Lysosomal Metabolism Core

Coordinators:

Prof. Ivan Conte

Dr. Giamundo Giuliana

email:

ivan.conte@unina.it

giamundo.giuliana@unina.it

The Department of Biology at University of Naples Federico II hosts the Lysosomal Metabolism Core which provides resources for research projects and teaching classes for researchers and Ph.D. students that are interested to understand the mechanistic link between lysosomal activity and the cell homeostasis in physiological or pathological states. Lysosomes satisfy basic requirements for nutrient sensing, utilization and recycling through macromolecular breakdown through the synchronized interplay between hydrolytic enzymes, nutrient transporters and signaling factors, which together enable metabolic coordination with other organelles and coordinate programs for growth, repair and stress adaptation. Furthermore, it is increasingly recognized that acquired lysosomal dysfunction is a hallmark of highly prevalent diseases such as lysosomal storage disorders, metabolic diseases, cancer, age-related diseases, neurodegenerative disorders.

Thus, the Core provides an opportunity to explore common scientific themes and collaborate on research combining the molecular and cellular mechanisms for the lysosomal metabolism with a wide range of studies.

The lysosomal metabolism core holds the following areas:

High-resolution confocal imaging of lysosomes

• Lysosomal cargoes internalization and degradation
• Lysosomal contact sites and fusion:
• Identification of inter-organellar associations in fixed and living cells
• Lysosomal activity:
• Assessment of lysosomal enzymatic activities
• Analysis of lysosomal hydrolase activity:
• Ratiometric assessment of lysosomal acidification and pH
• Lysosomal biogenesis
• Lysosomal positioning and motility
• Lysosomal exocytosis

ACKNOWLEDGEMENTS

All the users and collaborators of the Core Cell Metabolism Research are obligated to acknowledge the core in publications: “The authors acknowledge the Lysosomal Metabolism Core at Department of Biology of University of Naples Federico II.

Mitochondrial Metabolism Core

Coordinator:

Prof. Susanna Iossa

email: susanna.iossa@unina.it

The Department of Biology at University of Naples Federico II hosts the Mitochondrial Metabolism Core which provides resources for research projects and teaching classes for researchers and Ph.D students that are interested to deepen the role of metabolic and mitochondrial functions in physiological and/or disease states. Today, mitochondria and metabolism studies continue to fascinate us, since the alterations of mitochondrial function and cell metabolism control life span in multiple model organisms. Furthermore, it is increasingly recognized that acquired mitochondrial dysfunction is a hallmark of highly prevalent diseases such as obesity/diabetes, cancer, cardiovascular and neurodegenerative diseases.

The Mitochondrial Metabolism Core, the only Italian organization that offers high resolution respirometry to internal researchers, as well as national and international collaborators, and industrial partners, has been established since 2022 to support the advancement of our understanding of the basic biological mechanisms that regulate metabolism, and how they are dysregulated in disorders such as obesity, diabetes, neurodegenerative diseases, and cancer. While these may seem to be divergent themes, they share common root causes in disordered energy balance, which can affect many biological systems. Thus, the Core provides an opportunity to explore common scientific themes and collaborate on research combining the molecular and cellular mechanisms of metabolism with a wide range of physiological and behavioral studies.

The Mitochondrial metabolism core holds the following instruments:

Equipment:

Oroboros O2K- is a state-of-the art respirometer for quantitative high-resolution respirometry and comprehensive OXPHOS analysis and simultaneous real-time monitoring of mitochondrial membrane potential in isolated mitochondria, permeabilized tissues (e.g. skeletal muscle fibres), intact cells, permeabilised cells and tissue homogenates.

Research Areas:

• Cell Metabolism, in particular fatty acid metabolism
• Mitochondrial physiology, with a special focus on bioenergetics processes
• Mitochondria in cardiac, skeletal muscle and liver/metabolic diseases
• Brain mitochondrial function and its role in neurodegeneration – in collaboration with Luisa Cigliano

Coordinators : Susanna Iossa (susanna.iossa@unina.it) , Raffaella Crescenzo (raffaella.crescenzo@unina.it), Arianna Mazzoli (arianna.mazzoli@unina.it)

Equipment:

The Seahorse XFp24 instrument performs analyses of cellular metabolic function in cultured cells and ex-vivo samples through the measurements of the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR), key indicators of mitochondrial respiration and glycolysis, as well as ATP production rate

Research Areas:

• Cell Metabolism, in particular fatty acid metabolism
• Mitochondrial physiology, with a special focus on bioenergetics processes
• Mitochondria in cardiac, skeletal muscle and liver/metabolic diseases
• Brain synaptosomal mitochondrial function and its role in neuro-disorders – in collaboration with Marianna Crispino

Coordinators: Maria Pina Mollica (mpmollic@unina.it), Giovanna Trinchese (giovanna.trinchese@unina.it)

Equipment:

The Hansatech Instruments Oxygen Electrode Measurement System enables the evaluation of oxygen uptake in various applications for the quantification of mitochondrial respiration in isolated organelles, tissue homogenate, and cells as well as for the analysis of oxidative phosphorylation. It includes an integral system for measuring pH and other ion-selective electrode signals, for monitoring mitochondrial membrane potential.

Research Areas:

• Cell Metabolism, in particular fatty acid metabolism
• Mitochondrial physiology, with a special focus on bioenergetics processes
• Mitochondria in cardiac, skeletal muscle and liver/metabolic diseases

Coordinator: Assunta Lombardi (assunta.lombardi@unina.it)

Research Areas:

• Role of mitochondria in ROS production and ROS scavenging in conditions of altered oxidative balance.

Coordinator: Paola Venditti (paola.venditti@unina.it)

ACKNOWLEDGEMENTS

All the users and collaborators of the Core Cell Metabolism Research are obligated to acknowledge the core in publications: “The authors acknowledge the Mitochondrial Metabolism Core at Department of Biology of University of Naples Federico II.