title: Involvement of the mitochondrial compartment in human NCL fibroblasts.
abstract: Neuronal ceroid lipofuscinosis (NCL) are a group of progressive neurodegenerative disorders of childhood, characterized by the endo-lysosomal storage of autofluorescent material. Impaired mitochondrial function is often associated with neurodegeneration, possibly related to the apoptotic cascade. In this study we investigated the possible effects of lysosomal accumulation on the mitochondrial compartment in the fibroblasts of two NCL forms, CLN1 and CLN6. Fragmented mitochondrial reticulum was observed in all cells by using the intravital fluorescent marker Mitotracker, mainly in the perinuclear region. This was also associated with intense signal from the lysosomal markers Lysotracker and LAMP2. Likewise, mitochondria appeared to be reduced in number and shifted to the cell periphery by electron microscopy; moreover the mitochondrial markers VDCA and COX IV were reduced following quantitative Western blot analysis. Whilst there was no evidence of increased cell death under basal condition, we observed a significant increase in apoptotic nuclei following Staurosporine treatment in CLN1 cells only. In conclusion, the mitochondrial compartment is affected in NCL fibroblasts invitro, and CLN1 cells seem to be more vulnerable to the negative effects of stressed mitochondrial membrane than CLN6 cells.
authors.affiliation: Department of Neurological, Psychological, Morphological and Motor Sciences, Divisions of Neurology (Child Neurology) and Neuropathology, University of Verona Medical School, Verona, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Pezzini
authors.forename: Floriana
authors.initials: F
authors.lastname: Gismondi
authors.forename: Alessandra
authors.initials: A
authors.lastname: Tessa
authors.forename: Paola
authors.initials: P
authors.lastname: Tonin
authors.forename: Rosalba
authors.initials: R
authors.lastname: Carrozzo
authors.forename: Sara E
authors.initials: SE
authors.lastname: Mole
authors.forename: Filippo M
authors.initials: FM
authors.lastname: Santorelli
authors.forename: Alessandro
authors.initials: A
authors.lastname: Simonati
mesh_terms: D000588:Amines / Q000737:chemistry; D015415:Biomarkers / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D002478:Cells, Cultured; D005347:Fibroblasts / Q000187:drug effects / Q000648:ultrastructure; D006801:Humans; D052119:Lysosomal-Associated Membrane Protein 2; D051907:Lysosomal Membrane Proteins / Q000378:metabolism; D008247:Lysosomes / Q000648:ultrastructure; D008565:Membrane Proteins / Q000378:metabolism; D008928:Mitochondria / Q000648:ultrastructure*; D009472:Neuronal Ceroid-Lipofuscinoses / Q000473:pathology*; D019311:Staurosporine / Q000494:pharmacology; D013869:Thiolester Hydrolases
chemical_list: D000588:Amines; D015415:Biomarkers; C447500:CLN6 protein, human; C497811:LAMP2 protein, human; D052119:Lysosomal-Associated Membrane Protein 2; D051907:Lysosomal Membrane Proteins; D008565:Membrane Proteins; C493330:Red DND-99; D013869:Thiolester Hydrolases; C416054:PPT1 protein, human; D053148:Caspase 3; D019311:Staurosporine