Chapter 9. Subcellular Fractionation Studies on the Cells Isolated from Normal and Atherosclerotic Aorta

  1. Ruth Porter and
  2. Julie Knight
  1. T. J. Peters,
  2. T. Takano and
  3. C. De Duve

Published Online: 30 MAY 2008

DOI: 10.1002/9780470719954.ch9

Ciba Foundation Symposium 12 - Atherogenesis: Initiating Factors

Ciba Foundation Symposium 12 - Atherogenesis: Initiating Factors

How to Cite

Peters, T. J., Takano, T. and De Duve, C. (2008) Subcellular Fractionation Studies on the Cells Isolated from Normal and Atherosclerotic Aorta, in Ciba Foundation Symposium 12 - Atherogenesis: Initiating Factors (eds R. Porter and J. Knight), John Wiley & Sons, Ltd, Chichester, UK. doi: 10.1002/9780470719954.ch9

Author Information

  1. The Rockefeller University, New York

Publication History

  1. Published Online: 30 MAY 2008
  2. Published Print: 1 JAN 1973

ISBN Information

Print ISBN: 9789021940137

Online ISBN: 9780470719954

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Keywords:

  • subcellular fractionation;
  • atherosclerotic aorta;
  • triglyceride lipase;
  • acid phosphatase;
  • atheromatous tissue

Summary

Using the cholesterol-fed rabbit as a model of atherosclerosis we have investigated some of the subcellular events that accompany atherogenesis. A method was developed for the isolation and one-step subcellular fractionation of smooth muscle cells, from normal and atheromatous aorta. Compared to control cells, atheromatous cells showed a striking increase in certain lysosomal enzymes (β-galactosidase, β-glucuronidase and cathepsin D) which parallelled the marked increase in their cholesterol content. Other lysosomal enzymes showed a smaller increase (N-acetyl-β-hexosaminidases, acid phosphatase, cholesterol esterase and triglyceride lipase).

Subcellular fractionation of cells from atheromatous tissue showed the presence of a second population of low-density lysosomes with a distinct enzyme content. Compared to lysosomes isolated from normal arteries, these low-density particles contained little acid phosphatase and cholesterol esterase but were relatively rich in N-acetyl-β-glucosaminidase, cathepsin D, β-galactosidase and triglyceride lipase. They also contained considerable amounts of cholesterol and cholesterol ester. In contrast, there were no changes in the specific activity or behaviour in sucrose density gradients of mitochondrial or plasma membrane marker enzymes.

The application of this isolation and fractionation technique to a sample of human aorta is described and the possible role of the lysosome in the atherosclerotic process is discussed.