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Calzia D.,University of Genoa | Bianchini P.,Italian Institute of Technology | Ravera S.,University of Genoa | Bachi A.,San Raffaele Scientific Institute | And 3 more authors.
Analytical Methods | Year: 2010

The present paper describes a new procedure for labelling living retinas, ex vivo, with fluorescent dyes, in order to image them by microscopy techniques by a procedure that maintains the physiological features of the tissue and its association with the retinal pigmented epithelium (RPE). © 2010 The Royal Society of Chemistry.

Panfoli I.,University of Genoa | Calzia D.,University of Genoa | Bruschi M.,Laboratory on Pathophysiology of Uraemia | Oneto M.,University of Genoa | And 5 more authors.
Cell Biochemistry and Function | Year: 2013

The rod Outer Segment (OS) disc, an organelle devoid of mitochondria, is specialized in phototransduction, a process requiring a continual chemical energy supply. We have shown that OS discs express functional mitochondrial electron transport chains, FoF1-ATP synthase and the tricarboxylic acid cycle enzymes, all mitochondrial features. Here, we focus on oxygen consumption and adenosine triphosphate (ATP) synthesis by OS discs analysing electron transport chain I-III-IV and II-II-IV pathways, supported by reduced nicotinamide adenine dinucleotide and succinate, respectively. Interestingly, respiratory capacity of discs was measurable also in the presence of 3-hydroxy-butyrrate, a typical metabolic substrate for the brain. Data were supported by a two-dimensional electrophoresis analyses conducted as our previous one, but focused to those mitochondrial proteins that are involved in oxidative phosphorylation. Carbonic anhydrase was also found active in OS discs. Moreover, colocalization of Rhodopsin with respiratory complex I and ATP synthase seems a further step in the characterization of some proteins typical of the mitochondrial inner membranes that are expressed in the rod discs. The existence of oxygen utilization in the outer retina, likely supplying ATP for phototransduction, may shed light on some retinal pathologies related to oxidative stress of the outer retina. © 2013 John Wiley & Sons, Ltd.

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