title: Phenotypic heterogeneity in cultured skin fibroblasts from patients with disorders of peroxisome biogenesis belonging to the same complementation group.
abstract: We have studied fibroblast cell lines derived from a control subject (cell line 85AD5035F) and three patients clinically described as having the Zellweger syndrome (cell line W78/515), the infantile form of Refsum disease (cell line BOV84AD) and hyperpipecolic acidaemia (cell line GM3605), respectively. The mutant cell lines belonged to the same complementation group. The fibroblasts were cultured under identical conditions and were harvested at different time intervals after reaching confluence. Several peroxisomal parameters were determined. In agreement with previous reports, a lowered enzymic activity of acyl-CoA: dihydroxyacetonephosphate acyltransferase and a decrease in latent catalase clearly distinguished the patient cell lines from the control cell line. However, the cell lines exhibited a phenotypic heterogeneity. This was most strikingly encountered when cells were processed for indirect immunofluorescence microscopy and stained with anti-(catalase). The control cells exhibited a punctate fluorescence, which is indicative of the presence of catalase in peroxisomes. In the mutant cell line W78/515 a diffuse fluorescence was observed, indicative of the presence of catalase in the cytosol. In the other two mutant cell lines a punctate fluorescence was observed in some of the cells. Moreover, clear differences in the extent of proteolytic processing of acyl-CoA oxidase were detected. The mutant cell line BOV84AD displayed a control-like pattern with all molecular forms of acyl-CoA oxidase (72, 52 and 20 kDa) present, whereas in the W78/515 cell line only the 72 kDa component could be visualised. The GM3605 cell line was intermediate in this respect.
authors.affiliation: E.C. Slater Institute for Biochemical Research, University of Amsterdam, The Netherlands.
authors.forename: E A
authors.initials: EA
authors.lastname: Wiemer
authors.forename: M
authors.initials: M
authors.lastname: Out
authors.forename: A
authors.initials: A
authors.lastname: Schelen
authors.forename: R J
authors.initials: RJ
authors.lastname: Wanders
authors.forename: R B
authors.initials: RB
authors.lastname: Schutgens
authors.forename: H
authors.initials: H
authors.lastname: Van den Bosch
authors.forename: J M
authors.initials: JM
authors.lastname: Tager
mesh_terms: D000101:Acetyl-CoA C-Acetyltransferase / Q000032:analysis; D042962:Acyl-CoA Oxidase; D000217:Acyltransferases / Q000032:analysis / Q000378:metabolism; D002374:Catalase / Q000032:analysis / Q000378:metabolism; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007700:Kinetics; D008830:Microbodies / Q000201:enzymology* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010088:Oxidoreductases / Q000032:analysis; D010641:Phenotype; D010875:Pipecolic Acids / Q000097:blood*; D012035:Refsum Disease / Q000201:enzymology* / Q000378:metabolism; D015211:Zellweger Syndrome / Q000201:enzymology* / Q000378:metabolism
chemical_list: D010875:Pipecolic Acids; D010088:Oxidoreductases; D002374:Catalase; D042962:Acyl-CoA Oxidase; D000217:Acyltransferases; C020051:glycerone-phosphate O-acyltransferase; D000101:Acetyl-CoA C-Acetyltransferase