Summary
Serum proteins as constituents of vasogenic brain edema were visualized both macroscopically and microscopically applying a double layer immunofluorescence technique to paraffin embedded material derived from three experimental series: peritumorous edema following xeno-transplantation of glioma cells, edema after cerebral embolization with micropheres, and edema after unilateral MCA occlusion. Exclusively cats were used as experimental animals.
The staining procedures resulted in selective green fluorescence of vessel contents as well as edema protein, which was demonstrable even macroscopically at times, where edema formation reaches a maximum in each experimental series. Microscopically, serum protein could be traced up to the end of observation time ranging from 1 to 4 weeks, where the specific fluorescence was related to cellular structures.
As compared to other techniques employed in brain edema localization, immunostaining mainly offers the following advantages: avoidance of in vivo tracing, better structural resolution in paraffin than in freeze sections, high specificity and sensitivity in antigen localization.
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Allen, T. H., orahovats, P. D.: Combination of toluidine dyes with plasma albumin. Am. J. Physiol.161, 473–482 (1950)
Baker, R. N., Cancilla, P. A., Pollock, P. S., Frommes, B. S.: The movement of exogenous protein in experimental cerebral edema. J. Neuropathol. Exp. Neurol30, 668–679 (1971)
Bignami, A., Eng, L. F., Uyeda, C. T.: Localization of the glial fibrillary protein in astrocytes by immunofluorescence. Brain Res.43, 429–435 (1972)
Blakemore, W. F.: The fate of escaped plasma protein after thermal necrosis of the rat brain: an electron microscope study. J. Neuropathol. Exp. Neurol.28, 139–152 (1969)
Blinderman, E. E., Markham, Ch. H.: Fluoresein studies of cerebral edema producted by the cryogenic probe. J. Neurosurg.23, 38–44 (1965)
Böhme, G.: Untersuchungen an der area postrema von Gallus domesticus. Acta Neuropathol. (Berl.)21, 308–315 (1972)
Bolton, A. E., Hunter, W. M.: The labelling of proteins to high specific radioactivities by conjugation to a125I-containing alcylating agent. J. Biochem.133, 529–539 (1973)
Brightman, M. W., Klatzo, I., Olsson, Y., Reese, T. S.: The bloodbrain barrier to protein under normal and pathological conditions. J. Neurol. Sci.10, 215–239 (1970)
Broman, T., Radner, S., Svanberg, L.: The duration of experimental disturbances in the cerebrovascular permeability due to circumscribed gross damage of the brain. Acta Psychiat. Neurol.24, 167–173 (1949)
Broman, T., Edström, R., Steinwall, O.: Technical aspects on dyes and radiotracers in the determination of blood-brain barrier damage. Acta Psych. Neurol. Scand.36, 69–75 (1961)
Broman, T., Branemark, P.-I., Johansson, B., Steinwall, O.: Intravital and postmortem studies on air embolism damage of the blood-brain barrier tested with trypan blue. Acta. Neurol. Scand.42, 146–152 (1956)
Bryar, G. E., Goldstein, N. P., Svien, H. J., Sayre, G. P., Jones, J. D.: Experimental cerebral edema: Vital staining with Evans blue during the development and regressive phase. J. Neurosurg.30, 391–398 (1969)
Clasen, R. A., Cooke, P. M., Pandolfi, S., Boyd, D., Raimondi, A. J.: Experimental cerebral edema produced by focal freezing. J. Neuropathol. Exp. Neurol.21, 579–596 (1962)
Clasen, R. A., Cooke, P. M., Pandolf, S., Boyd, D., Raimondi, A. J.: Experimental cerebral edema produced by focal freezing. J. Neuropathol. Exp. Neurol.21, 579–596 (1962)
Clasen, R. A., Cooke, P. M., Pandolf, S., Boyd, D., Raimondi, A. J.: Vital staining mechanisms in the damged and edematous central nervous system. Meeting abstract. 38th Annual Meeting of the American Association of Neuropathologists, Atlantic City, N.J., June 16, 1962. J. Neuropathol. Exp. Neurol.22, 318–363 (1963)
Clasen, R. A., Pandolfi, S., Hass, G. M.: Vital staining serum albumin and the blood-brain barrier. J. Neuropathol. Exp. Neurol.29, 266–284 (1970)
Coons, A. H., Kaplan, M. H.: Localization of antigen in tissue cells. J. Exp. Med.91, 1–13 (1950)
Cutler, R. W., Watters, G. V., Barlow, Ch. W.:125I-labeled protein in experimental brain edema. Arch. Neurol.11, 225–238 (1964)
Edvinsson, L., Nielson, K. C., Owman, Ch., West, K. A.: Alterations in intracranial pressure, blood-brain barrier, and brain edema after sub-chronic implantation of a cannula into the brain of conscious animals. Acta Physiol. Scand.82, 527–531 (1971)
Ehrlich, P.: Das Sauerstoffbedürfnis des Organismus. In: Eine farbenanalytische Studie. Berlin 1885
Esiri, M. M., Taylor, C. R., Mason, D. Y.: Application of an immunoperoxydase method to a study of the central nervous system: preliminary findings in a study of human formalin-fixed material. Neuropath. Appl. Neurobiol.2, 233–246 (1976)
Gregersen, M. I., Rawson, R. A.: The disappearance of T-1824 and structurally related dyes from the blood stream. Am. J. Physiol.138, 698–707 (1943)
Hamberger, A., Hamberger, B.: Uptake of catecholamines and penetration of trypan blue after blood-brain barrier lesions. Z. Zellforschung70, 386–392 (1966)
Hirano, A., Dembitzer, H. M., Becker, N. H., Levine, S., Zimmermann, H. M.: Fine structural alterations of the bloodbrain barrier in experimental allergic encephalomyelitis. J. Neuropathol. Exp. Neurol.29, 432–440 (1970)
Hossmann, K.-A., Olsson, Y.: The effect of transient cerebral ischemia on the vascular permeability to protein tracers. Acta Neuropathol. (Berl.)18, 103–112 (1971)
Hunter, W. M., Greenwood, F. C.: Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature194, 495–496 (1962)
Klatzo, I., Miquel, J.: Observation on pinocytosis in nervous tissue. J. Neuropathol. Exp. Neurol.19, 475–487 (1960)
Klatzo, I.: Presidential address. Neuropathological aspects of brain edema. J. Neuropathol. Exp. Neurol.26, 1–14 (1967)
Klatzo, I.: Application of fluorescence microscopy to the study of the nervous system. In: The central nervous system. International Academy of Pathology Monograph No. 9, p. 319. Baltimore: Williams and Wilkins 1968
Klatzo, I., Piraux, A., Laskowski, E. J.: The relationship between tissue elements, blood-brain barrier and edema in a local brain injury. J. Neuropathol. Exp. Neurol.17, 548–564 (1958)
Klatzo, I., Miquel, J., Otenasek, R.: The application of fluorescein labelled serum protein to the study of vascular permeability in the brain. Acta Neuropathol. (Berl.)2, 144–160 (1962)
Lee, J. C., Olszewski, J.: Effect of air embolism on permeability of cerebral blood vessels. Neurology (Minneap.)9, 614–625 (1959)
Ludwin, S. K., Kosek, J. C., Eng, L. F.: The topographical distribution of S-100 and GFA proteins in the adult rat brain: an immunohistochemical study using horseradish peroxydase-labelled antibodies. J. Comp. Neurol.165, 197–208 (1976)
Lumsden, C. E.: The immunogenesis of the multiple sclerosis plaque. Brain Res.28, 365–390 (1971)
Moriarty, G. C., Moriarty, M. C., Sternberger, L. A.: Ultrastructural immunocytochemistry with unlabelled antibodies and the peroxydase-antiperoxidase complex. J. Histochem. Cytochem.21, 825–833 (1973)
Oldstone, M. B., Dixon, F. J.: Immunohistochemical study of allergic encephalomyelitis. Am. J. Pathol.52, 251–263 (1968)
Olsson, Y., Crowell, R. M., Klatzo, I.: The blood-brain barrier to protein tracers in focal cerebral ischemia and infarction caused by occlusion of the middle cerebral artery. Acta Neuropathol. (Berl.)18, 89–102 (1971)
Rapoport, S. I.: Reversible opening of the blood-brain barrier. Science173, 1026–1028 (1971)
Rawson, R. A.: The binding of T-1824 and structurally related diazo dyes by the plasma proteins. Am. J. Physiol.138, 708–717 (1943)
Reese, T. S., Karnowsky, M. J.: Fine structural localization of a blood-brain to exogenous peroxydase. J. Cell Biol.34, 207–217 (1967)
Rinder, L., Olsson, Y.: Studies on permeability changes in experimental brain concussion. Acta Neuropathol. (Berl.)11, 201–209 (1968)
Tavolato, B. F.: Immunoglobulin G distribution in multiple sclerosis brain. J. Neurol. Sci.24, 1–11 (1975)
Tschirgi, R. D.: Protein complexes and the impermeability of the blood-brain barrier to dyes. Am. J. Physiol.163, 756 (1950)
Westergaard, E., Brightman, M. W.: Transport of proteins across normal cerebral arterioles. J. Comp. Neurol.152, 17–44 (1973)
Woyciechowska, J. L., Brzosko, W. J.: Immunofluorescence study of brain plaques from two patients with multiple sclerosis. Neurology (Minneap.)27, 620–622 (1977)
Vise, W. M., Liss, L., Yashon, D., Hunt, W. E.: Astrocytic processes: a route between vessels and neurons following blood-brain barrier injury. J. Neuropathol. Exp. Neurol.24, 324–334 (1975)
Vulpe, M., Hawkins, A., Rozdilsky, B.: Permeability of cerebral blood vessels in experimental allergic encephalomyelitis studied by radioactive iodinated bovine albumin. Neurology (Minneap.)10, 171–177 (1960)
Weller, T. H., Coons, A. H.: Fluorescent antibody studies with agents of varicella and herpes zoster-propagated in vitro. Proc. Soc. Exp. Biol. Med.86, 789–794 (1954)
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Wilmes, F., Hossmann, K.A. A specific immunofluorescence technique for the demonstration of vasogenic brain edema in paraffin embedded material. Acta Neuropathol 45, 47–51 (1979). https://doi.org/10.1007/BF00691804
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DOI: https://doi.org/10.1007/BF00691804