Skip to main content
Log in

Diabetes-related changes in the protein composition of rat cerebral microvessels

  • Original Articles
  • Published:
Neurochemical Research Aims and scope Submit manuscript

Abstract

To determine the effect of diabetes mellitus on cerebral microvessel protein composition, post translational modification of proteins with glucose and malondialdehyde (MDA) was determined and the abundant protein species found in cerebral microvessels isolated from control and streptozotocin-induced diabetic rats were studied. Two dimensional gel electrophoresis and computer assisted densitometry revealed that only one out of 25 quantitated proteins was significantly altered in diabetic rats after 5 weeks of uncontrolled hyperglycemia. The level of glycosylation of cerebral microvessel protein mixture was significantly increased in diabetic rats compared to control rats (168.8±25 vs 109.5±4.8 nmol/mg) (p<0.05). Western blot analysis of cerebral microvessel proteins from diabetic rats using a specific antibody against MDA-modified proteins revealed three protein spots with molecular weights of approximately 60,000 Kd. These were shown not to be contaminants from cerebral tissue or plasma proteins modified with MDA. It is concluded that short duration of streptozotocin-induced diabetes mellitus in rats is associated with some qualitative changes in protein composition of cerebral microvessels. These changes may contribute to the diabetes-related alterations in the blood-brain barrier.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. McCall, A. 1992. The impact of diabetes on the central nervous system. Diabetes 41:557–570.

    Google Scholar 

  2. Mooradian, A. D. 1988. Diabetic complications of the central nervous system. Endocrine Rev. 9:346–356.

    Google Scholar 

  3. Mooradian, A. D., and Mariash, C. N. 1987. Effects of insulin and glucose on cultured rat hepatocyte gene expression. Diabetes 36:938–943.

    Google Scholar 

  4. Brownlee, M., Vlassera, H., and Cerami, A. 1984. Non enzymatic glycosylation and the pathogenesis of diabetic complications. Ann. Int. Med. 101:527–537.

    Google Scholar 

  5. Baynes, J. W. 1991 Role of oxidative stress in development of complications in diabetes. Diabetes 40:405–412.

    Google Scholar 

  6. Mooradian, A. D., Scarpace, P. J. 1992. Beta-adrenergic receptor activity of cerebral microvessels in experimental diabetes mellitus. Brain Res. 583:155–160.

    Google Scholar 

  7. Goldstein, G. W., Wolinsky, J. S., Csejtey, J., and Diamond, I. 1975. Isolation of metabolically active capillaries from rat brain. J. Neurochem. 25:715–717.

    Google Scholar 

  8. Orlowski, M., and Meister, A. 1965. Isolation of gamma glutamyl transpeptidase from hog kidney. J. Biol. Chem. 240:338–347.

    Google Scholar 

  9. Lowry, O. H., Rosebrough, J. N., Farr, A. L., and Randall, R. J. 1951. Protein measurement with the folin phenol reagent. J. Biol. Chem. 193:262–275.

    Google Scholar 

  10. Gallop, P. M., Fluckiger, R., Hannekan, A., Mininsohn, M. M., and Gabbay, K. H. 1981. Chemical quantitation of hemoglobin glycosylation. Fluorometric detection of formaldehyde released upon periodate oxidation of glycoprotein. Anal. Biochem. 117:427–432.

    Google Scholar 

  11. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685.

    Google Scholar 

  12. O'Farrell, P. H. 1975. High resolution two-dimensional electrophoresis of protein. J. Biol. Chem. 250:4007–4021.

    Google Scholar 

  13. Oakley, B. R., Kirisch, D. R., and Moris, N. R. 1980. A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal. Biochem. 105:361–363.

    Google Scholar 

  14. Mooradian, A. D., Meredith, K. E. 1992. The effect of age on protein composition of rat cerebral microvessels. Neurochem. Res. 17:665–670.

    Google Scholar 

  15. Towbin, H., Staehelin, T., and Gordon, J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets. Procedure and some applications. Proc. Natl. Acad. Sci. USA, 76:4350–4354.

    Google Scholar 

  16. Lung, C. C., Fleisher, J. H., Meinke, G. C., Pinnas, J. L. 1990. Immunochemical properties of malondialdehyde-protein adducts. J. Immunol. Methods 128:127–132.

    Google Scholar 

  17. Brada, D., and Roth, J. 1984. “Golden blot”. Detection of polyclonal and monoclonal antibodies bound to antigens or nitrocellulose by Protein A-gold complexes. Anal. Biochem. 142:79–83.

    Google Scholar 

  18. Surek, B. and Latzko, E. 1984. Visualization of antigenic proteins blotted onto nitrocellulose using the immuno-gold-staining (IGS)-method. Biochem. Biophys. Res. Commun. 121:284–289.

    Google Scholar 

  19. Sell, D. R., and Monnier, V. M. 1989. Structure elucidation of a senescent crosslink from human extracellular matrix: Implications of ribose in the aging process. J. Biol. Chem. 264:21597–21602.

    Google Scholar 

  20. Mooradian, A. D., and Smith, T. L. 1992. The effect of experimentally-induced diabetes mellitus on the lipid order and composition of rat cerebral microvessels. Neurosci. Lett. 145:145–148.

    Google Scholar 

  21. Lung, C. C., Pinnas, J. L., Yahya, M. D., Meinke, G. C., Mooradian, A. D. 1993. Malondialdehyde modified proteins and their antibodies in the plasma of control and streptozotocin induced diabetic rats. Life Sci. 52:329–337.

    Google Scholar 

  22. Tayarani, I., Chaudiere, J., Lefauconnier, J. M., Bourre, J-M. 1987. Enzymatic protection against peroxidative damage in isolated brain capillaries. J. Neurochem. 48:1399–1402.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mooradian, A.D., Pinnas, J.L., Lung, CC. et al. Diabetes-related changes in the protein composition of rat cerebral microvessels. Neurochem Res 19, 123–128 (1994). https://doi.org/10.1007/BF00966805

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00966805

Key Words

Navigation