University of Crete

5 ARTICLES PUBLISHED IN JoVE

Medicine

In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice

Evandro F. Fang^1,6, Konstantinos Palikaras², Nuo Sun³, Elayne M. Fivenson¹, Ryan D. Spangler⁴, Jesse S. Kerr¹, Stephanie A. Cordonnier¹, Yujun Hou¹, Eszter Dombi⁵, Henok Kassahun⁶, Nektarios Tavernarakis^2,7, Joanna Poulton⁵, Hilde Nilsen⁶, Vilhelm A. Bohr^1,8

¹Laboratory of Molecular Gerontology, National Institute on Aging, National Institutes of Health, ²Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology - Hellas, ³Center for Molecular Medicine, National Heart Lung and Blood Institute, National Institutes of Health, ⁴Laboratory of Neurosciences, National Institute on Aging, National Institutes of Health, ⁵Nuffield Department of Obstetrics and Gynaecology, University of Oxford, ⁶Department of Clinical Molecular Biology, University of Oslo and Akershus University Hospital, ⁷Department of Basic Sciences, Faculty of Medicine, University of Crete, ⁸Danish Center for Healthy Aging, University of Copenhagen

Mitophagy, the process of clearing damaged mitochondria, is necessary for mitochondrial homeostasis and health maintenance. This article presents some of the latest mitophagy detection methods in human cells, Caenorhabditis elegans, and mice.

Biology

Modeling Age-Associated Neurodegenerative Diseases in Caenorhabditis elegans

Konstantinos Palikaras^1,2, Nektarios Tavernarakis^1,2

¹Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, Greece, ²Department of Basic Sciences, Faculty of Medicine, University of Crete, Heraklion, 70013, Crete, Greece

Here, we introduce and describe widely accessible methodologies utilizing some versatile nematode models, including hyperactivated ion channel-induced necrosis and protein aggregate-induced neurotoxicity, to monitor and dissect the cellular and molecular underpinnings of age-associated neurodegenerative diseases.

Biology

Assessment of de novo Protein Synthesis Rates in Caenorhabditis elegans

Margarita Elena Papandreou^*1,2, Konstantinos Palikaras^*1,2, Nektarios Tavernarakis^1,2

Here, we introduce and describe a nonradioactive and noninvasive method to assess de novo protein synthesis in vivo, utilizing the nematode Caenorhabditis elegans and fluorescence recovery after photobleaching (FRAP). This method can be combined with genetic and/or pharmacological screens to identify novel modulators of protein synthesis.

Chemistry

Versatile Dual-Inlet Sample Introduction System for Multi-Mode Single Particle Inductively Coupled Plasma Mass Spectrometry Analysis and Validation

Daniel Rosenkranz¹, Fabian L. Kriegel¹, Emmanouil Mavrakis², Spiros A. Pergantis², Philipp Reichardt¹, Jutta Tentschert¹, Norbert Jakubowski⁴, Peter Laux¹, Ulrich Panne³, Andreas Luch¹

¹Department of Chemical and Product Safety, German Federal Institute for Risk Assessment (BfR), ²Environmental Chemical Processes Laboratory, Department of Chemistry, University of Crete, ³Federal Institute for Materials Research and Testing (BAM), ⁴SPETEC GmbH

Here we provide a protocol for the use of a dual-inlet system for single particle inductively coupled mass spectrometry which allows for a standard independent nanoparticle characterization.

Genetics

Using the GAL4-UAS System for Functional Genetics in Anopheles gambiae

Beth Crawford Poulton¹, Fraser Colman¹, Amalia Anthousi^1,2,3, Linda Grigoraki¹, Adriana Adolfi¹, Amy Lynd¹, Gareth John Lycett¹

¹Department of Vector Biology, Liverpool School of Tropical Medicine, ²IMBB FORTH, ³Department of Biology, University of Crete

The bipartite GAL4-UAS system is a versatile tool for modification of gene expression in a controlled spatiotemporal manner which permits functional genetic analysis in Anopheles gambiae. The procedures described for using this system are a semi-standardized cloning strategy, sexing and screening of pupae for fluorescent protein markers and embryo fixation.