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Viral gene transfer combined with bioluminescence imaging (BLI) for longitudinal monitoring of gene expression in targeted brain regions of mice
Author(s) -
Peterson J R,
Infanger D W,
Kutschke W,
Sharma R V,
Davisson R L
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.lb38-a
Subject(s) - bioluminescence imaging , luciferase , genetic enhancement , gene expression , medicine , in vivo , bioluminescence , gene , context (archaeology) , gene transfer , microbiology and biotechnology , gene delivery , pathology , biology , transfection , genetics , biochemistry , paleontology
BLI has emerged as a powerful tool for in vivo analysis of many biological processes. Our goal is to utilize this technology to map spatiotemporal patterns of transcription factor activity in CNS circuits of mice with cardiovascular disease. However, little is known about properties of bioluminescence signals (BLS) in brain, especially in the context of adenoviral (Ad) gene transfer of luciferase (luc). Here we sought to establish the short‐term kinetics and long‐term stability of the BLS following Ad‐mediated delivery of luc to mouse brain. Adult C57 mice underwent intracerebroventricular (ICV) injection (250nl) of Ad encoding the luc gene downstream of either the CMV promoter (3x10 10 pfu/ml, n=3) or NFκB response elements (1x10 11 pfu/ml, n=5). On days 2, 8 and 12 after gene transfer, serial images were taken intermittently over 24 hr following ip injection of luciferin (150 mg/kg) using the Xenogen IVIS ® 200. In mice expressing luc in the brain under either promoter, the BLS rose sharply after luciferin administration, peaked at 15 minutes, and decayed 90% by 2 hr, returning to baseline by 24 hr. The peak BLS was robust (eg day 8 AdCMVluc 5.3±.6x10 5 photons/s) and remained stable out to day 12 (eg 5.6±.5x10 5 photons/s). Planar spectral analysis revealed a BLS source depth of 1.9±.2mm (n=5) from the skull surface, coinciding with the stereotaxic location of brain nuclei known to be targeted by ICV viral gene delivery. This demonstrates the utility of BLI and establishes optimal parameters for serial monitoring of spatiotemporal patterns of gene expression in targeted brain regions of living mice.