Sulfatide and sphingomyelin loading of living cells as tools for the study of ceramide turnover by lysosomal ceramidase--implications for the diagnosis of Farber disease.

The ceramide turnover by lysosomal ceramidase in intact, living cells was investigated by loading radiolabeled sulfatide or sphingomyelin in situ on skin fibroblasts and lymphoid cells. The cells originated from normal individuals and from patients with acid ceramidase deficiency (Farber disease). While fibroblasts from individuals with Farber disease exhibited some impairment in the degradation of the ceramide produced by sulfatide hydrolysis, lymphoid cells from individuals with Farber disease metabolized the ceramide as readily as did normal cells, suggesting the existence in lymphoid cells of a non-lysosomal degradation pathway for the sulfatide-derived ceramide. In contrast, sphingomyelin loading in the presence of serum showed a considerably decreased turnover of ceramide in both fibroblasts and lymphoid cells from individuals with Farber disease. Further methodologic variation led to the use of LDL-associated radioactive sphingomyelin; LDL-association promoted the targeting of exogenous sphingomyelin to lysosomes. As a result, an almost complete deficiency of ceramide degradation was found in cells from severely affected patients with Farber disease. Our data with this novel method show that sphingomyelin loading of intact living cells is a simple, alternative means for determining ceramide degradation by lysosomal ceramidase and for diagnosing Farber disease.
AuthorsT Levade, M C Tempesta, H W Moser, A H Fensom, K Harzer, A B Moser, R Salvayre
JournalBiochemical and molecular medicine (Biochem Mol Med) Vol. 54 Issue 2 Pg. 117-25 (Apr 1995) ISSN: 1077-3150 [Print] UNITED STATES
PMID8581356 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Ceramides
  • Sphingomyelins
  • Sulfoglycosphingolipids
  • Tritium
  • Amidohydrolases
  • ASAH1 protein, human
  • Acid Ceramidase
  • Ceramidases
  • Acid Ceramidase
  • Amidohydrolases (deficiency, metabolism)
  • Cell Transformation, Viral
  • Cells, Cultured
  • Ceramidases
  • Ceramides (metabolism)
  • Evaluation Studies as Topic
  • Female
  • Fibroblasts (cytology, metabolism)
  • Herpesvirus 4, Human
  • Humans
  • Lipid Metabolism
  • Lymphocyte Activation
  • Lysosomal Storage Diseases (diagnosis, drug therapy, metabolism)
  • Lysosomes (enzymology, metabolism)
  • Male
  • Sphingomyelins (chemistry, metabolism, pharmacology)
  • Sulfoglycosphingolipids (chemistry, metabolism, pharmacology)
  • Time Factors
  • Tritium

Join CureHunter, for free Research Interface BASIC access!

Take advantage of free CureHunter research engine access to explore the best drug and treatment options for any disease. Find out why thousands of doctors, pharma researchers and patient activists around the world use CureHunter every day.
Realize the full power of the drug-disease research network!

Choose Username:
Verify Password: