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Light scattering from intact cells reports oxidative-stress-induced mitochondrial swelling.
Wilson, Jeremy D; Bigelow, Chad E; Calkins, David J; Foster, Thomas H.
Afiliação
  • Wilson JD; Department of Physics and Astronomy, University of Rochester, Rochester, New York 14642, USA.
Biophys J ; 88(4): 2929-38, 2005 Apr.
Article em En | MEDLINE | ID: mdl-15653724
ABSTRACT
Angularly resolved light scattering measurements were performed on suspensions of EMT6 cells and on mitochondria isolated from rabbit liver. Mie theory analysis of the scattering from intact cells indicated that mitochondrial-sized organelles dominated scattering in the range 5-90 degrees . This interpretation was supported by the analysis of scattering from isolated mitochondria. Intact cells were subjected to oxidative stress by photodynamic insult. After 3 h of incubation in the heme precursor aminolevulinic acid hexylester, EMT6 cells accumulated abundant protoporphyrin IX, an endogenous photosensitizer formed in mitochondria. Irradiation of aminolevulinic acid/protoporphyrin IX-sensitized cells with 10 J cm(-2) of 514 nm light led to pronounced changes in angularly resolved light scattering consistent with mitochondrial swelling. Electron microscopy of similarly treated EMT6 cell monolayers showed significant changes in mitochondrial morphology, which included distension of the outer unit membrane and bloating of the internal mitochondrial compartment. Informed by these electron microscopy results, we implemented a coated sphere model to interpret the scattering from intact cells subjected to oxidative stress. The coated sphere interpretation was compatible with the scattering measurements from these cells, whereas simpler Mie theory models based on homogenous swelling were dramatically unsuccessful. Thus, in this system, angularly resolved light scattering reports oxidative-stress-induced changes in mitochondrial morphology.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Biofísica / Estresse Oxidativo / Ácido Aminolevulínico / Microscopia de Polarização / Mitocôndrias Tipo de estudo: Prognostic_studies / Risk_factors_studies Limite: Animals Idioma: En Ano de publicação: 2005 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Biofísica / Estresse Oxidativo / Ácido Aminolevulínico / Microscopia de Polarização / Mitocôndrias Tipo de estudo: Prognostic_studies / Risk_factors_studies Limite: Animals Idioma: En Ano de publicação: 2005 Tipo de documento: Article