Opening of the blood-brain barrier in Anolis carolinensis. A high voltage electron microscope protein tracer study

Abstract

The tight junctions between endothelial cells of capillaries in the forebrain of Anolis carolinensis are a common component of the structural basis for the blood-brain barrier in this reptile. The complexity of these junctions, which is apparent in platinum replicas of freeze-fractured brain capillaries, is unchanged by treatments designed to render the blood-brain barrier of these lizards leaky to horseradish peroxidase. An alternative route for extravasation of horseradish peroxidase, following injection of chameleons with 2.7 mg of D-glucose to render their brain capillaries leaky, is a system of transient cytoplasmic vesicles and vesiculo-tubular channels whose lumina may be open to the luminal or abluminal surface (or both) of the capillary endothelial cell. High voltage electron microscopy (HVEM) of 0.25 and 0.5 micron thick plastic sections of experimental brain capillary endothelium confirmed the existence of vesiculo-tubular conduits. These channels display a sigmoid morphology and are situated in the cytoplasm at angles oblique to the luminal and abluminal surfaces of the endothelium. Occasionally, the channels spanned the entire endothelial wall of the capillary, and in such cases, appeared to connect the lumen with the brain extracellular compartment. HVEM images (including stereo pairs) of the vesiculo-tubular channels show them to have a scalloped, irregular profile consistent with their proposed formation by fusion of pinocytotic vesicles. Also, HVEM examinations of experimental capillaries from peroxidase-treated lizards reveal massive quantities of dense reaction product in cytoplasmic vesicles and vesiculo-tubular membrane compartments of the endothelium, and the complex pleomorphism exhibited by these structures. Observations made in the present study suggest that as a consequence of severe hyperglycaemia, transendothelial channels form in the brain capillaries by fusion of pinocytotic vesicles generated by accelerated pinocytosis at the luminal surface of the endothelium, and subsequently serve as open routes for massive floods of tracer into the central nervous system.