Joshi 2022 Abstract Bioblast

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P02.
Joshi Saurabh
Joshi Saurabh, Hater F, Eirich J, Palovaara J, Ellinghaus H, Heinkow P, Callenius H, Peter A, Schweser O, Kubitschke M, Madduri MK, Mathew AJ, Ciacchi LC, Kirstein J, Maedler K, Masseck OA, Finkemeier I, Radmacher M, Groß-Hardt R (2022) A versatile mitochondria isolation- and analysis-pipeline generates 3D nano-topographies and mechano-physical surface maps of single organelles. Bioblast 2022: BEC Inaugural Conference. In: https://doi.org/10.26124/bec:2022-0001

Link: Bioblast 2022: BEC Inaugural Conference

Joshi Saurabh, Hater F, Eirich J, Palovaara J, Ellinghaus H, Heinkow P, Callenius H, Peter A, Schweser O, Kubitschke M, Madduri MK, Mathew AJ, Ciacchi LC, Kirstein J, Maedler K, Masseck OA, Finkemeier I, Radmacher M, Groß-Hardt R (2022)

Event: Bioblast 2022

Living eukaryotic cells typically contain large quantities of highly dynamic mitochondria, which sustain the cells’ energy and redox homeostasis. Growing evidence suggests that mitochondria can functionally differ among but also within cells. The extent and biological significance of mitochondrial diversity is still largely unexplored, due to technical limitations that hamper profiling of individual organelles. Previous measurements of the cell’s interior have shown that membrane-bound compartments respond to metabolic manipulation by changes in their surface stiffness, suggesting that mechano-physical properties are a valuable readout of mitochondrial function. We here present the establishment of a robust multi-step analysis pipeline that allows one to profile mechano-physical properties of single mitochondria at the nanoscale using Atomic Force Microscopy (AFM) [1]. Firstly, we developed a rapid cell-type specific isolation protocol (mRACE), which selectively functionalizes mitochondria with biotin, facilitating isolation by streptavidin decorated microbeads. We established the technique for human and rat cell cultures, the invertebrate Caenorhabditis elegans, and the model plant Arabidopsis thaliana. Based on this versatile tool, we detected diversity of mitochondrially associated proteins among different tissues, reflecting the trophic condition of the source material. Secondly, a rapid filtration-based mitochondria isolation protocol was established, which was combined with mRACE. Lastly, we established an AFM analysis platform, which generates 3D maps of the nano-topography and mechano-physical properties of individual mitochondria. The comparison of mitochondria with each other revealed an unprecedented diversity in their mechano-physical properties and suggests that shape is not the sole determining parameter for mitochondrial outer membrane stiffness. We expect our results to not only introduce a new dimension for basic mitochondrial research, but in addition to open the door for the exploitation of individual mitochondria for diagnostic characterization [1].

  1. Saurabh J, Hater F, Eirich J, Palovaara J, Ellinghaus H, Heinkow P, Callenius H, Peter A, Schweser O, Kubitschke M, Madduri MK, Mathew AJ, Ciacchi LC, Kirstein J, Maedler K, Masseck OA, Finkemeier I, Radmacher M, Groß-Hardt R (2021) A versatile mitochondria isolation- and analysis-pipeline generates 3D nano-topographies and mechano-physical surface maps of single organelles. https://doi.org/10.1101/2021.10.31.466655 - The present abstract is a copy from the preprint.


Bioblast editor: Gnaiger E


Affiliations and support

Saurabh J§1, Hater F§1, Eirich J2, Palovaara J1, Ellinghaus H1, Heinkow P2, Callenius H1, Peter A1, Schweser O1, Kubitschke M1, Madduri MK1, Mathew AJ1, Ciacchi LC4, Kirstein J1&, Maedler K1&, Masseck OA1&, Finkemeier I2, Radmacher M3*, Groß-Hardt R*1
  1. University of Bremen, Centre for Biomolecular Interactions, 28359 Bremen, Germany
  2. Plant Physiology, Institute of Plant Biology and Biotechnology, University of Muenster, Schlossplatz 7, DE-48149 Muenster, Germany
  3. University of Bremen, Institute of Biophysics, Otto-Hahn Allee 1, D-28359 Bremen, Germany
  4. University of Bremen, Hybrid Materials Interfaces Group, Faculty of Production Engineering and Centre for Environmental Research and Sustainable Technology (UFT), 28359 Bremen, Germany
§ These authors contributed equally to this work
& These authors contributed equally to this work
* Correspondence to: gross-hardt@uni-bremen.de, radmacher@uni-bremen.de
This work is supported by the European Research Council (ERC Consolidator Grant "bi-BLOCK" ID. 646644 to Rita Groß-Hardt).

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Preparation: Isolated mitochondria 



Event: Poster