title: Adenovirus-mediated transfer and expression of beta-gal in injured hippocampus after traumatic brain injury in mice.
abstract: In models of focal cerebral ischemia, adenoviral gene transfer is often attenuated or delayed versus naive. After controlled cortical impact (CCI)-induced traumatic brain injury in mice, CA1 and CA3 hippocampus exhibit delayed neuronal death by 3 days, with subsequent near complete loss of hippocampus by 21 days. We hypothesized that adenoviral-mediated expression of the reporter gene beta-Galactosidase (beta-Gal) in hippocampus would be attenuated after CCI in mice. C57BL6 mice (n = 16) were subjected to either CCI to left parietal cortex or sham (burr hole). Adenovirus carrying the beta-Gal gene (AdlacZ; 1 x 10(9) plaque-forming units [pfu]/mL) was then injected into left dorsal hippocampus. At 24 or 72 h, beta-Gal expression was quantified (mU/mg protein). Separate mice (n = 10) were used to study beta-Gal spatial distribution in brain sections. Beta-Gal expression in left hippocampus was similar in shams at 24 h (48.4 +/- 4.1) versus 72 h (68.8 +/- 8.8, not significant). CCI did not reduce beta-Gal expression in left hippocampus (68.8 +/- 8.8 versus 88.1 +/- 7.0 at 72 h, sham versus CCI, not significant). In contrast, CCI reduced beta-Gal expression in right (contralateral) hippocampus versus sham (p < 0.05 at both 24 and 72 h). Beta-Gal was seen in many cell types in ipsilateral hippocampus, including CA3 neurons. Despite eventual loss of ipsilateral hippocampus, adenovirus-mediated gene transfer was surprisingly robust early after CCI providing an opportunity to test novel genes targeting delayed hippocampal neuronal death.
authors.affiliation: Safar Center for Resuscitation Research, Pittsburgh, PA 15260, USA. kochanekpm@anes.upmc.edu
authors.forename: P M
authors.initials: PM
authors.lastname: Kochanek
authors.forename: K L
authors.initials: KL
authors.lastname: Janesko
authors.forename: L W
authors.initials: LW
authors.lastname: Jenkins
authors.forename: H Q
authors.initials: HQ
authors.lastname: Yan
authors.forename: M R
authors.initials: MR
authors.lastname: Kibbe
authors.forename: P
authors.initials: P
authors.lastname: Robichaud
authors.forename: A C
authors.initials: AC
authors.lastname: Wooditch
authors.forename: R S
authors.initials: RS
authors.lastname: Clark
authors.forename: C E
authors.initials: CE
authors.lastname: Dixon
authors.forename: D W
authors.initials: DW
authors.lastname: Marion
authors.forename: T R
authors.initials: TR
authors.lastname: Billiar
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001930:Brain Injuries / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D002560:Cerebrovascular Circulation / Q000502:physiology; D004195:Disease Models, Animal; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D017930:Genes, Reporter / Q000502:physiology*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000502:physiology*; D006624:Hippocampus / Q000293:injuries* / Q000378:metabolism / Q000473:pathology; D051379:Mice; D009410:Nerve Degeneration / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control; D009414:Nerve Growth Factors / Q000096:biosynthesis / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism
chemical_list: D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D001616:beta-Galactosidase