Distribution of lysosomal cathepsin D in normal, ischemic, and starved rabbit cardiac myocytes

R. S. Decker, A. R. Poole, K. Wildenthal

Research output: Contribution to journalArticle

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Abstract

Myocardial ischemia alters the immunofluorescent staining pattern of lysosomal cathepsin D from one that is essentially particulate to one which a diffuse staining pattern predominates. Prolonged starvation causes an apparently similar redistribution of cathepsin D from discrete secondary lysosomes to diffusely staining sites. However, because of the limits of resolution of immunofluorescence techniques, it has not been possible previously to define the exact nature and location of cathepsin D staining in these conditions. Accordingly, a new procedure employing peroxidase-labeled antibodies has been developed to allow localization of cathepsin D with electron microscopy. Most of the cathepsin D that can be detected in normal hearts with this technique is presented in classic lysosomal structures, analogous to the particulate staining pattern seen with immunofluorescence microscopy. The technique also reveals, for the first time, definitive ultrastructural evidence that ischemia induces lysosomal leakage and redistribution of cathepsin D into the cytoplasm. Starvation is not accompanied by release of cathepsin D into the cytoplasm; rather, there is widespread, increased distribution of cathepsin D within elements of the sarcoplasmic reticulum, whose diameters are too small for resolution with light microscopy. Thus, although both ischemia and starvation increase nonsedimentable cathepsin D activity, as measured biochemically, and produce a diffuse staining pattern under immunfluoresence microscopy, electron microscopy reveals a distinctly different subcellular distribution for this enzyme in the two conditions. These results emphasize that considerable caution must be exercised in interpreting biochemical and histochemical changes suggestive of enzyme release from lysosomes, in the absence of supportive ultrastructural evidence.

Original languageEnglish (US)
Pages (from-to)485-494
Number of pages10
JournalCirculation Research
Volume46
Issue number4
StatePublished - 1980

Fingerprint

Cathepsin D
Cardiac Myocytes
Rabbits
Staining and Labeling
Starvation
Lysosomes
Microscopy
Electron Microscopy
Cytoplasm
Ischemia
Sarcoplasmic Reticulum
Enzymes
Fluorescence Microscopy
Peroxidase
Myocardial Ischemia
Fluorescent Antibody Technique
Light
Antibodies

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Distribution of lysosomal cathepsin D in normal, ischemic, and starved rabbit cardiac myocytes. / Decker, R. S.; Poole, A. R.; Wildenthal, K.

In: Circulation Research, Vol. 46, No. 4, 1980, p. 485-494.

Research output: Contribution to journalArticle

Decker, RS, Poole, AR & Wildenthal, K 1980, 'Distribution of lysosomal cathepsin D in normal, ischemic, and starved rabbit cardiac myocytes', Circulation Research, vol. 46, no. 4, pp. 485-494.
Decker, R. S. ; Poole, A. R. ; Wildenthal, K. / Distribution of lysosomal cathepsin D in normal, ischemic, and starved rabbit cardiac myocytes. In: Circulation Research. 1980 ; Vol. 46, No. 4. pp. 485-494.
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