Talk:Fatty acid oxidation
O2k-Network discussion forum: fatty acids used in permeabilized fibre assays (2015-10-20)
Experience with FAO in PFI | Fatty acid used | Mammal and model | Tissue and cell | Coupling states | Pathways | |
---|---|---|---|---|---|---|
Baekkerud 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Octanoylcarnitine + Palmitoylcarnitine | Mouse | F | |||
Bouitbir 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | Heart Skeletal muscle | F | |||
Cannon 2015 Fatty Acid Oxidation O2k-Network Discussion Forum |
| Palmitoylcarnitine | Human Mouse | Skeletal muscle | LEAK OXPHOS ET | F N S NS ROX |
Chavanelle 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Fatty oxidation in permeabilised db/db mice muscle fibres, in Mir05 + Cr +37°C. We found that ADP-stimulated respiration in presence of 0,02 mM of palmitoyl-carnitine (OXPHOS) is very close to the non-ADP-stimulated respiration (LEAK state) when working with these conditions. (Values around 14 to 20 (pmol/s/mg w.t.) for LEAK state and 15 to 24 (pmol/s/mg w.t.) for fatty acid triggered-OXPHOS). These data suggest either a very poor ability for this animal model to use palmitoylcarnitine or a very high LEAK state (or both!)
Protocol
| Palmitoylcarnitine | Mouse | Skeletal muscle | LEAK OXPHOS | F N |
Chicco 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We have used palmitoylcarnitine for most of our studies, but have found it to behave inconsistently in the past, and differently than OctCrn when the two have been compared. So, we have used both, but would be best represented by Palmitoylcarnitine. | Palmitoylcarnitine | F | |||
Chou 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Protocol: final concentration of digitonin in chamber is 10μg/ml cell number in chamber is 2 millions cell/ml, cell type PBMC, Malate (2mM), Palmitoyl-DLcarnitine-HCl (20μM), ADP (2.5mM), pyruvate (5mM), glutamate (10mM), succinate (10mM), rotenone (0.1μM), malonic acid(5mM), myxothiazol (0.5μM), antimycin A (2.5μM), TMPD (0.5mM), azide (100mM) | Palmitoylcarnitine TW | Blood cells Lymphocyte Platelet | OXPHOS ET | F N S NS ROX | |
Ciapaite 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Protocol: I use either palmitoyl-L-carnitine plus malate (25 µM + 2.5 mM) or palmitoyl-CoA + L-carnitine + malate (25 µM + 2 mM + 2.5 mM) as substrates. Respiratory control index is usually around 5-6 for healthy controls. | Palmitoylcarnitine Palmitoyl-CoA + L-carnitine + Malate | Human Rat | Skeletal muscle | LEAK OXPHOS ET | F |
Coen 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We use Palmitylcarnitine as a substrate in assays of permeabilized fiber bundle respiration. The substrate works well for us and in combination with Pal, Mal, Glut, and Succ we see higher levels of state 3 respiration than without it. | Palmitoylcarnitine | OXPHOS | F N NS | ||
Darveau 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | I have conducted one study examining bumblebee flight muscle permeabilized fibers and used. | Palmitoylcarnitine | Hexapods | Skeletal muscle | F | |
Egelandsdal 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We have examined for how long time post mortem the fatty acid oxidation prevails in beef muscles. | Octanoylcarnitine | Bovines | Skeletal muscle | F | |
Essop 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | F | ||||
Fontes-Oliveira 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine + Malate | F | ||||
Goodpaster 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Excellent. Lower overall O2 consumption compared to carbohydrate-supported respiration | Palmitoylcarnitine | F | |||
Gorr 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | What´s wrong with using simple C16 saturated or C18 monounsaturated fatty acids like palmitate or oleate, respectively, for analysing beta-oxidation rates?? | F | ||||
Gottschalk 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | F | ||||
Hansson Fatty Acid Oxidation O2k-Network Discussion Forum | We have only performed limited pilot studies with fatty acids in permeabilized heart fibers. In general the respiration by both octanoylcarnitine and palmitylcarnitine have been quite low. We have not optimized the settings so we do not have much valuable experience to share. | Octanoylcarnitine Palmitoylcarnitine | Heart | F | ||
Hickey 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We use Palmitoyl Carnitine, I recall we made it up with fatty acid free BSA at 5:1 (PC:BSA). Worked well with fish and rat heart mitochondria, and permeabilised fibres. Protocol: Palmitoylcarnitine we made it with fatty acid free BSA at 5:1 (PC:BSA) | Palmitoylcarnitine | Rat Fishes | Heart | F | |
Jelenik 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We standardly measure FA-linked respiration in the muscle - we used Octanoyl-C as well as Palmitoyl-C, however, the latter one gives poor response. Thus we use octanoyl. | Octanoylcarnitine | F | |||
Kane 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | F | ||||
Lemieux 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Heart and skeletal muscle. Permeabilized animals (Dugesia tigrina worm) Other substrates: Acetylcarnitine + Malate, Palmitoyl-CoA+Malate+Carnitine | Octanoylcarnitine + Malate Palmitoylcarnitine + Malate | Heart Skeletal muscle | F | ||
Lesnefsky 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We use the method pioneered by Dr. Chuck Hoppel in Cleveland. We use palmitoylcarnitine. To assess role of CPT-1; one can use palmitoyl-CoA + carnitine | Palmitoylcarnitine Palmitoyl-CoA + Carnitine | F | |||
Ljubkovic 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | I am sending you a graph with some mitochondrial respiration measurements obtained from the left ventricular human tissue of patients undergoing CABG surgery. As can be seen from the graph, we used FFA substrate to evoke the State 2 and State 3 respiration, and then followed with other substrates and an uncoupler. FFA substrate was palmitoyl-carnitine (40 µM concentration). We performed quite a few of experiments using palmitoyl-carnitine, also in the range of concentrations from 1 – 40 µM. We usually see a gradual increase in State 3 respiration with concentration increments (1, 2.5, 5, 10, 15, 40 µM). However, the respiration is always lower then when pyruvate is used (although in much higher concentration, from 2 mM up). All of the observations come from the human left ventricular tissue, that we almost exclusively use in past couple of years for our experiments on mitochondria (we also measured activity of some enzymes and protein expression). | Palmitoylcarnitine | Human | Heart | OXPHOS ET | F N NS |
Madlala 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We have assessed beta oxidation in permeabilised skeletal muscle fibres. We initially used palmitoylcarnitine in our analysis but we realised that using this substrate for assessing beta oxidation in skeletal muscle of rat does not work (no response in respiration). This is also documented in literature; absence of LCAD in rat skeletal muscle so we decided to use octanoylcarnitine instead and it works well. Afterwards the muscle is responsive to ADP and uncoupler titrations. However, we use this in combination with malate. | Octanoylcarnitine + Malate Palmitoylcarnitine | Rat | Skeletal muscle | OXPHOS | F |
Makrecka 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We have examined fatty acid oxidation in permeabilized muscle fibres and cardiac fibres and in isolated cardiac mitochondria of rats and mice. Since the area of our research is the regulation of energy metabolism, the determination of fatty acid oxidation is one of the typical measurements. Which substrate we use, is dependent on the aim (include CPT I (rate limiting step) or not). In addition, presence of BSA in media and used carnitine concentration are always under discussions (physiological or not physiological). | Palmitoylcarnitine + Malate | Mouse Rat | Heart Skeletal muscle | F | |
McCurdy 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We have had good success using the Octanoylcarnitine at the recommended dose of 0.2mM in SUIT protocols with PMG and with just GM. Respiration increases with addition of ADP and subsequent substrate additions. We have also used palmitoylcarnitine at a conc of 0.025 -0.05mM with mixed success. Often there is no or little stable increase in ADP stimulated respiration and requires addition of pyr and/or Glut. We primarily test nonhuman primate (japanses macaque) permeabilized muscle fibers. | Octanoylcarnitine + Palmitoylcarnitine | Other mammals | Skeletal muscle | OXPHOS | F N |
Miller 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Octanoylcarnitine | F | ||||
Moellering 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We use submaximal concentrations 40µM of palmitoylcarnitine with our permeabilized muscle fiber preps. | Palmitoylcarnitine | Skeletal muscle | F | ||
Neviere 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | Human Mouse | F | |||
Ojuka 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We have used Octanoylcarnitine + malate as fatty acid substrates (at the concentration stipulated in BIOBLAST) with much success. However, Palmitylcarnitine + malate was not responsive to ADP or CCCP. | Octanoylcarnitine + Malate | OXPHOS ET | F | ||
Ost 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Right now we´re using octanoyl carnitine (OC) when examining mouse skeletal muscle permeabilized fiber respiration. We ususally start with Malate (2mM) and OC (0.2mM), then add ADP (5mM) in order to assess fatty acid oxidative capacity. For Soleus muscle fibers we got an average respiration of 60 pmol/(s*mg fiber ww). | Octanoylcarnitine Malate ADP | Mouse | Skeletal muscle | F | |
Pasdois 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Protocol: Couple palmitoylcarnitine (10µM) + malate (1mM) on isolated mitochondria and permeabilized fibers. In such case the buffer is always supplemented with 2mg/ml of BSA. | Palmitoylcarnitine + Malate | F | |||
Ramsay 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | I recommend that everyone uses palmitoyl-Cn (with BSA of course) for the most physiological measurement. I actually used decanoyl-CoA when I wanted a more soluble substrate. | Palmitoyl-Cn Decanoyl-CoA | F | |||
Roden 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Great experience | Octanoylcarnitine + Palmitoylcarnitine | Human Mouse | Skeletal muscle | F | |
Salin 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Worked a bit with palmitoylcarnitine on homogenate of white muscle of fish. Do not need to be permeabilized chemically as mechanical permeabilization allow to obtain permeabilized cell (ADP-responsiveness the same). The respiration rate was much lower than the pyruvate / malate / succinate substrate, so I stopped to use the lipid substrates. | Palmitoylcarnitine | Fishes | Skeletal muscle | F | |
Schauer 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We are successfully measuring FA oxidation in permeabilized fibers. We have switched to Octanoylcarnitine (A) because we were seeing evidence of toxicity with palmitoylcarnitine. This evidence was that we were seeing final uncoupled respiration to be lower than state 3 respiration in samples with PC. This issue resolved when we switched to OC. | Octanoylcarnitine | OXPHOS ET | F | ||
Senyilmaz 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Inverted Drosophila larvae which are permeabilized by digitonin. Their body walls are more or less made of muscles. | Palmitoyl-CoA + Carnitine | Drosophila | Skeletal muscle | F | |
Smith 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | I have extensively tested FAO in PMFBs using the Palmitoylcarnitine, Palmitoyl-CoA, and L-Carnitine. Protocol: For maximal respiration we make sequential additions of 18 µM PC, 20 µM PCoA, and 5 mM Carnitine. Addition of ADP induces a peak rate of O2 consumption that is not increased with FCCP addition. | Palmitoylcarnitine Palmitoyl-CoA and L-Carnitine | OXPHOS ET | F | ||
Sparks 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We routinely (up to 4 human and mouse studies) measure respiration supported by glycolytic and fatty acid substrates using SUIT protocols in permeabilized muscle fibers. We always use palmitoylcarnitine. In the past, in Dr. Schrauwen’s lab, we used octanoate to titrate for fatty acid induced uncoupling. | Palmitoylcarnitine Octanoate | Human Mouse | Skeletal muscle | F | |
Van Schaardenburgh 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | In my studies on patients with intermittent claudication we used Octanoylcarnitine in the SUIT protocol described by Pesta and you. | Octanoylcarnitine | F | |||
Veksler 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We studied fatty acid oxidation in permeabilised cardiac and skeletal muscle fibers for 15 years. We use palmitoylcarnitine, palmitoyl-CoA or octanoate | Palmitoylcarnitine Palmitoyl-CoA Octanoate | Heart Skeletal muscle | F | ||
Votion 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Very limited (only preliminary tests on mice) but satisfying. I have planned new experiments on equine muscle tissue in 2016. | Octanoylcarnitine Palmitoylcarnitine Octanoate Acetylcarnitine Palmitoyl CoA (+ Carnitine) | Mouse | F | ||
West 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Octanoylcarnitine | F | ||||
Wohlwend 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Fatty acid oxidation in permeabilized muscle fibers has worked well for us in both human and rat studies. Typically we use Octanoylcarnitine and Malate for skeletal muscle and a combination of Octanoylcarnitine plus Palmitylcarnitine and Malate for human heart tissue. These protocols have given us a good insight into Beta oxidation. | Octanoylcarnitine + Malate Octanoylcarnitine + Palmitoylcarnitine + Malate (for heart) | Human Rat | Skeletal muscle | F | |
Wright 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We do include octanoylcarnitine in the initial titration of substrates for our SUIT protocols typically, but do not measure its individual contribution to OXPHOS. | Octanoylcarnitine | F | |||
Wuest 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Compared with just glucose oxidation, the max OXPHOS/ET capacity was slightly (but significantly) lower with PC & Oct. Protocol: PC, M, ADP, OCT, G, P, SUC, (FCCP, rat), ROT, AA. The concentrations were the same as in the blue book. | Palmitoylcarnitine + Octanoylcarnitine | Rat | Heart | OXPHOS ET | F N S NS ROX |
O2k-Network discussion forum: fatty acids all data (2015-10-20)
Experience with FAO in PFI | Fatty acid used | Mammal and model | Tissue and cell | Coupling states | Pathways | |
---|---|---|---|---|---|---|
Ali 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | F | ||||
Baekkerud 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Octanoylcarnitine + Palmitoylcarnitine | Mouse | F | |||
Bouitbir 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | Heart Skeletal muscle | F | |||
Cabrera-Fuentes 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | F | ||||
Cannon 2015 Fatty Acid Oxidation O2k-Network Discussion Forum |
| Palmitoylcarnitine | Human Mouse | Skeletal muscle | LEAK OXPHOS ET | F N S NS ROX |
Chavanelle 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Fatty oxidation in permeabilised db/db mice muscle fibres, in Mir05 + Cr +37°C. We found that ADP-stimulated respiration in presence of 0,02 mM of palmitoyl-carnitine (OXPHOS) is very close to the non-ADP-stimulated respiration (LEAK state) when working with these conditions. (Values around 14 to 20 (pmol/s/mg w.t.) for LEAK state and 15 to 24 (pmol/s/mg w.t.) for fatty acid triggered-OXPHOS). These data suggest either a very poor ability for this animal model to use palmitoylcarnitine or a very high LEAK state (or both!)
Protocol
| Palmitoylcarnitine | Mouse | Skeletal muscle | LEAK OXPHOS | F N |
Chicco 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We have used palmitoylcarnitine for most of our studies, but have found it to behave inconsistently in the past, and differently than OctCrn when the two have been compared. So, we have used both, but would be best represented by Palmitoylcarnitine. | Palmitoylcarnitine | F | |||
Chou 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Protocol: final concentration of digitonin in chamber is 10μg/ml cell number in chamber is 2 millions cell/ml, cell type PBMC, Malate (2mM), Palmitoyl-DLcarnitine-HCl (20μM), ADP (2.5mM), pyruvate (5mM), glutamate (10mM), succinate (10mM), rotenone (0.1μM), malonic acid(5mM), myxothiazol (0.5μM), antimycin A (2.5μM), TMPD (0.5mM), azide (100mM) | Palmitoylcarnitine TW | Blood cells Lymphocyte Platelet | OXPHOS ET | F N S NS ROX | |
Ciapaite 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Protocol: I use either palmitoyl-L-carnitine plus malate (25 µM + 2.5 mM) or palmitoyl-CoA + L-carnitine + malate (25 µM + 2 mM + 2.5 mM) as substrates. Respiratory control index is usually around 5-6 for healthy controls. | Palmitoylcarnitine Palmitoyl-CoA + L-carnitine + Malate | Human Rat | Skeletal muscle | LEAK OXPHOS ET | F |
Coen 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We use Palmitylcarnitine as a substrate in assays of permeabilized fiber bundle respiration. The substrate works well for us and in combination with Pal, Mal, Glut, and Succ we see higher levels of state 3 respiration than without it. | Palmitoylcarnitine | OXPHOS | F N NS | ||
Cortassa 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoyl CoA (mitochondria) and Palmitate (conjugated with BSA) in cardiomyocytes. | Palmitoyl CoA Palmitate | Skeletal muscle | F | ||
Darveau 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | I have conducted one study examining bumblebee flight muscle permeabilized fibers and used. | Palmitoylcarnitine | Hexapods | Skeletal muscle | F | |
Egelandsdal 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We have examined for how long time post mortem the fatty acid oxidation prevails in beef muscles. | Octanoylcarnitine | Bovines | Skeletal muscle | F | |
Essop 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | F | ||||
Filippi 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Octanoylcarnitine Oleate | Liver | F | |||
Fontes-Oliveira 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine + Malate | F | ||||
Goetzman 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | Liver | F | |||
Goodpaster 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Excellent. Lower overall O2 consumption compared to carbohydrate-supported respiration | Palmitoylcarnitine | F | |||
Gorr 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | What´s wrong with using simple C16 saturated or C18 monounsaturated fatty acids like palmitate or oleate, respectively, for analysing beta-oxidation rates?? | F | ||||
Gottlieb 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We used palmitate added to culture media overnight in differentiated C2C12 myotubes. | Palmitate | Mouse | Skeletal muscle | F | |
Gottschalk 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | F | ||||
Hagland 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitic acid | F | ||||
Hansson Fatty Acid Oxidation O2k-Network Discussion Forum | We have only performed limited pilot studies with fatty acids in permeabilized heart fibers. In general the respiration by both octanoylcarnitine and palmitylcarnitine have been quite low. We have not optimized the settings so we do not have much valuable experience to share. | Octanoylcarnitine Palmitoylcarnitine | Heart | F | ||
Hickey 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We use Palmitoyl Carnitine, I recall we made it up with fatty acid free BSA at 5:1 (PC:BSA). Worked well with fish and rat heart mitochondria, and permeabilised fibres. Protocol: Palmitoylcarnitine we made it with fatty acid free BSA at 5:1 (PC:BSA) | Palmitoylcarnitine | Rat Fishes | Heart | F | |
Jelenik 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We standardly measure FA-linked respiration in the muscle - we used Octanoyl-C as well as Palmitoyl-C, however, the latter one gives poor response. Thus we use octanoyl. | Octanoylcarnitine | F | |||
Kaambre 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | Human | Genital | F | ||
Kane 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | F | ||||
Kollberg 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | F | ||||
Krishnan 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | Liver | F | |||
Leloup 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitate | Nervous system | ||||
Lemieux 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Heart and skeletal muscle. Permeabilized animals (Dugesia tigrina worm) Other substrates: Acetylcarnitine + Malate, Palmitoyl-CoA+Malate+Carnitine | Octanoylcarnitine + Malate Palmitoylcarnitine + Malate | Heart Skeletal muscle | F | ||
Lesnefsky 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We use the method pioneered by Dr. Chuck Hoppel in Cleveland. We use palmitoylcarnitine. To assess role of CPT-1; one can use palmitoyl-CoA + carnitine | Palmitoylcarnitine Palmitoyl-CoA + Carnitine | F | |||
Ljubkovic 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | I am sending you a graph with some mitochondrial respiration measurements obtained from the left ventricular human tissue of patients undergoing CABG surgery. As can be seen from the graph, we used FFA substrate to evoke the State 2 and State 3 respiration, and then followed with other substrates and an uncoupler. FFA substrate was palmitoyl-carnitine (40 µM concentration). We performed quite a few of experiments using palmitoyl-carnitine, also in the range of concentrations from 1 – 40 µM. We usually see a gradual increase in State 3 respiration with concentration increments (1, 2.5, 5, 10, 15, 40 µM). However, the respiration is always lower then when pyruvate is used (although in much higher concentration, from 2 mM up). All of the observations come from the human left ventricular tissue, that we almost exclusively use in past couple of years for our experiments on mitochondria (we also measured activity of some enzymes and protein expression). | Palmitoylcarnitine | Human | Heart | OXPHOS ET | F N NS |
Lund 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Carnitine + Palmitoyl CoA | F | ||||
Madlala 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We have assessed beta oxidation in permeabilised skeletal muscle fibres. We initially used palmitoylcarnitine in our analysis but we realised that using this substrate for assessing beta oxidation in skeletal muscle of rat does not work (no response in respiration). This is also documented in literature; absence of LCAD in rat skeletal muscle so we decided to use octanoylcarnitine instead and it works well. Afterwards the muscle is responsive to ADP and uncoupler titrations. However, we use this in combination with malate. | Octanoylcarnitine + Malate Palmitoylcarnitine | Rat | Skeletal muscle | OXPHOS | F |
Mahdaviani 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | Fat | F | |||
Makrecka 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We have examined fatty acid oxidation in permeabilized muscle fibres and cardiac fibres and in isolated cardiac mitochondria of rats and mice. Since the area of our research is the regulation of energy metabolism, the determination of fatty acid oxidation is one of the typical measurements. Which substrate we use, is dependent on the aim (include CPT I (rate limiting step) or not). In addition, presence of BSA in media and used carnitine concentration are always under discussions (physiological or not physiological). | Palmitoylcarnitine + Malate | Mouse Rat | Heart Skeletal muscle | F | |
McCurdy 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We have had good success using the Octanoylcarnitine at the recommended dose of 0.2mM in SUIT protocols with PMG and with just GM. Respiration increases with addition of ADP and subsequent substrate additions. We have also used palmitoylcarnitine at a conc of 0.025 -0.05mM with mixed success. Often there is no or little stable increase in ADP stimulated respiration and requires addition of pyr and/or Glut. We primarily test nonhuman primate (japanses macaque) permeabilized muscle fibers. | Octanoylcarnitine + Palmitoylcarnitine | Other mammals | Skeletal muscle | OXPHOS | F N |
Miller 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Octanoylcarnitine | F | ||||
Moellering 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We use submaximal concentrations 40µM of palmitoylcarnitine with our permeabilized muscle fiber preps. | Palmitoylcarnitine | Skeletal muscle | F | ||
Neviere 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | Human Mouse | F | |||
Ojuka 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We have used Octanoylcarnitine + malate as fatty acid substrates (at the concentration stipulated in BIOBLAST) with much success. However, Palmitylcarnitine + malate was not responsive to ADP or CCCP. | Octanoylcarnitine + Malate | OXPHOS ET | F | ||
Ost 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Right now we´re using octanoyl carnitine (OC) when examining mouse skeletal muscle permeabilized fiber respiration. We ususally start with Malate (2mM) and OC (0.2mM), then add ADP (5mM) in order to assess fatty acid oxidative capacity. For Soleus muscle fibers we got an average respiration of 60 pmol/(s*mg fiber ww). | Octanoylcarnitine Malate ADP | Mouse | Skeletal muscle | F | |
Palm 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine | F | ||||
Pasdois 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Protocol: Couple palmitoylcarnitine (10µM) + malate (1mM) on isolated mitochondria and permeabilized fibers. In such case the buffer is always supplemented with 2mg/ml of BSA. | Palmitoylcarnitine + Malate | F | |||
Ramsay 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | I recommend that everyone uses palmitoyl-Cn (with BSA of course) for the most physiological measurement. I actually used decanoyl-CoA when I wanted a more soluble substrate. | Palmitoyl-Cn Decanoyl-CoA | F | |||
Robidoux 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Protocol: Palmitate, Stearate, Oleate and Linoleate in intact cells. We use various BSA-fatty acid combinations that result in free fatty acid levels that are in the 2 to 12 nM range. | Palmitate Stearate Oleate Linoleate | Fat | F | ||
Roden 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Great experience | Octanoylcarnitine + Palmitoylcarnitine | Human Mouse | Skeletal muscle | F | |
Salin 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Worked a bit with palmitoylcarnitine on homogenate of white muscle of fish. Do not need to be permeabilized chemically as mechanical permeabilization allow to obtain permeabilized cell (ADP-responsiveness the same). The respiration rate was much lower than the pyruvate / malate / succinate substrate, so I stopped to use the lipid substrates. | Palmitoylcarnitine | Fishes | Skeletal muscle | F | |
Schauer 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We are successfully measuring FA oxidation in permeabilized fibers. We have switched to Octanoylcarnitine (A) because we were seeing evidence of toxicity with palmitoylcarnitine. This evidence was that we were seeing final uncoupled respiration to be lower than state 3 respiration in samples with PC. This issue resolved when we switched to OC. | Octanoylcarnitine | OXPHOS ET | F | ||
Schoettl 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Beta oxidation in adipocytes (white and brown), HEK293 cells, isolated mitochondria from liver, adipocytes, brain, etc. | Palmitoylcarnitine | Nervous system Liver Fat HEK | F | ||
Senyilmaz 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Inverted Drosophila larvae which are permeabilized by digitonin. Their body walls are more or less made of muscles. | Palmitoyl-CoA + Carnitine | Drosophila | Skeletal muscle | F | |
Smith 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | I have extensively tested FAO in PMFBs using the Palmitoylcarnitine, Palmitoyl-CoA, and L-Carnitine. Protocol: For maximal respiration we make sequential additions of 18 µM PC, 20 µM PCoA, and 5 mM Carnitine. Addition of ADP induces a peak rate of O2 consumption that is not increased with FCCP addition. | Palmitoylcarnitine Palmitoyl-CoA and L-Carnitine | OXPHOS ET | F | ||
Sparks 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We routinely (up to 4 human and mouse studies) measure respiration supported by glycolytic and fatty acid substrates using SUIT protocols in permeabilized muscle fibers. We always use palmitoylcarnitine. In the past, in Dr. Schrauwen’s lab, we used octanoate to titrate for fatty acid induced uncoupling. | Palmitoylcarnitine Octanoate | Human Mouse | Skeletal muscle | F | |
Trifunovic 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Palmitoylcarnitine works better than Octanoylcarnitine. | Octanoylcarnitine Palmitoylcarnitine | Heart | F | ||
Van Schaardenburgh 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | In my studies on patients with intermittent claudication we used Octanoylcarnitine in the SUIT protocol described by Pesta and you. | Octanoylcarnitine | F | |||
Veksler 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We studied fatty acid oxidation in permeabilised cardiac and skeletal muscle fibers for 15 years. We use palmitoylcarnitine, palmitoyl-CoA or octanoate | Palmitoylcarnitine Palmitoyl-CoA Octanoate | Heart Skeletal muscle | F | ||
Villena 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | C14-labelled palmitic acid | Fat | F | |||
Votion 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Very limited (only preliminary tests on mice) but satisfying. I have planned new experiments on equine muscle tissue in 2016. | Octanoylcarnitine Palmitoylcarnitine Octanoate Acetylcarnitine Palmitoyl CoA (+ Carnitine) | Mouse | F | ||
Weber 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Octanoylcarnitine | Human Mouse | Genital | F | ||
West 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Octanoylcarnitine | F | ||||
Wohlwend 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Fatty acid oxidation in permeabilized muscle fibers has worked well for us in both human and rat studies. Typically we use Octanoylcarnitine and Malate for skeletal muscle and a combination of Octanoylcarnitine plus Palmitylcarnitine and Malate for human heart tissue. These protocols have given us a good insight into Beta oxidation. | Octanoylcarnitine + Malate Octanoylcarnitine + Palmitoylcarnitine + Malate (for heart) | Human Rat | Skeletal muscle | F | |
Wright 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | We do include octanoylcarnitine in the initial titration of substrates for our SUIT protocols typically, but do not measure its individual contribution to OXPHOS. | Octanoylcarnitine | F | |||
Wuest 2015 Fatty Acid Oxidation O2k-Network Discussion Forum | Compared with just glucose oxidation, the max OXPHOS/ET capacity was slightly (but significantly) lower with PC & Oct. Protocol: PC, M, ADP, OCT, G, P, SUC, (FCCP, rat), ROT, AA. The concentrations were the same as in the blue book. | Palmitoylcarnitine + Octanoylcarnitine | Rat | Heart | OXPHOS ET | F N S NS ROX |
FA and cell toxicity
I am working at the University of Colorado with Drs Paul MacLean and Matthew Jackman who acquired two OROBOROS this year. Following our collaborators advice (Dr Donal O'Gorman at Dublin City Uni), we are following your protocol to measure fat oxidation. Contrary to results in the literature and from Dr O'Gorman's lab, as soon as we add L-palmitoylcarnitine, the cells die. 2 uM only of L-PC are sufficient to reach a respiration of zero. Adding any other substrates does not allow an increase in O2 consumption. Our local collaborator Dr Jane Reush is encountering the same issue and therefore only use octanoyl carnitine. Octanoyl carnitine does not alter mitochondria/cell. Following advices from Dr Neufer (North Carolina), we added some carnitine. This did not allow maintaining the integrity of the cell. (1)
Because we are in altitude (1600m), we need to use a piO2 of 200-220 (instead of 160) in basal and of 350 (instead of 300) in maximal. We are wondering if this higher piO2 could promote the generation of ROS species or oxidative stress that would kill the mitochondria/cell when adding L-PC? (2)
- Audrey (2015-05-14)
- O2k-Network: US CO Aurora Jackman MR
- Audrey Bergouignan, PhD
- Anschutz Health and Wellness Center Division of Endocrinology, Metabolism and Diabetes
- University of Colorado
- E [email protected]
- http://www.anschutzwellness.com/
Response
- »Fatty acid oxidation#FAO and HRR
- When using permeabilized muscle fibers, we have to increase oxygen levels to nonphysiological hyperoxia to avoid oxygen diffusion limitation to the fibers. Although high oxygen levels stimulate ROS production, mitochondria remain functional during times of measurement up to 3 hours. (Gnaiger E, 2015-05-15)
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