Full Description
Blood-brain barrier (BBB) breakdown leading to cerebral edema occurs in many brain diseases—such as trauma, stroke, inflammation, infection, and tumors—and is an important factor in the mortality arising from these con- tions. Despite the importance of the BBB in the pathogenesis of these diseases, the molecular mechanisms occurring at the BBB are not completely und- stood. In the last decade a number of molecules have been identified not only in endothelial cells, but also in astrocytes, pericytes, and the perivascular cells that interact with endothelium to maintain cerebral homeostasis. However, the precise cellular interactions at a molecular level in steady states and d- eases have still to be determined. The introduction of new research techniques during the last decade or so provide an opportunity to study the molecular mec- nisms occurring at the BBB in diseases. The Blood-Brain Barrier: Biology and Research Protocols provides the reader with details of selected morphologic, permeability, transport, in vitro, and molecular techniques for BBB studies, all written by experts in the field. Each part is preceded by a review that emphasizes the advantages and pitfalls of particular techniques, as well as offering much relevant current information. The techniques provided will be helpful to both beginners in BBB research and those more experienced investigators who wish to add a specific technique to those already available in their laboratories.
Contents
Tissue Techniques.- Morphology and Molecular Properties of Cellular Components of Normal Cerebral Vessels.- Studies of Cerebral Vessels by Transmission Electron Microscopy and Morphometry.- Freeze-Fracture Studies of Cerebral Endothelial Tight Junctions.- Studies of Cerebral Endothelium by Scanning and High-Voltage Electron Microscopy.- Detection of Endothelial/Lymphocyte Interaction in Spinal Cord Microvasculature by Intravital Videomicroscopy.- Blood-Brain Barrier Permeability Techniques.- Pathophysiology of Blood-Brain Barrier Breakdown.- Pial Vessel Permeability to Tracers Using Cranial Windows.- Blood-Brain Barrier Permeability Using Tracers and Immunohistochemistry.- Ultracytochemical Studies of the Compromised Blood-Brain Barrier.- Immunogold Detection of Microvascular Proteins in the Compromised Blood-Brain Barrier.- Measurement of Blood-Brain and Blood-Tumor Barrier Permeabilities with [14C]-Labeled Tracers.- Blood-Brain Barrier Transport Techniques.- A Review of Blood-Brain Barrier Transport Techniques.- In Situ Brain Perfusion Technique.- Brain Efflux Index Method.- Transport Studies Using Membrane Vesicles.- Drug Transport Studies Using Quantitative Microdialysis.- Single-Pass Dual-Label Indicator Method.- Protein Transport in Cerebral Endothelium.- Transport Across the Choroid Plexuses In Vivo and In Vitro.- In Vitro Techniques.- An Overview of In Vitro Techniques for Blood-Brain Barrier Studies.- Isolation and Characterization of Human Brain Endothelial Cells.- Human Brain Microvessel Endothelial Cell and Leukocyte Interactions.- Development and Characterization of Immortalized Cerebral Endothelial Cell Lines.- Isolation and Characterization of Retinal Endothelial Cells.- Isolation and Characterization of Cerebral Microvascular Pericytes.- Molecular Techniques.- Molecular Biology of the Blood-Brain Barrier.- Blood-Brain Barrier Genomics.- Dynamic In Vitro Model of the Blood-Brain Barrier.- Phage Display Technology for Identifying Specific Antigens on Brain Endothelial Cells.- mRNA Detection in Cerebral Vessels by Nonradioactive In Situ Hybridization.- Proteomics of Brain Endothelium.- Immunoblot Detection of Brain Vascular Proteins.- Immunohistochemical Detection of Endothelial Proteins.- Genetically Altered Mice.- Cerebral Vascular Function in Genetically Altered Mice.- Generation of Transgenic Mice by Pronuclear Injection.- Mouse Genome Modification.
