Talk:ESCI 2016 Paris FR
Mitochondrial respiratory control patterns of oxidative phosphorylation in human prostate tissue and cell lines
Bernd Schöpf1, Georg Schäfer2,3, Anja Weber2, Iris E. Eder2, Helmut Klocker2, Erich Gnaiger4
- Division of Genetic Epidemiology, Department of Medical Genetics, Molecular and Clinical Pharmacology; Medical University of Innsbruck, Schöpfstrasse 41, A-6020 Innsbruck, Austria, E: email@example.com
- Division of Experimental Urology, Department of Urology, Medical University of Innsbruck, Anichstraße 35, A-6020 Innsbruck, Austria, E: firstname.lastname@example.org, email@example.com, firstname.lastname@example.org, email@example.com
- Department of Pathology, Medical University of Innsbruck, Müllerstrasse 44, A-6020 Innsbruck, Austria, E: firstname.lastname@example.org
- Department of General and Transplant Surgery, D. Swarovski Research Laboratory, Medical University of Innsbruck, Innrain 66/6, A-6020 Innsbruck, Austria
Altered mitochondrial metabolism plays a pivotal role in the development and progression of various diseases, including cancer. Cell lines are frequently used as models to study mitochondrial dysfunction. However, little is known about mitochondrial respiratory control in cultured cells compared to the primary tissue of origin. The analysis of mitochondrial function in tissue is a significant analytical challenge due to limited sample quantities. We have developed a method for assessment of oxidative phosphorylation in prostate tissue biopsies of only 2 mg wet weight using high-resolution respirometry. Various substrate-uncoupler-inhibitor-titration protocols were evaluated and applied to measure the activity of different segments of the electron transfer system in mechanically permeabilized benign prostatic tissue biopsy samples. Additionally, the widely used prostate epithelial cell line RWPE1 and immortalized prostate fibroblasts, representing the major cell types in prostate tissue, were analyzed after chemical permeabilization to compare cell line-based models and the tissue of origin. Our results show that mechanical treatment without addition of permeabilization agents or isolation of mitochondria constitutes a reliable preparation method for OXPHOS analysis in small amounts of prostatic tissue typically obtained by prostate biopsy. The cell lines represented the bioenergetic properties of fresh tissue to a limited extent only. However, the flexibility of cell culture models allowing control of metabolic conditions in combination with tissue samples may prove to be powerful tools for bioenergetic metabolism studies in health and disease as well as metabolic biomarker discovery.
Accommodation & travel arrangements
- Staying at: Port Royal Hôtel, Apr 27 to May 01.
- Travel arrangements in process.
- Freistellungsantrag to be submitted.
Laner Verena 18:20, 28 January 2016 (CET)