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Record 20 of 260
Adenovirus-mediated gene delivery to cells of the magnocellular hypothalamo-neurohypophyseal system
External Online Source: doi:10.1006/exnr.2000.7557
Author and Affiliation:
Vasquez, E. C.(University of Iowa, Department of Psychology, Iowa City, Iowa 52242, United States)
Beltz, T. G.
Haskell, R. E.
Johnson, R. F.
Meyrelles, S. S.
Davidson, B. L.
Johnson, A. K.
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. Copyright 2000 Academic Press.
Publication Date: Feb 01, 2001
Document ID:
20040112601
(Acquired Oct 05, 2004)
Subject Category: LIFE SCIENCES (GENERAL)
Document Type: Journal Article
Publication Information: Experimental neurology (ISSN 0014-4886); Volume 167; 2; 260-71
Publisher Information: United States
Contract/Grant/Task Num: DK54759; HL04388; HL57472
Description: In English
Distribution Limits: Unclassified; Publicly available; Unlimited
Rights: Copyright
NASA Terms: ADENOVIRUSES; GENES; HYPOTHALAMUS; METABOLISM; BIOSYNTHESIS; CULTURED CELLS; CYTOLOGY; FEMALES; GENE EXPRESSION; GENETICS; LUMINESCENT PROTEINS; MALES; PITUITARY GLAND; RATS; SURGERY
Other Descriptors: ADENOVIRIDAE/GENETICS/METABOLISM; GENE TRANSFER TECHNIQUES; HYPOTHALAMO-HYPOPHYSEAL SYSTEM/CYTOLOGY/METABOLISM/SURGERY; ANIMALS; CELLS, CULTURED; FEMALE; GENE EXPRESSION; LUMINESCENT PROTEINS/BIOSYNTHESIS/GENETICS; MALE; PARAVENTRICULAR HYPOTHALAMIC NUCLEUS/CYTOLOGY/METABOLISM/VIROLOGY; PATCH-CLAMP TECHNIQUES; PITUITARY GLAND, POSTERIOR/CYTOLOGY/METABOLISM/SURGERY/VIROLOGY; RATS; RATS, SPRAGUE-DAWLEY; SUPPORT, NON-U.S. GOV'T; SUPPORT, U.S. GOV'T, NON-P.H.S; SUPPORT, U.S. GOV'T, P.H.S; SUPRAOPTIC NUCLEUS/CYTOLOGY/METABOLISM/VIROLOGY; TRANSFECTION; NASA DISCIPLINE REGULATORY PHYSIOLOGY; NON-NASA CENTER
Availability Source: Other Sources
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