The Histochemical Journal

, Volume 16, Issue 9, pp 941–954 | Cite as

Immunocytochemistry of lysosomal hydrolases and their precursor forms in normal and mutant human cells

  • J. M. Van Dongen
  • R. A. Barneveld
  • H. J. Geuze
  • H. Galjaard
Papers
Received:

Summary

The acid hydrolases α-glucosidase, β-galactosidase,N-acetyl-β-d-hexosaminidase, β-glucocerebrosidase and cathepsin D were studied immunocytochemically in normal and mutant human cells using monoclonal and affinity-purified polyclonal antibodies. For light microscopy, Rhodamine or Fluorescein-labelled conjugates were used, and for electron microscopy protein A-gold conjugates were employed. With the double labelling procedure, it was found that in normal fibroblasts every lysosome contained all the enzymes studied. The method described also enabled us to demonstrate the presence or absence of mutant enzyme protein in fibroblasts derived from patients with a genetic lysosomal enzyme deficiency.

Immunoreactive acid hydrolases or their precursor forms were found in the rough endoplasmic reticulum, the cisternae of the Golgi complex, Golgi associated vesicles and lysosomes. This is in agreement with the present concept that the Golgi complex plays an essential role in the processing and targeting of lysosomal enzymes.

Keywords

Rhodamine Lysosomal Enzyme Rough Endoplasmic Reticulum Golgi Complex Double Labelling 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Copyright information

© Chapman and Hall Ltd. 1984

Authors and Affiliations

  • J. M. Van Dongen
    • 1
  • R. A. Barneveld
    • 1
  • H. J. Geuze
    • 2
  • H. Galjaard
    • 1
  1. 1.Department of Cell Biology and GeneticsErasmus UniversityRotterdamThe Netherlands
  2. 2.Centre of Electron Microscopy, School of MedicineUniversity of UtrechtUtrechtThe Netherlands

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