title: Visualization of mitochondria with green fluorescent protein in cultured fibroblasts from patients with mitochondrial diseases.
abstract: cDNAs for green fluorescent protein (GFP) and for a GFP fusion protein containing the presequence of human ornithine transcarbamylase (pOTC-GFP) were transfected into cultured human fibroblasts. GFP cDNA gave diffuse fluorescence throughout the cytoplasm and the nucleus, whereas pOTC-GFP cDNA gave mitochondria-associated fluorescence. Fluorescent mitochondrial structures could be classified into five patterns: thread-like mitochondria, fine thread-like ones, rod-like ones, granular ones, and granular ones with weak cytosolic fluorescence. pOTC-GFP mutants resulted in a loss of mitochondrial fluorescence and an appearance of weak fluorescence throughout the cytoplasm. pOTC-GFP cDNA was transfected into fibroblasts from patients with various mitochondrial diseases. Higher ratios of fibroblasts with granular mitochondria and those with fine thread-like ones were observed in a patient with Reye's syndrome and a patient with Kearns-Sayre syndrome. Weak cytosolic fluorescence was sometimes observed in fibroblasts from these patients. This method will be useful to analyze mitochondrial structural alterations and disorders of mitochondrial protein import.
authors.affiliation: Department of Pediatrics, Chiba University, School of Medicine, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Kanazawa
authors.forename: M
authors.initials: M
authors.lastname: Yano
authors.forename: C
authors.initials: C
authors.lastname: Namchai
authors.forename: S
authors.initials: S
authors.lastname: Yamamoto
authors.forename: A
authors.initials: A
authors.lastname: Ohtake
authors.forename: M
authors.initials: M
authors.lastname: Takayanagi
authors.forename: M
authors.initials: M
authors.lastname: Mori
authors.forename: H
authors.initials: H
authors.lastname: Niimi
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics; D005347:Fibroblasts / Q000737:chemistry / Q000145:classification / Q000473:pathology; D049452:Green Fluorescent Proteins; D006801:Humans; D007625:Kearns-Sayre Syndrome / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D008164:Luminescent Proteins / Q000032:analysis* / Q000235:genetics; D008661:Metabolism, Inborn Errors / Q000201:enzymology / Q000378:metabolism / Q000473:pathology*; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000201:enzymology / Q000235:genetics / Q000473:pathology*; D009954:Ornithine Carbamoyltransferase / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics; D012202:Reye Syndrome / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D037401:Scyphozoa
chemical_list: D018076:DNA, Complementary; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D009954:Ornithine Carbamoyltransferase