title: Intravital imaging of axonal interactions with microglia and macrophages in a mouse dorsal column crush injury.
abstract: Traumatic spinal cord injury causes an inflammatory reaction involving blood-derived macrophages and central nervous system (CNS)-resident microglia. Intra-vital two-photon microscopy enables the study of macrophages and microglia in the spinal cord lesion in the living animal. This can be performed in adult animals with a traumatic injury to the dorsal column. Here, we describe methods for distinguishing macrophages from microglia in the CNS using an irradiation bone marrow chimera to obtain animals in which only macrophages or microglia are labeled with a genetically encoded green fluorescent protein. We also describe a injury model that crushes the dorsal column of the spinal cord, thereby producing a simple, easily accessible, rectangular lesion that is easily visualized in an animal through a laminectomy. Furthermore, we will outline procedures to sequentially image the animals at the anatomical site of injury for the study of cellular interactions during the first few days to weeks after injury.
authors.affiliation: Department of Neurosciences, Case Western Reserve University.
authors.forename: Teresa A
authors.initials: TA
authors.lastname: Evans
authors.affiliation: Department of Biomedical Engineering, Case Western Reserve University.
authors.forename: Deborah S
authors.initials: DS
authors.lastname: Barkauskas
authors.affiliation: Department of Pediatrics, Case Western Reserve University.
authors.forename: Jay T
authors.initials: JT
authors.lastname: Myers
authors.affiliation: Department of Pediatrics, Case Western Reserve University; alex.y.huang@case.edu.
authors.forename: Alex Y
authors.initials: AY
authors.lastname: Huang
mesh_terms: D000818:Animals; D001369:Axons / Q000473:pathology*; D002450:Cell Communication / Q000502:physiology*; D004195:Disease Models, Animal; D007249:Inflammation / Q000473:pathology; D000069416:Intravital Microscopy / Q000379:methods*; D008264:Macrophages / Q000473:pathology*; D051379:Mice; D017628:Microglia / Q000473:pathology*; D013119:Spinal Cord Injuries / Q000473:pathology*