title: Quantification of Cerebral Vascular Architecture using Two-photon Microscopy in a Mouse Model of HIV-induced Neuroinflammation.
abstract: Human Immunodeficiency Virus 1 (HIV-1) infection frequently results in HIV-1 Associated Neurocognitive Disorders (HAND), and is characterized by a chronic neuroinflammatory state within the central nervous system (CNS), thought to be driven principally by virally-mediated activation of microglia and brain resident macrophages. HIV-1 infection is also accompanied by changes in cerebrovascular blood flow (CBF), raising the possibility that HIV-associated chronic neuroinflammation may lead to changes in CBF and/or in cerebral vascular architecture. To address this question, we have used a mouse model for HIV-induced neuroinflammation, and we have tested whether long-term exposure to this inflammatory environment may damage brain vasculature and result in rarefaction of capillary networks. In this paper we describe a method to quantify changes in cortical capillary density in a mouse model of neuroinflammatory disease (HIV-1 Tat transgenic mice). This generalizable approach employs in vivo two-photon imaging of cortical capillaries through a thin-skull cortical window, as well as ex vivo two-photon imaging of cortical capillaries in mouse brain sections. These procedures produce images and z-stack files of capillary networks, respectively, which can be then subjected to quantitative analysis in order to assess changes in cerebral vascular architecture.
authors: Department of Microbiology and Immunology, University of Rochester Medical Center.
authors: Christopher
authors: C
authors: Nishimura
authors: Department of Human Genetics, University of Chicago.
authors: Oksana
authors: O
authors: Polesskaya
authors: Department of Microbiology and Immunology, University of Rochester Medical Center; Stephen_Dewhurst@urmc.rochester.edu.
authors: Stephen
authors: S
authors: Dewhurst
authors: Department of Microbiology and Immunology, University of Rochester Medical Center.
authors: Jharon N
authors: JN
authors: Silva
mesh_terms: D000818:Animals; D001921:Brain / Q000098:blood supply* / Q000473:pathology; D002560:Cerebrovascular Circulation; D004195:Disease Models, Animal; D005456:Fluorescent Dyes / Q000737:chemistry; D015658:HIV Infections / Q000473:pathology*; D015497:HIV-1; D006801:Humans; D007249:Inflammation / Q000473:pathology / Q000821:virology; D051379:Mice; D008822:Mice, Transgenic; D017628:Microglia / Q000473:pathology; D008856:Microscopy, Fluorescence / Q000379:methods; D017785:Photons; D012886:Skull / Q000601:surgery
chemical_list: D005456:Fluorescent Dyes