Mitochondrial oxidative phosphorylation and energetic status are reflected by morphology of mitochondrial network in INS-1E and HEP-G2 cells viewed by 4Pi microscopy.pdfVIP

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Mitochondrial oxidative phosphorylation and energetic status are reflected by morphology of mitochondrial network in INS-1E and HEP-G2 cells viewed by 4Pi microscopy.pdf

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Mitochondrial oxidative phosphorylation and energetic status are reflected by morphology of mitochondrial network in INS-1E and HEP-G2 cells viewed by 4Pi microscopy.pdf

Biochimica et Biophysica Acta 1777 (2008) 834–846 Contents lists available at ScienceDirect Biochimica et Biophysica Acta j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / b b a b i o Mitochondrial oxidative phosphorylation and energetic status are re?ected by morphology of mitochondrial network in INS-1E and HEP-G2 cells viewed by 4Pi microscopy Lydie Plecitá-Hlavatá a, Mark Lessard b, Jitka ?antorová a, Joerg Bewersdorf b, Petr Je?ek a,? a Department of Membrane Transport Biophysics, No. 75, Institute of Physiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220 Prague 4, Czech Republic b Institute for Molecular Biophysics, The Jackson Laboratory, Bar Harbor, Maine, USA ARTICLE INFO Article history: Received 30 January 2008 Received in revised form 31 March 2008 Accepted 1 April 2008 Available online 10 April 2008 Keywords: 3D morphology of mitochondrial network 4Pi microscopy Uncoupling Oxidative phosphorylation Insulinoma INS-1E cell Hepatocellular carcinoma HEP-G2 cell ABSTRACT Mitochondria in numerous cell types, especially in cultured cells, form a reticular network undergoing constant fusion and ?ssion. The three dimensional (3D) morphology of these networks however has not been studied in detail to our knowledge. We have investigated insulinoma INS-1E and hepatocellular carcinoma HEP-G2 cells transfected with mitochondria-addressed GFP. Using 4Pi microscopy, 3D morphology changes responding to decreased oxidative phosphorylation and/or energetic status could be observed in these cells at an unprecedented 100 nm level of detail. In INS-1E cells cultivated at 11 mM glucose, the mitoreticulum appears predominantly as one interconnected mitochondrion with a nearly constant 262 ± 26 nm tubule diameter. If cultured at 5 mM glucose, INS-1E cells show 311 ± 36 nm tubules coexisting with numerous ?at cisternae. Similar interconnected 284 ± 38 nm and 417 ± 110 nm tubules were found in HEP-G2 cells cultivated at 5 mM and hyp