|News and Events||Working Groups||Short-Term Scientific Missions||Management Committee||Members|
COST Action CA15203 (2016-2021): MitoEAGLE
Evolution-Age-Gender-Lifestyle-Environment: mitochondrial fitness mapping
MitoPedia topics: EAGLE
COST WG4: WG4
COST ECI: ECI
|Name||Pereira da Silva Grilo da Silva Filomena, PhD.|
University of Coimbra, PT
Filomena Pereira da Silva Grilo da Silva is a visiting scientist at Oroboros Instruments within the Secondment MtFOIE GRAS from October 2017 to February 2018 and in March 2019.
|Address||Biocant Park, 3060-197|
|BEC 2020.1 doi10.26124bec2020-0001.v1||BEC 2020.1||Gnaiger E et al ― MitoEAGLE Task Group (2020) Mitochondrial physiology. https://doi.org/10.26124/bec:2020-0001.v1|
|Silva 2019 ESCI2019||2019||Silva FSG, Komlodi T, Garcia-Souza LF, Bento G, Doerrier C, Oliveira PJ, Gnaiger E (2019) Can fatty acid oxidation be specifically blocked by the CPT1 inhibitor etomoxir?|
|Silva 2019 MiPschool Coimbra||2019|
|Pereira da Silva Grilo da Silva 2018 MiP2018||2018|
|Silva 2017 MiP2017||2017|
MitoEAGLE Inclusiveness Target Countries - Conference Grant
MitoEAGLE Short-Term Scientific Mission
- Work Plan summary
- 1. The liver mitochondrial dysfunction plays an important role in the progression of non-alcoholic steatohepatitis, therefore improved protocols are required to evaluate mitochondrial fitness in the course of disease. Etomoxir is a well-known compound which irreversibly inhibits the carnitine palmitoyltransferase (CPT-1) in the outer surface of the mitochondrial inner membrane, therefore, blocks the transport of fatty acids into the mitochondrial matrix for further oxidation. However, among others we have shed light on the unspecific effect of etomoxir on the mitochondrial fatty acid oxidation (FAO; F-pathway linked respiration) in permeabilized Huh7 liver cells and mitochondria isolated from mouse liver. In our previous study (data not published) we have observed that the effect of etomoxir is concentration dependent. Low concentration (~40 µM) of etomoxir might inhibit NADH-linked respiration supported by glutamate or pyruvate plus malate. We have seen that high concentration of etomoxir (~200 µM) might block not only the NADH-linked pathway but also the Succinate-linked respiration.
- The aim of the secondment is to adjust the optimal etomoxir concentration which does not exert inhibitory effect towards other respiratory pathways and specifically blocks FAO on mitochondria.
- Work Plan summary
- 2. The goal of this secondment is to provide standardized protocols to study FAO and to deepen our knowledge about the effect of etomoxir on F-pathway linked mitochondrial respiration and get rid of the artefacts created by overdosage of the inhibitor.
- Oroboros laboratory would provide a platform with its instrumental background, with its protocols, its experience and its international team to the objectives and aim of MitoEAGLE.
- 3. High-resolution Respirometry experiments will be performed on isolated mitochondria from mouse liver, heart and brain as well as permeabilized Huh7 liver cell lines. In order to detect FAO, we have used palmitoylcarnitine as source of fatty acid, which needs CPT-1 to enter the mitochondrial matrix and low concentration of malate supporting FAO. We also want to test other sources of fatty acid such as palmitoyl-CoA + carnitine + low amount of malate or palmitate + CoA + carnitine, which might influence the effect of etomoxir on CPT-1.
Visiting scientist in the Oroboros O2k-Laboratory
- March 04 to March 22 2019
- October 01 2016 to February 28 2017