Studies of Cerebral Capillary Endothelial Membrane

Drewes, L. D.; Lidinsky, William A.

doi:10.1007/978-1-4684-3752-2_2

Cited by 9 publications

(4 citation statements)

References 10 publications

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“…It was previously reported that the protein at band 3 incorporated significant levels of radioactivity during lumen labeling experiments (Drewes and Lidinsky, 1980). In these earlier experiments a phosphate-buffered saline perfusate was used.…”

Section: Discussionmentioning

confidence: 99%

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Characterization of the Blood‐Brain Barrier: Protein Composition of the Capillary Endothelial Cell Membrane

Lidinsky

Drewes

1983

Journal of Neurochemistry

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Microvessels were isolated from canine cerebral cortex, and the composition of the endothelial cell membrane was investigated. Endothelial cell membranes were separated from the surrounding basement membrane, solubilized, and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis in 12% gels. Staining with Coomassie Blue revealed a characteristic banding pattern of at least 12 major proteins with apparent molecular weights between 14,000 and 250,000. When proteins from red blood cell ghosts were run simultaneously, no similarities were observed, except for proteins at apparent molecular weights of 43,000 (band 3) and 35,000 (band 4). These two proteins migrated exactly to the positions of the erythrocyte proteins actin and glyceraldehyde 3-phosphate dehydrogenase, respectively. Membrane glycoproteins in gels were also examined by the use of fluorescent lectins. Of the fluorescein isothiocyanate-conjugated (FITC) lectins tested, only FITC-concanavalin A had an affinity for any membrane components. Diazotized [125I]iodosulfanilic acid, a membrane-impermeable reagent, was used to label the internal (lumen) cell surface and the external (antilumen) cell surface. Autoradiography and determination of radioactivity levels in gel slices showed that several proteins were specifically labeled, and that major differences in radioactivity of proteins existed in internal and external labeling experiments. It is concluded that the protein composition of the luminal membrane is different from that of the antiluminal membrane.

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Section: Discussionmentioning

confidence: 99%

“…T h e use of a membrane-impermeable labeling technique h a s allowed the differential labeling of t h e luminal a n d abluminal membranes. Some results have been presented in a n earlier communication (Drewes and Lidinsky, 1980).…”

Section: W a Lidinsky A N D L R Drewesmentioning

confidence: 98%

Characterization of the Blood‐Brain Barrier: Protein Composition of the Capillary Endothelial Cell Membrane

Lidinsky

Drewes

1983

Journal of Neurochemistry

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“…Over-homogenization results in extensive fragmentation of the MV, yielding structurally damaged MV. We found that sieving, when not preceded by a centrifugation gradient step [4,6,[16][17][18], allows contaminating ceils to remain in the final preparation. Both centrifugation and sieving (with the appropriate mesh size) are especially critical in the isolation of high quality MV from brain tumor tissue.…”

Section: Discussionmentioning

confidence: 93%

Isolation and characterization of microvessels from normal brain and brain tumors

Silbergeld

Ali‐Osman

1991

J Neuro-Oncol

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We describe a new technique for isolating microvessels from both brain and brain tumors. This method is relatively quick and provides a microvessel preparation free of contamination by other brain tissue. Using this method, structurally intact microvessels from normal rat brain and from a malignant rat astrocytoma were isolated and characterized with light microscopy, scanning electron microscopy and transmission electron microscopy. In contrast to microvessels derived from normal rat brain, rat astrocytoma microvessels had endothelial cells with multilayered basement membranes, extensive microvilli on the cell surfaces, and a significant increase in the number of pinocytes in the cytoplasm. Furthermore, astrocytoma microvessel endothelial cells had pleomorphic electron dense nuclei with pale perichromatin, whereas the nuclei of endothelial cells of microvessels derived from normal brain tissue were finely granular and homogeneous with characteristically electron dense perichromatin. The morphologic characteristics of the astrocytoma microvessels are similar to the histologic changes seen in astrocytoma tissue in situ, and correlate well with the known altered functions of brain tumor neovasculature.

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“…A procedure was adapted from previous techniques to isolate microvessels from rabbit brain under sterile conditions [1,3,32]. Capillaries were also obtained from mouse, rat, and guinea-pig brains using the same procedure.…”

Section: Capillariesmentioning

confidence: 99%

Interactions of tumor cells with intact capillaries: a model for intravasation

Repesh

Fitzgerald

1984

Clin Exp Metast

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The interactions of metastatic variants of the B16 and K-1735 melanoma cell lines with intact capillaries isolated from rabbit brain were studied in vitro. The abilities of various cells to attach to and flatten out on the surface of blood vessels were monitored with phase contrast and scanning electron microscopy. In general, cells highly metastatic in vivo were capable of attaching to and flattening out on the surface of capillaries at a faster rate than cells of low metastatic potential or normal cells. In addition, K-1735 melanoma cells and normal fibroblasts were labeled with 125I-iododeoxyuridine, incubated with capillaries, and subsequently passed through filters. This procedure separated unattached cells from cells attached to capillaries. This assay provided quantitative information on the adhesion of metastatic tumor cells and normal cells to capillaries. Tumor cells attached to capillaries in significantly greater numbers than normal cells. Cells with high metastatic potential attached to the capillaries in greater numbers than cells with low metastatic potential, but not significantly. This model may be useful for elucidating some of the mechanisms involved in tumor cell attachment and penetration of the capillary wall during intravasation in vivo.

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Studies of Cerebral Capillary Endothelial Membrane

Cited by 9 publications

References 10 publications

Characterization of the Blood‐Brain Barrier: Protein Composition of the Capillary Endothelial Cell Membrane

Characterization of the Blood‐Brain Barrier: Protein Composition of the Capillary Endothelial Cell Membrane

Isolation and characterization of microvessels from normal brain and brain tumors

Interactions of tumor cells with intact capillaries: a model for intravasation

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