Journal of Neurocytology

, Volume 25, Issue 1, pp 645–657 | Cite as

Ultrastructural study on the interaction of native and cationized albumin-gold complexes with mouse brain microvascular endothelium

  • A. W. Vorbrodt
  • D. H. Dobrogowska
  • A. S. Lossinsky
Article

Summary

The main objective of this ultrastructural study was to gain insights into the cellular mechanisms responsible for the enhanced brain uptake of blood-borne cationized albumin observed by several authors utilizing quantitative methodology. Mice were injected intravenously or into the common carotid artery (in vivo experiments) or perfusedin situ with solutions of native or cationized bovine serum albumin complexed with colloidal gold (BSA-G or cBSA-G respectively). The results indicate that: (1) the main drawbacks ofin vivo experiments are very intense phagocytosis of the tracer particles by Kupffer cells located in the liver sinusoids and also escape of the tracer through capillaries of skeletal and heart muscles. This results in a rapid decline of the concentration and disappearance of the circulating tracer particles; (2) BSA-G and cBSA-G bothin vivo (up to 30 min circulation) or after perfusionin situ (up to 15 min) do not cross the wall of brain microvessels representing the blood-brain barrier; (3) enhanced entrance of cationized albumin into the brain occurs through fenestrated endothelium of the capillaries located in the examined circumventricular organs (median eminence and neurohypophysis). Although BSA-G is also transported by these fenestrated capillaries, this process is evidently less intense than in the case of cBSA-G; (4) the enhanced passage of cBSA-G across fenestrated capillaries occurs mainly via vesicular transport (adsorptive transcytosis), through transendothelial channels and eventually through interendothelial junctional clefts; (5) the fenestrated capillaries of the choroid plexus appear to be less permeable for both tracers than those located in the other circumventricular organs.

Keywords

Colloidal Gold Tracer Particle Median Eminence Brain Uptake Vesicular Transport 

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References

  1. Balin, B. J. &Broadwell, R. D. (1988) Transcytosis of protein through the mammalian cerebral epithelium and endothelium. I. Choroid plexus and the blood-cerebrospinal fluid barrier.Journal of Neurocytology 17, 809–26.Google Scholar
  2. Bergmann, P., Kacenelenbogen, R. &Vizet, A. (1984) Plasma clearance, tissue distribution and catabolism of cationized albumins with increasing isoelectric points in the rat.Clinical Science 67, 35–43.Google Scholar
  3. Broadwell, R. D. &Sofroniew, M. V. (1993) Serum proteins bypass the blood-brain fluid barriers for extracellular entry to the central nervous system.Experimental Neurology 120, 245–63.Google Scholar
  4. Dermietzel, R., Thürauf, N. &Kalweit, P. (1983) Surface charges associated with fenestrated brain capillaries. II.In vivo studies on the role of molecular charge in endothelial permeability.Journal of Ultrastructure Research 84, 111–19.Google Scholar
  5. Frens, G. (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold suspension.Nature 241, 20–2.Google Scholar
  6. Ghitescu, L., Fixman, A., Simionescu, M. &Simionescu, N. (1986) Specific binding sites for albumin restricted to plasmalemmal vesicles of continuous capillary endothelium: receptor-mediated transcytosis.Journal of Cell Biology 102, 1304–11.Google Scholar
  7. Handley, D. A. &Chien, S. (1987) Colloidal gold labelling studies related to vascular and endothelial function, hemostasis and receptor-mediated processing of plasma macromolecules.European Journal of Cell Biology 43, 163–74.Google Scholar
  8. Horrisberger, M. &Rosset, J. (1977) Colloidal gold, a useful marker for transmission and scanning electron microscopy.Journal of Histochemistry and Cytochemistry 25, 295–305.Google Scholar
  9. Kumagai, A. K., Eisenberg, J. B. &Pardridge, W. M. (1987) Absorptive-mediated endocytosis of cationized albumin and a β-endorphin-cationized albumin chimeric peptide by isolated brain capillaries.Journal of Biological Chemistry 262, 15214–19.Google Scholar
  10. Milici, A. J., Watrous, N. E., Stukenbrok, H. &Palade, G. E. (1987) Transcytosis of albumin in capillary endothelium.Journal of Cell Biology 105, 2603–12.Google Scholar
  11. Mühlpfordt, H. (1982) The preparation of colloidal gold particles using tannic acid as an additional reducing agent.Experientia 38, 1127–8.Google Scholar
  12. Pardridge, W. M., Eisenberg, J. &Cefalu, W. T. (1985) Absence of albumin receptor on brain capillariesin vivo andin vitro.American Journal of Physiology 249, E264–7.Google Scholar
  13. Pardridge, W. M., Kumagai, A. K. &Eisenberg, J. B. (1987) Chimeric peptides as a vehicle for peptide pharmaceutical delivery through the blood-brain barrier.Biochemical and Biophysical Research Communications 146, 307–13.Google Scholar
  14. Pardridge, W. M., Triguero, D., Buciak, J. &Yang, J. (1990) Evaluation of cationized rat albumin as a potential blood-brain barrier drug transport vector.Journal of Pharmacology and Experimental Therapeutics 255, 893–9.Google Scholar
  15. Reese, T. S. &Karnovsky, M. J. (1967) Fine structural localization of a blood-brain barrier to exogeneous peroxidase.Journal of Cell Biology 34, 207–17.Google Scholar
  16. Schmidley, J. W. &Wissig, S. L. (1986) Anionic sites on the luminal surface of fenestrated and continuous capillaries.Brain Research 363, 265–71.Google Scholar
  17. Shen, W. C. &Ryser, H. J. (1978) Conjugation of poly-l-lysine to albumin and horseradish peroxidase: a novel method of enhancing the cellular uptake of proteins.Proceedings of National Academy of Sciences (USA) 75, 1872–6.Google Scholar
  18. Shimon-Hophy, M., Wadhwani, K. C., Chandrasekaran, K., Larson, D., Smith, Q. R. &Rapoport, S. I. (1991) Regional blood-brain barrier transport of cationized bovine serum albumin in awake rats.American Journal of Physiology 261, R478–83.Google Scholar
  19. Simionescu, N., Simionescu, M. &Palade, G. E. (1981) Differential microdomains on the luminal surface of the capillary endothelium. I. Preferential distribution of anionic sites.Journal of Cell Biology 90, 605–13.Google Scholar
  20. Simionescu, M., Ghitescu, L., Fixman, A. &Simionescu, N. (1987) How plasma macromolecules cross the endothelium.News in Physiological Sciences 2, 97–100.Google Scholar
  21. Smith, K. R. &Borchardt, R. T. (1989) Permeability and mechanism of albumin, cationized albumin, and glycosylated albumin transcellular transport across monolayers of cultured bovine brain capillary endothelial cells.Pharmaceutical Research 6, 466–73.Google Scholar
  22. Villaschi, S. (1989) Preparation and application of albumin-gold complex. InColloidal gold, Vol. 2, (edited byHayat, M. A.) pp. 163–74. New York: Academic Press.Google Scholar
  23. Vorbrodt, A. W., Lossinsky, A. S. (1986) Transport of homologous albumin through various capillary endothelia in mice.Journal of Cell Biology 103, 193a (Abstract).Google Scholar
  24. Vorbrodt, A. W. &Trowbridge, R. S. (1991) Ultrastructural study of transcellular transport of native and cationized albumin in cultured sheep brain microvascular endothelium.Journal of Neurocytology 20, 998–1006.Google Scholar
  25. Vorbrodt, A. W., Lossinsky, A. S. &Wisniewski, H. M. (1987) Ultrastructural studies of the uptake and transport of homologous albumin by brain endothelia.Canadian Journal of Neurological Sciences 14, 342 (Abstract).Google Scholar
  26. Vorbrodt, A. W., Lossinsky, A. S., Dobrogowska, D. H. &Wisniewski, H. M. (1993) Cellular mechanisms of the blood-brain barrier opening to albumin-gold complex.Histology and Histopathology 8, 51–61.Google Scholar
  27. Vorbrodt, A. W., Dobrogowska, D. H. &Lossinsky, A. S. (1994) Ultrastructural study on the interaction of insulin-albumin-gold complex with mouse brain microvascular endothelial cells.Journal of Neurocytology 23, 201–8.Google Scholar
  28. Vorbrodt, A. W., Dobrogowska, D. H., Ueno, M. &Lossinsky, A. S. (1995) Immunocytochemical studies of protamine-induced blood-brain barrier opening to endogenous albumin.Acta neuropathologica 89, 491–9.Google Scholar
  29. Zlokovic, B. V. (1990)In vivo approaches for studying peptide interactions at the blood-brain barrier.Journal of Controlled Release 13, 185–201.Google Scholar

Copyright information

© Chapman and Hall 1996

Authors and Affiliations

  • A. W. Vorbrodt
    • 1
  • D. H. Dobrogowska
    • 1
  • A. S. Lossinsky
    • 1
  1. 1.New York State Office of Mental Retardation and Developmental DisabilitiesInstitute for Basic Research in Developmental DisabilitiesStaten IslandUSA

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