Description du sujet de thèse

The PhD student's scientific objective will be to further characterize the importance of maintaining mitohondrial dynamics and morphology on genome maintenance and cardiac pathophysiology. The student will demonstrate a good level of expertise in the maintenance and follow-up of murine transgenic lines (genotyping, sacrifice and collection of tissues and organs).
The PhD student should have a strong interest in biochemical methods for characterizing mitochondrial bioenergetics (mitochondrial isolation, oxygenation of respiration, native and denaturing electrophoresis, spectroscopy and fluorometry), cardiac metabolism (metabolomics and perfusion of isolated organs), and the structure of the mitochondrial compartment (confocal and electron microscopy).
The PhD student will also develop approaches for characterizing mitochondrial function in situ on a perfused beating heart (working mode and Langendorff), using a system of optical fibers coupled to a fluorometer.

Activities
- Genotyping and lineage monitoring, sacrifice and tissue collection
- Metabolic biochemistry experiments (isolation of mitochondria from tissue or cultured cells, measurement of oxygen consumption flux by high-resolution O2K oroboros oxygraph, measurement of enzymatic activities by spectrophotometry or fluorimetry, native and denaturing electrophoresis).
- Extraction and quenching of tissue or cardiac perfusion effluent for metabolomic analysis. Cardiac physiological activity measurements (MVO2, ECG, mechanical activity, calcium transients, autofluorescence of intrinsic and extrinsic chromophores).
- Isolation of ventricular and atrial myocytes for functional analysis (oxygenography, fluorometry, confocal microscopy, FACS) and structural analysis of mitochondrial membrane arrangement (confocal and electron microscopy).

Expected skills
- Animal experimentation operator diploma desirable (training available on the job)
- Good command of biochemical measurements (high-resolution oxygenography, enzymatic activities, denaturing and native protein gels, notions of cardiovascular pathophysiology, spectro and fluorimetric approaches)
- Good command of English (level B2 to C1 of the European Framework of Reference for Languages)

Contexte de travail

The 3-year PhD contract is part of an ANR 'DynaMitoPatho' co-financing and 'MetabOptic' regional project in collaboration with Dr. Pierre Bon's laboratory (XLILM Research Center, Limoges). Recruitment will take place within the BioDynaMit laboratory located at IBGC-UMR5095.
The laboratory has a very high level of expertise in cellular and mitochondrial energy metabolism. We have a wide range of equipment and animal models for studying the importance of mitochondrial metabolism in metabolic maintenance and cardiac pathophysiology.
The person recruited will be placed under the responsibility of Dr. Philippe Pasdois, an expert in cardiac energetics and the study of mitochondrial function on isolated organs, in close collaboration with Dr. Arnaud Mourier, an expert in mitochodrial genetic diseases and murine transgenic models.