Open AccessModel System for Live Imaging of Neuronal Responses to Injury and Repair
Author(s) -
Mathieu Gravel,
Yuan-Cheng Weng,
Jasna Križ
Publication year - 2011
Publication title -
molecular imaging
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.815
H-Index - 60
eISSN - 1536-0121
pISSN - 1535-3508
DOI - 10.2310/7290.2011.00013
Subject(s) - colocalization , green fluorescent protein , neurite , bioluminescence , bioluminescence imaging , biology , microbiology and biotechnology , genetically modified mouse , transgene , reporter gene , gap junction , gap 43 protein , endogeny , neuroscience , luciferase , gene expression , cell culture , immunohistochemistry , transfection , gene , immunology , in vitro , endocrinology , ecology , biochemistry , genetics , intracellular
Although it has been well established that induction of growth-associated protein-43 (GAP-43) during development coincides with axonal outgrowth and early synapse formation, the existence of neuronal plasticity and neurite outgrowth in the adult central nervous system after injuries is more controversial. To visualize the processes of neuronal injury and repair in living animals, we generated reporter mice for bioluminescence and fluorescence imaging bearing the luc (luciferase) and gfp (green fluorescent protein) reporter genes under the control of the murine GAP-43 promoter. Reporter functionality was first observed during the development of transgenic embryos. Using in vivo bioluminescence and fluorescence imaging, we visualized induction of the GAP-43 signals from live embryos starting at E10.5, as well as neuronal responses to brain and peripheral nerve injuries (the signals peaked at 14 days postinjury). Moreover, three-dimensional analysis of the GAP-43 bioluminescent signal confirmed that it originated from brain structures affected by ischemic injury. The analysis of fluorescence signal at cellular level revealed colocalization between endogenous protein and the GAP-43-driven gfp transgene. Taken together, our results suggest that the GAP-43-luc/gfp reporter mouse represents a valid model system for real-time analysis of neurite outgrowth and the capacity of the adult nervous system to regenerate after injuries