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Ca2+ depletion-induced disconnection of tight junctions in isolated rat brain microvessels
Authors:Z. Nagy  U. G. Goehlert  L. S. Wolfe  I. Hüttner
Affiliation:(1) Dept. of Psychiatry, Semmelweis Medical University, Budapest, Hungary;(2) Montreal Neurological Institute, McGill University, Montreal, Canada;(3) Dept. of Pathology, McGill University, Montreal, Canada;(4) National Institutes of Health (NINCDS), Bldg. 36, Rm. 3B26, 20205 Bethesda, MD, USA
Abstract:Summary Cerebral microvessels were isolated from rat brains. One part of the microvessel pellets was incubated for 25 or 90 min in Krebs-Henseleit bicarbonate buffer (KHB) at pH 7.5 (control group). The other part of the pellets was treated for the same periods of time with Ca2+-free KHB, containing 2.2 mM EGTA and 2 mM glucose (experimental group). Morphological changes of endothelial tight junctions were evaluated in 100 randomly selected interendothelial clefts from isolated cerebral microvessels of each groups by electron microscopy. Following 25 min of incubation time, either with Ca2+-containing or with Ca2+-free KHB, no significant changes of tight junctions were observed. After 90 min of incubation in Ca2+-free medium, 58% of tight junctions were altered (in 42% partial, and in 16% complete disconnection of tight junctions were found). This contrasted the control group, where only 14% of tight junctions were disconnected (12% partially and 2% completely). Our results are consistent with a role for intercellular Ca2+ in maintaining structural integrity of cerebral tight junctions.Supported by the Medical Research Council of Canada, grant MT-5958 and the Hungarian Ministry of Health, grant 06/1-44/313
Keywords:Isolated cerebral capillaries  Ca2+ depletion  Ultrastructure of tight junctions
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