These authors contributed equally to this work.
Version of Record online: 24 JUN 2011
Copyright © 2011 American Association for the Study of Liver Diseases
Volume 54, Issue 1, pages 296–306, July 2011
How to Cite
Guerra, M. T., Fonseca, E. A., Melo, F. M., Andrade, V. A., Aguiar, C. J., Andrade, L. M., Pinheiro, A. C. N., Casteluber, M. C. F., Resende, R. R., Pinto, M. C. X., Fernandes, S. O. A., Cardoso, V. N., Souza-Fagundes, E. M., Menezes, G. B., de Paula, A. M., Nathanson, M. H. and de Fátima Leite, M. (2011), Mitochondrial calcium regulates rat liver regeneration through the modulation of apoptosis. Hepatology, 54: 296–306. doi: 10.1002/hep.24367
Potential conflict of interest: Nothing to report.
This work was supported by grants from the Howard Hughes Medical Institute (to Maria de Fátima Leite), Conselho Nacional de Desenvolvimento Científico e Tecnológico (to Maria de Fátima Leite, Rodrigo R. Resende, Gustavo B. Menezes, Valbert N. Cardoso, and Ana M. de Paula), Fundação de Amparo à Pesquisa do Estado de Minas Gerais (to Maria de Fátima Leite, Rodrigo R. Resende, Gustavo B. Menezes, Valbert N. Cardoso, and Ana M. de Paula), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (to Mateus T. Guerra, Viviane A. Andrade, Marisa F. Casteluber, and Rodrigo R. Resende), Instituto Nacional de Ciência e Tecnologia (to Carla J. Aguiar), and the National Institutes of Health (DK57751, DK45710, and DK34989 to Michael H. Nathanson).
- Issue online: 24 JUN 2011
- Version of Record online: 24 JUN 2011
- Accepted manuscript online: 18 APR 2011 08:31AM EST
- Manuscript Accepted: 5 APR 2011
- Manuscript Received: 4 JAN 2011
Additional Supporting Information may be found in the online version of this article.
|HEP_24367_sm_suppinfofig1.tif||118K||Supporting Figure 1. Targeting PV to mitochondria does not induce cell proliferation. Control SKHep1 cells and cells transfected with MITO-GFP or PV MITO GFP were treated for 6 hours with or without 300 nM STA. Expression of PV in mitochondria did not alter cell proliferation. STA treatment induced cell death in 20.6±6.2% of MITO-GFP cells. Under the same conditions, PV expression prevented cell death, although it did not induce cell proliferation. Results represent the mean±S.E.M. of 3 separate experiments.|
|HEP_24367_sm_suppinfofig2.tif||1403K||Supporting Figure 2. Targeting PV expression in rat liver. Three sequential planes of an intravital confocal Z-stack (160 μM in 2 μM increments) of a rat hepatic lobule 2 days after tail vein injection of Ad-PV-MITO-GFP. PV is shown in green.|
|HEP_24367_sm_suppinfofig3.tif||471K||Supporting Figure 3. Intravital confocal images of rat hepatic lobule and intestine. Upper panels show representative intravital confocal images of the liver of a rat 2 days after tail vein injection of saline solution. No fluorescence was observed in the liver under the same confocal settings used to capture images from livers expressing PV-GFP. Lower panels show representative confocal images of intestine of an Ad-PV-MITO-GFP- injected rat (3 × 109 pfu/kg), indicating absence of PV expression (green) in this organ.|
|HEP_24367_sm_suppinfofig4.tif||4127K||Supporting Figure 4. Evaluation of inflammatory infiltrate during liver regeneration. A. MPO activity was used to evaluate neutrophil infiltration in sham, PH or PH + Ad-PV-MITO livers. Inflammation was similarly increased at day 1 in PH and PH + Ad-PV-MITO relative to sham (p<0.02 by one-way ANOVA) but returned to control levels after day 2 post-surgery. B. Representative hematoxylin-eosin staining shows similar liver architecture with minimal inflammatory infiltrate in both PH and PH + Ad-PV-MITO during 3 days of liver regeneration.|
|HEP_24367_sm_suppinfofig5.tif||1257K||Supporting Figure 5. Liver morphology after PH. A. Liver body weight index 7 days after PH demonstrates no difference in liver mass between sham-operated and animals subjected to PH with or with injection of Ad-PV-MITO-GFP. B. Hematoxylin-eosin staining of representative liver sections from sham-operated rats, rats after PH, and rats infected with Ad-PV-MITO-GFP one day prior to PH. Analysis was performed 7 days after PH. No identifiable morphological alteration was observed among the livers (n=3 rats per condition).|
|HEP_24367_sm_suppinfofig6.tif||147K||Supporting Figure 6. Liver function. A-F. Serum albumin, bilirubin, AST, ALT, and ALP were evaluated in samples from sham operated rats, rats after PH, and rats infected with Ad-PV MITO-GFP. There were no significant changes in any of the biochemical parameters among the groups (n=3 experiments).|
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