|News and Events||About COST Action MitoEAGLE||Working Groups||MitoEAGLE Summit 2020 Obergurgl AT||MitoEAGLE Preprint States and rates||Short-Term Scientific Missions||Inclusiveness Target Countries||Management Committee||Members|
- COST Action CA15203 MitoEAGLE
Evolution-Age-Gender-Lifestyle-Environment: mitochondrial fitness mapping
- COST Action CA15203 MitoEAGLE
MitoPedia topics: EAGLE
|Name||Ledo Ana Margarida, PhD|
Redox Biology in Health and Disease Group
Faculdade de Medicina, Polo I, Piso I
|Address||Rua Larga, 3004-504|
|O2k-Network Lab||PT Coimbra Laranjinha J|
|Gnaiger 2019 MitoFit Preprint Arch||2019||Gnaiger E, Aasander Frostner E, Abdul Karim N, Abdel-Rahman EA, Abumrad NA, Acuna-Castroviejo D, Adiele RC, et al (2019) Mitochondrial respiratory states and rates. MitoFit Preprint Arch doi:10.26124/mitofit:190001.v6.|
|Dias 2018 Anal Biochem||2018||Dias C, Lourenco CF, Barbosa RM, Laranjinha J, Ledo A (2018) Analysis of respiratory capacity in brain tissue preparations: high-resolution respirometry for intact hippocampal slices. Anal Biochem 551:43-50.|
|Dias 2016 Neurobiol Aging||2016||Dias C, Lourenço CF, Ferreiro E, Barbosa RM, Laranjinha J, Ledo A (2016) Age-dependent changes in the glutamate-nitric oxide pathway in the hippocampus of the triple transgenic model of Alzheimer's disease: implications for neurometabolic regulation. Neurobiol Aging 46:84-95.|
|Lourenco 2015 Front Aging Neurosci||2015||Lourenço CF, Ledo A, Dias C, Barbosa RM, Laranjinha J (2015) Neurovascular and neurometabolic derailment in aging and Alzheimer's disease. Front Aging Neurosci 7:103.|
|Dias 2019 MiPschool Coimbra||2019|
MitoEAGLE Short-Term Scientific Mission
- Work Plan summary
- MitoEAGLE STSM “ University of Szeged, 24-30 March 2018 Title: Respirometric Protocols for Intact Tissue Preparations - Regulation of mitochondrial respiration by bioactive gases Aim and motivation: One of my main research interests over the years has been to better understand how nitric oxide (NO) may act as a master regulator of the neurometabolic-neurovascular coupling axis in the hippocampus, and how changes in NO bioactivity may have implications for the molecular mechanisms leading to degeneration and disease in the aging brain. We have performed several studies using carbon fiber microelectrodes to measure both NO and O2 in hippocampal slice preparations as well as in vivo in anesthetized rats. More recently we have also performed O2 recordings in the brain of free-behaving rodents using chronically implanted Pt microelectrode arrays. To an extent this has allowed us to better understand the complex interplay between the two bioactive gas molecules. Considering the established knowledge of compromised bioenergetics function in both aging and degeneration, we aimed to evaluate mitochondrial dysfunction in discrete brain structures such as the hippocampus. Two factors have lead us to design respirometric protocols using intact brain slices: firstly, limited tissue volume; secondly, the desire to retail tissue cytoarchitecture, mitochondrial physiology, intercellular communication as neuron-astrocyte metabolite shuttling. The Oroboros system is unique in allowing this type of studies using intact tissue preparation, making it possible to evaluate mitochondrial function in structures for which isolation of mitochondria or cells is not feasible. The fact that we have such an equipment available has led to several collaboration proposals from other groups looking to assess mitochondrial function in biopsies from human subjects as well as diverse tissue samples from animal models of disease (obesity, diabetes, etc.). Thus far we have had the chance to use, besides hippocampal slices, pericardiac adipose tissue, perivascular adipose tissue as well as gastric and hepatic biopsies. We have come to realize, that each tissue type requires protocol adjustments and optimizations. Herein lies the aim and motivation to embark on this STSM to University of Szeged, Institute of Surgical Research “ the possibility to contact with other research groups with expertise in performing respirometry studies in intact and permeabilized tissue preparations.
- Also relevant is the fact that this particular group has experience and knowledge regarding NO and it related reactive nitrogen species (namely peroxynitrite), as well as nitrite and nitrate. Proposed Contributions to the objectives of MitoEAGLE One of the main objectives of the MitoEAGLE network is to improve our current understanding of mitochondrial function in health and disease. Having ourselves designed protocols to evaluate mitochondrial function in intact hippocampal slices, this STSM may contribute to the objectives of MitoEAGLE as discussion and comparative analysis of protocols with others will allow improvement of quality control and allow for innovation in the field of mitochondrial respiratory physiology. The possibility to discuss our protocols within this network of expert researchers will help decrease the lack of uniformity and consensus on standard operating procedures in the design and implementation of research protocols involving mitochondrial physiology. Furthermore, we will join our data to the already available pool of data collected from other groups working in mitochondrial physiology in health and disease. Also relevant is the standardization of data presentation and terminology, critical to allow understanding of data being produced worldwide and derived from diverse tissue preparations. As one of the few groups using intact brain tissue preparations, we can contribute with our unique perspective and needs not only in terms of protocol design, but also data management and presentation. Techniques to be Explored: Isolation of biopsies from the gastrointestinal (GI) tract for high resolution respirometric assays; High-resolution respirometric SUIT protocols for GI tract biopsies:
- permeabilization, tissue preservation, substrate-uncoupler-inhibitor-titrations, evaluation of mitochondrial complex activity in GI-tract biopsies/slices, regulation of mitochondrial respiration by bioactive gases and associated metabolites (NO/NO2-/ONOO-).
- Detailed Plan March 24: Travel from Coimbra, Portugal to Szeged, Hungary March 25: Business meeting with Andras Meszaros and Erich Gnaiger “ discussion on protocols and standard operating procedures, data acquisition and presentation, presentation of our models, approaches, problems and solutions.
- March 26-27: Attending O2k Workshop in Hypoxia (presenting short talk on the 27, entitled “Age-dependent changes in the glutamate-nitric oxide pathway in the hippocampus of the triple transgenic model of Alzheimer’s disease: implications on mitochondrial function”) March 28-29: In the lab (Institute of Surgical Research), with Dr. Laszlo Juhasz and Prof. Jozsef Kaszaki. Exploring protocols with organ slices, in particular those obtained from the lower GI track (intestine and colon): respirometry studies using the O2k system using intact tissue preparations; regulation of mitochondrial respiration by bioactive gases (ex. NO) and related metabolites (ONOO-).
- March 30 - Travel from Szeged, Hungary to Coimbra, Portugal.
- Work Plan summary