|News and Events||About COST Action MitoEAGLE||Working Groups||MiP/MitoEAGLE Traning School 2019 Coimbra PT||MitoEAGLE Task Group 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
|Institution||Latvian Institute of Organic Sythesis, LV|
|Address||Aizkraukles 21, LV1006|
|Gnaiger 2019 MitoFit Preprint Arch||2019||Gnaiger E, Aasander Frostner E, Abdul Karim N, Abumrad NA, Acuna-Castroviejo D, Adiele RC, et al (2019) Mitochondrial respiratory states and rates. MitoFit Preprint Arch doi:10.26124/mitofit:190001.v4.|
|Vilks 2018 MiP2018||2018|
|Makarova 2018 MiP2018||2018|
|Makrecka-Kuka 2017 MiP2017||2017|
MitoEAGLE Short-Term Scientific Mission
- Work Plan summary
- I am a third year PhD student at the University of Latvia, Faculty of Biology; and work as research assistant in the Latvian Institute of Organic Synthesis, Laboratory of Pharmaceutical Pharmacology (under supervision of Prof., Dr. Pharm. Maija Dambrova and Dr. Pharm. Edgars Liepinsh).
- The aim of my thesis “Examination of molecular mechanisms of acylcarnitine pathophysiology”, is to characterise the role of long-chain acylcarnitines (LCAC) in development of type 2 diabetes and insulin resistance.
- Obtained results in the present study so far have shown that palmitoylcarnitine (PC) decreases phosphorylation of key kinases of insulin signalling pathway in vitro and in vivo, but further studies are needed to specify the functional outcome of these changes at the cellular level. We have demonstrated that LCAC during ischemia induce cardiac mitochondrial damage and decrease pyruvate and lactate metabolism in isolated mitochondria from heart and brain tissue.
- I am applying for COST STSM to confirm previously obtained results regarding LCAC-induced effects on mitochondrial and extra-mitochondrial glucose metabolism in another laboratory setting using myocyte cell lines. In addition, results obtained in the Muscle Metabolism Research Group (Department of Pharmaceutical Biosciences, School of Pharmacy, University of Oslo) could help to elucidate the role of LCAC in glucose metabolism regulation by providing evidence for functional metabolic changes at the cellular level, such as glycogen production or glucose and fatty acid uptake and oxidation to characterize the switch between energy substrates.
- STSM could provide necessary insight about LCAC regulatory functions in muscles, give necessary data for my PhD thesis, and serve as an excellent opportunity to learn new methods as well as widen transnational collaboration possibilities that could be very important for future research projects.
- In addition, obtained results will be used as a part of publication (in an internationally cited journal), in collaboration with University of Oslo and acknowledgement to the MitoEAGLE (CA15203) network, and used as a pilot data for joint project application next year (EEA Financial Mechanism 2014-2021 Baltic Research Programme call open in 2019).
- Work Plan summary
- 2. Proposed contribution to the scientific objectives of the Action.
- I believe that this STSM will contribute to COST Action MitoEAGLE (CA15203) Working group (WG) 1 and 2 activities because one of the main goals for this mission is to evaluate if observations made in one laboratory regarding LCAC involvement in glucose (pyruvate) metabolism regulation, can be reproduced in another WG 2 laboratory using similar experimental settings, but different assay (respirometry based assay vs radiolabeled substrate based technique) and type of preparation (isolated mitochondria vs cell lines).
- Overall, scientific aim of STSM is to gain valuable insight about regulatory role of muscle LCAC, fatty acid beta oxidation intermediates exclusively produced in mitochondria on glucose (pyruvate/lactate) metabolism regulation. Since one of the goals for this mission is to verify data reproducibility between collaborating laboratories, thus STSM will work on following COST Action MitoEAGLE tasks and objectives:
- • Comparison of instrumental platforms, assays, and type of preparation
- • Optimally harmonisation protocols across research groups
- • Dissemination of updated knowledge and know-how among the partners
- • Forming a unique well-coordinated network of senior researchers and young investigators and establishment a spirit of mentorship and collaboration
- • Initiation application for funding to support international collaborative research projects.
- Moreover, deeper understanding about regulatory and pathophysiological role of acylcarnitines could be used as basis for novel pharmacological compounds which could be used to prevent complications caused by possible mitochondrial disfunction, such as seen during type 2 diabetes and muscle insulin insensitivity, metabolic diseases frequently associated with sedentary lifestyle.
- The results will be used as a part of publication (in an internationally cited journal), in collaboration with University of Oslo and acknowledgement to the MitoEAGLE (CA15203) network and will be used to prepare joint project application (e.g. EEA Financial Mechanisms 2014-2021).
- 3. Techniques - Please detail what techniques or equipment you may learn to use, if applicable.
- Main goal of STSM is to learn selected methods:
- • Myocyte isolation and cultivation from mice and human tissue
- • Scintillation proximity assay (SPA) for real-time measurement of influx or efflux of β- emitting energy substrates non-invasively in 96-well plates
- • CO2 trapping assay- 96-well “plate press” method for capture and quantification of 14CO2 released from cultured cells
- During STSM period 4th -25th of January mentioned methods will be used to evaluate PC induced effects on insulin signalling pathway such as:
- • Glucose and fatty acid uptake in response to PC and insulin treatment
- • Glycogen synthesis
- • Glucose and fatty acid oxidation
- 4. Planning – Please detail the steps you will take to achieve your proposed aim.
- The STSM is planned to take place during the period 4th -25th of January 2019 and is divided in following parts to achieve proposed aims:
- 1st part: arrival, becoming familiar with facilities, protocols, learning techniques (myocyte isolation, SPA and CO2 trapping assays) and design of first experimental trial
- 2nd part: performing experiments to elucidate the role of PC on glucose metabolism in myocytes: SPA and CO2 trapping assay, collection of samples for later analysis at home institution (evaluation of PC effect on insulin signalling in isolated primary cells).
- 3rd part: discussion of obtained data, running second set of experiments, discussion of joint project application and further plan for joint publication.