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Cholesterol distribution and structural differentiation in the sarcoplasmic reticulum of rat cardiac muscle cells

A freeze-fracture cytochemical investigation

Summary

The polyene compound, filipin, was used as a probe to localize cholesterol in the membranes of the rat cardiac muscle cell, with particular reference to the sarcoplasmic reticulum (SR). Filipin binds specifically to cholesterol (and related 3-β-hydroxysterols) in membranes, producing distinct deformations which can be viewed by freeze-fracture and used as markers for the presence of cholesterol-rich regions in the membrane plane. In freeze-fracture replicas of filipin-treated rat myocardium, the muscle cells revealed abundant deformations in their plasma membranes, no deformations in mitochondrial membranes, and an intermediate response in the SR. These results are in agreement with the levels of cholesterol reported in isolated fractions of the different membrane types, and confirm the specificity of filipin action. Within the SR, the filipin-induced deformations were not randomly distributed but occurred more commonly in free SR at or near the Z-region of the sarcomere than in other parts of the free SR or the junctional SR. This finding is interpreted as evidence for a non-homogeneous distribution of cholesterol in cardiac muscle cell SR. The possible significance of cholesterol in relation to structural differentiation and function of the SR is discussed.

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Correspondence to .Dr. N. J. Severs.

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Severs, N.J. Cholesterol distribution and structural differentiation in the sarcoplasmic reticulum of rat cardiac muscle cells. Cell Tissue Res. 224, 613–624 (1982). https://doi.org/10.1007/BF00213756

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Key words

  • Sarcoplasmic reticulum
  • Cardiac muscle cells (rat)
  • Cholesterol
  • Filipin
  • Freeze-fracture cytochemistry