9 research outputs found
Carbohydrate metabolism.
<p>(A) Heatmap depicting the log basis 2 fold changes in metabolite (rows) concentration determined in liver tissues between matched pairs (columns) of homozygous (G/G, <i>Bcs1l</i><sup>G/G</sup>) and littermate control (A/A, <i>Bcs1l</i><sup>A/A</sup>) mice. Columns (i.e. matched pairs) are reordered by hierarchical clustering (HCA, Ward aggregation method) using the age group (14, 24 and more than 30 days, >30) to label the tree leaves. Metabolites are grouped according to chemical classification. (B) Explicit fold changes with their corresponding 95% confidence intervals for all the carbohydrate intermediates measured in the three age groups. Significance levels: +, q<0.2; **, q<0.01 and ***, q<0.001. DHAP+3-PGA: mixture of dihydroxyacetonephosphate and 3-phosphoglyceraldehyde. (C) Periodic acid-Schiff staining of liver sections showing progressive glycogen depletion from 24 to 34 days old homozygotes.</p
Hypothetic diagram of metabolic changes in hepatocytes in relation to disease progression in <i>Bcs1l</i><sup>G/G</sup> mice.
<p>(A) With decreasing incorporation of Rieske iron sulphur protein (RISP) in complex CIII (CIII), the activity decreases and symptoms appear; at less than 50% incorporation growth failure starts. (B) Metabolomic changes in association with decreased CIII activity and respiratory chain dysfunction are depicted in red. Initial metabolic changes are elevated levels of succinate, aspartate and glutamate. Glycogen depletion leads to lack of glucose and lactate and induces beta-oxidation and protein breakdown resulting in accumulation of fatty acids and acylcarnitines as well as of the biogenic amines putrescine and spermidine. PDH. Pyruvate dehydrogenase complex, CPT1/2: Carnitine palmitoyl transferases 1 and 2. GAP, glyceraldehyde- 3 phosphate, DHAP, Dihydroxy acetone phosphate.</p
Amino acids and biogenic amines.
<p>(A) Log<sub>2</sub> fold changes of amino acids and biogenic amines found significant at false discovery rate <0.1 in the three age groups (14, 24 and >30 days) between matched pairs of homozygous and control mice. Sick homozygotes (indicated as >30 days) have significantly elevated levels of the two metabolite groups, especially the cell signalling amine putrescine.</p
Acylcarnitines and lipid accumulation.
<p>(A) Log<sub>2</sub> fold changes of the acylcarnitines found significant at a false discovery rate <0.1 in the three age groups (14, 24 and >30 days) between matched pairs of homozygous (G/G, <i>Bcs1l</i><sup>G/G</sup>) and control (A/A, <i>Bcs1l</i><sup>A/A</sup>) mice. Metabolites are presented in the increasing order of their molecular weight. Significance levels are defined as: +, q<0.2; *, q<0.05; **, q<0.01 and ***, q<0.001. (B) Oil-Red-O staining showing microvesicular lipid accumulation in hepatocytes of sick animal (34d), only.</p
Respirometry and H<sub>2</sub>O<sub>2</sub> production in mitochondria with RISP depleted CIII (A-C), and signs of oxidative stress in liver tissue (D, E).
<p>(A) Blue Native PAGE and Western blot showing decreased Rieske iron sulphur protein (RISP) incorporation in CIII in mitochondria from two representative <i>Bcs1l</i><sup>G/G</sup> (G/G<sub>1</sub>, G/G<sub>2</sub>) mice compared to wild type (A/A<sub>1</sub>, A/A<sub>2</sub>) aged >30 d. (B) Respiratory chain oxygen consumption under convergent substrate input in CI and CII is impaired in G/G<sub>1</sub>. (C) Similar H<sub>2</sub>O<sub>2</sub> production was detected with Amplex Red in isolated mitochondria of G/G<sub>1</sub> and A/A<sub>1</sub> mice. (D) In liver tissue homogenates of three age group animals, metabolomic analyses showed that log<sub>2</sub> fold changes of the oxidative stress markers were significantly increased in sick animals only (>30 days). (E) Antioxidant defence in liver tissue measured with quantitative PCR in 14 days old and sick (>30 d) homozygotes expressed as relation to β-actin. In sick animals manganese superoxide dismutase (MnSOD) and catalase (Cat) were decreased, whereas glutathione peroxidase 3 (GPx-3) was increased.</p
Impact of propofol on neurotrophins.
<p>Densitometric quantifications of mRNA levels of BDNF and NT-3 in cortex and thalamus of P6 rats, analysed by qRT-PCR. Values represent mean normalised ratios of the densities of BDNF and NT-3 bands compared to the density of the control group (n = 6–7/point+SE). There was an effect of propofol treatment with a decrease of BDNF levels over time, which was significant after 6 hrs in the cortex [F(1,30) = 66.5, p<0.001]. There was also a decrease in NT-3 levels, which was significant in the cortex after 6 hrs [F(1,28) = 12.7, p = 0.004] and after 12 hrs in the thalamus [F(1,24) = 3.5, p = 0.06].</p
Impact of propofol on survival promoting proteins.
<p>Densitometric quantifications of pAKT and pERK1/2 in the cortex and thalamus of P6 rats, analysed by Western blotting. Values represent mean normalised ratios of the densities of pAKT and pERK1/2 bands compared to the density of the control group (n = 6/point+SE). There was an effect of propofol treatment in decrease of pAKT levels over time in the thalamus, which was significant after 12 hrs [F(1,28) = 5.6, p = 0.06]. Post-hoc analysis showed most pronounced decrease after 12 hrs (2-sample t-test). In the cortex there was a significant decrease of pERK1/2 levels over the time, which was significant after 6, 12 and 24 hrs [F(1,29) = 12.7, p = 0.013].</p
Impact of propofol on key proteins involved in apoptotic signalling.
<p>Densitometric quantifications of caspase-3 and AIF in cortex and thalamus of P6 rats as analysed by Western blotting. Values represent mean normalised ratios of the densities of caspase-3 and AIF bands compared to densities of the control group (n = 5–6/point+SE). There was an effect of propofol treatment on caspase-3 levels over time, which was significant after 24 hrs in the cortex [F(1,29) = 3.63, p = 0.06] and after 12 hrs in the thalamus [F(1,28) = 3.1, p = 0.09).</p
Novel object recognition on P30 and P120: At the age of 30 days, both propofol treated animals (t(7) = 7.45, ***q = 4.3×10<sup>−4</sup>) and controls (t(10) = 6.30, ***q = 3.6×10<sup>−4</sup>) spent significantly more time with the novel object indicating their ability to discriminate the novel from the old object.
<p>Propofol (t(7) = −1.44, q = 0.192) as well as control animals (t(10) = −1.92, q = 0.168) failed to do so after a 24 hrs inter-trial interval. At P120 both groups spent a random amount of time with either of the objects after 6 hrs and also after a 24 hrs interval, indicating that they were unable to remember the old object. (n<sub>controls</sub> = 12 animals, n<sub>propofol</sub> = 8 animals).</p