Regulated delivery of glial cell line-derived neurotrophic factor into rat striatum, using a tetracycline-dependent lentiviral vector.
Georgievska, Biljana LU ; Jakobsson, Johan LU
; Persson, Elisabeth
; Ericson, Cecilia
LU
; Kirik, Deniz
LU
and Lundberg, Cecilia
LU
(2004)
In Human Gene Therapy
15(10).
p.934-944
- Abstract
- In this study, a tetracycline-regulated lentiviral vector system, based on the tetracycline-dependent transactivator rtTA2S-M2, was developed for controlled expression of glial cell line-derived neurotrophic factor (GDNF) in the rat brain. Expression of the marker gene green fluorescent protein (GFP) and GDNF was tightly regulated in a dose-dependent manner in neural cell lines in vitro. Injection of high-titer lentiviral vectors into the rat striatum resulted in a 7-fold induction of GDNF tissue levels (1060 pg/mg tissue), when doxycycline (a tetracycline analog) was added to the drinking water. However, low levels of GDNF (150 pg/mg tissue) were also detected in animals that did not receive doxycycline, indicating a significant... (More)
- In this study, a tetracycline-regulated lentiviral vector system, based on the tetracycline-dependent transactivator rtTA2S-M2, was developed for controlled expression of glial cell line-derived neurotrophic factor (GDNF) in the rat brain. Expression of the marker gene green fluorescent protein (GFP) and GDNF was tightly regulated in a dose-dependent manner in neural cell lines in vitro. Injection of high-titer lentiviral vectors into the rat striatum resulted in a 7-fold induction of GDNF tissue levels (1060 pg/mg tissue), when doxycycline (a tetracycline analog) was added to the drinking water. However, low levels of GDNF (150 pg/mg tissue) were also detected in animals that did not receive doxycycline, indicating a significant background leakage from the vector system in vivo. The level of basal expression was markedly reduced when a 10-fold lower dose of the tetracycline-regulated GDNF vector was injected into the striatum (3–11 pg/mg tissue), and doxycycline- induced GDNF tissue levels obtained in these animals were about 190 pg/mg tissue. Doxycycline-induced expression of GDNF resulted in a significant downregulation of the tyrosine hydroxylase (TH) protein in the intact striatum. Removal of doxycycline from the drinking water rapidly (within 3 days) turned off transgenic GDNF mRNA expression and GDNF protein levels in the tissue were completely reduced by 2 weeks, demonstrating the dynamics of the system in vivo. Accordingly, TH protein expression returned to normal by 2–8 weeks after removal of doxycycline, indicating that GDNF-induced downregulation of TH is a reversible event. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/132118
- author
- Georgievska, Biljana
LU
; Jakobsson, Johan
LU
; Persson, Elisabeth
; Ericson, Cecilia
LU
; Kirik, Deniz
LU
and Lundberg, Cecilia
LU
- organization
- publishing date
- 2004
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Human Gene Therapy
- volume
- 15
- issue
- 10
- pages
- 934 - 944
- publisher
- Mary Ann Liebert, Inc.
- external identifiers
-
- wos:000224578600002
- pmid:15585109
- scopus:5644254312
- ISSN
- 1043-0342
- DOI
- 10.1089/hum.2004.15.934
- language
- English
- LU publication?
- yes
- id
- 792be8ea-6083-4bc3-b40c-13c29acb81c9 (old id 132118)
- date added to LUP
- 2016-04-01 15:41:58
- date last changed
- 2022-01-28 06:38:30
@article{792be8ea-6083-4bc3-b40c-13c29acb81c9,
abstract = {{In this study, a tetracycline-regulated lentiviral vector system, based on the tetracycline-dependent transactivator rtTA2S-M2, was developed for controlled expression of glial cell line-derived neurotrophic factor (GDNF) in the rat brain. Expression of the marker gene green fluorescent protein (GFP) and GDNF was tightly regulated in a dose-dependent manner in neural cell lines in vitro. Injection of high-titer lentiviral vectors into the rat striatum resulted in a 7-fold induction of GDNF tissue levels (1060 pg/mg tissue), when doxycycline (a tetracycline analog) was added to the drinking water. However, low levels of GDNF (150 pg/mg tissue) were also detected in animals that did not receive doxycycline, indicating a significant background leakage from the vector system in vivo. The level of basal expression was markedly reduced when a 10-fold lower dose of the tetracycline-regulated GDNF vector was injected into the striatum (3–11 pg/mg tissue), and doxycycline- induced GDNF tissue levels obtained in these animals were about 190 pg/mg tissue. Doxycycline-induced expression of GDNF resulted in a significant downregulation of the tyrosine hydroxylase (TH) protein in the intact striatum. Removal of doxycycline from the drinking water rapidly (within 3 days) turned off transgenic GDNF mRNA expression and GDNF protein levels in the tissue were completely reduced by 2 weeks, demonstrating the dynamics of the system in vivo. Accordingly, TH protein expression returned to normal by 2–8 weeks after removal of doxycycline, indicating that GDNF-induced downregulation of TH is a reversible event.}},
author = {{Georgievska, Biljana and Jakobsson, Johan and Persson, Elisabeth and Ericson, Cecilia and Kirik, Deniz and Lundberg, Cecilia}},
issn = {{1043-0342}},
language = {{eng}},
number = {{10}},
pages = {{934--944}},
publisher = {{Mary Ann Liebert, Inc.}},
series = {{Human Gene Therapy}},
title = {{Regulated delivery of glial cell line-derived neurotrophic factor into rat striatum, using a tetracycline-dependent lentiviral vector.}},
url = {{http://dx.doi.org/10.1089/hum.2004.15.934}},
doi = {{10.1089/hum.2004.15.934}},
volume = {{15}},
year = {{2004}},
}