title: Non-invasive detection of spatio-temporal activation of SBE and NFAT5 promoters in transgenic reporter mice following stroke.
abstract: The characterization of molecular responses following cerebral ischemia-induced changes in animal models capable of undergoing real-time analysis is an important goal for stroke research. In this study, we use transgenic mice to examine the activation of two different promoters in a firefly luciferase reporter mouse analyzable through a non-invasive bioluminescent imaging system. In the first model, we examine the middle cerebral artery occlusion (MCAO)-induced activation of Smad-binding elements (SBE), a downstream target of Smad 1/2/3 transcription factors, in which SBEs regulate the expression of the fluc reporter. We observed that MCAO induces a bilateral activation (i.e., both ipsilateral and contralateral brain hemispheres) of the SBE-luc reporter with a peak at 24 h. In the second model, we examined MCAO-induced activation of the osmolarity-sensitive promoter nuclear factor of activated T-cell 5 (NFAT5) and identified a peak reporter expression 72 h post-MCAO in the ipsilateral but not contralateral hemisphere. In each of these models, the assessment of post-MCAO fluc-expression provided both a quantitative measure (i.e., radiance in photons/sec/cm(2) /steradian) as well as qualitative localization of the molecular response following focal ischemic injury.
authors.affiliation: Department of Surgery, University of California of San Diego, San Diego, California 92121, USA. beliceiri@ucsd.edu
authors.forename: Ashkaun
authors.initials: A
authors.lastname: Shaterian
authors.forename: Alexandra
authors.initials: A
authors.lastname: Borboa
authors.forename: Raul
authors.initials: R
authors.lastname: Coimbra
authors.forename: Andrew
authors.initials: A
authors.lastname: Baird
authors.forename: Brian P
authors.initials: BP
authors.lastname: Eliceiri
mesh_terms: D000818:Animals; D017930:Genes, Reporter / Q000502:physiology*; D008156:Luciferases / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D051899:Smad2 Protein / Q000235:genetics* / Q000378:metabolism; D051900:Smad3 Protein / Q000235:genetics* / Q000378:metabolism; D020521:Stroke / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
chemical_list: C492200:Nfat5 protein, mouse; D051899:Smad2 Protein; C497783:Smad2 protein, mouse; D051900:Smad3 Protein; C497791:Smad3 protein, mouse; D014157:Transcription Factors; D008156:Luciferases