7) pfu/coverslip did not affect viability of transfected PVN and NH cultured cells from preweanling rats. At 2 x 10(7) pfu, GFP gene expression was higher (40% of GFP-positive cells) and more sustained (up to 15 days). Using a stereotaxic approach in adult rats, we were able to directly transduce the PVN, SON, and NH and visualize gene expression in coronal brain slices and in the pituitary 4 days after injection of Ad. In animals receiving NH injections of Ad, the virus was retrogradely transported to PVN and SON neurons as indicated by the appearance of GFP-positive neurons in cultures of dissociated cells from those brain nuclei and by polymerase chain reaction and Western blot analyses of PVN and SON tissues. Adenoviral concentrations of up to 8 x 10(6) pfu injected into the NH did not affect cell viability and did not cause inflammatory responses. Adenoviral injection into the pituitary enabled the selective delivery of genes to the soma of magnocellular neurons. The experimental approaches described here provide potentially useful strategies for the treatment of disordered expression of the hormones vasopressin or oxytocin.
Details
- Title: Subtitle
- Adenovirus-mediated gene delivery to cells of the magnocellular hypothalamo-neurohypophyseal system
- Creators
- Elisardo C Vasquez - 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 StatesRonald E Haskell - Departments of Psychology, Pharmacology, and Medicine and the Cardiovascular Center, University of Iowa, Iowa City, Iowa 52242, United StatesRalph F Johnson - Departments of Psychology, Pharmacology, and Medicine and the Cardiovascular Center, University of Iowa, Iowa City, Iowa 52242, United StatesSilvana S Meyrelles - Departments of Psychology, Pharmacology, and Medicine and the Cardiovascular Center, University of Iowa, Iowa City, Iowa 52242, United StatesBeverly 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.167(2), pp.260-271
- Publisher
- Elsevier
- DOI
- 10.1006/exnr.2000.7557
- PMID
- 11161614
- ISSN
- 0014-4886
- eISSN
- 1090-2430
- Grant note
- DOI: 10.13039/100000002, name: National Institutes of Health, award: DK54759, HL14388, HL57472; DOI: 10.13039/100000006, name: Office of Naval Research, award: N00014-97-1-0145; DOI: 10.13039/100000104, name: National Aeronautics and Space Administration, award: NAG5-6171; DOI: 10.13039/501100003593, name: Conselho Nacional de Desenvolvimento Científico e Tecnológico
- Language
- English
- Date published
- 2001
- Academic Unit
- Psychological and Brain Sciences; Neuroscience and Pharmacology; Health and Human Physiology
- Record Identifier
- 9984213270002771
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