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Intramembrane particles and filipin labelling on the membranes of autophagic vacuoles and lysosomes in mouse liver

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Summary

Morphologically detectable protein (intramembrane particles) and cholesterol (filipin labelling) in the membranes of autophagic vacuoles and lysosomes were studied in mouse hepatocytes using thin-section and freeze-fracture electron microscopy. Both isolated autophagic vacuoles and lysosomes, and intact tissue blocks were used due to the facts (i) that lysosomes are difficult to recognize in freeze-fracture replicas of intact hepatocytes, and (i) that filipin penetration into the tissue blocks is unsatisfactory. Intramembrane particle density was low in the membranes of early autophagic vacuoles (defined as round-shaped vacuoles in which an inner membrane parallel with the outer limiting membrane was clearly visible). The lysosomal membranes contained considerably more intramembrane particles. Particle-rich lysosomes or other vesicles were observed to fuse with the early autophagic vacuoles. The membranes of nascent autophagic vacuoles with morphologically intact contents were usually not labelled by filipin, whereas the membranes of all other autophagic vacuoles and lysosomes were heavily labelled. The increased cholesterol in the membranes of slightly older autophagic vacuoles is presumably derived from cholesterol-rich lysosomes or other vesicles fusing with the vacuoles and from the degrading organelles inside the autophagic vacuoles.

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Authors and Affiliations

  1. Department of Cell Biology, University of Jyväskylä, Jyväskylä, Finland

    Eeva-Liisa Punnonen & Kari Mattila

  2. Laboratory of Electron Microscopy, Department of Biology, University of Turku, Turku, Finland

    Kaarina Pihakaski

  3. Department of Electron Microscopy, University of Helsinki, Helsinki, Finland

    Kari Lounatmaa

  4. Farmos Group Ltd, Research Center, Turku, Finland

    Pirkko Hirsimäki

Authors
  1. Eeva-Liisa Punnonen
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  2. Kaarina Pihakaski
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  3. Kari Mattila
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  4. Kari Lounatmaa
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  5. Pirkko Hirsimäki
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Punnonen, EL., Pihakaski, K., Mattila, K. et al. Intramembrane particles and filipin labelling on the membranes of autophagic vacuoles and lysosomes in mouse liver. Cell Tissue Res. 258, 269–276 (1989). https://doi.org/10.1007/BF00239447

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