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Decreased myocardial injury and improved contractility after administration of a peptide derived against the alpha-interacting domain of the L-type calcium channel.

Viola HM, Jordan MC, Roos KP, Hool LC - J Am Heart Assoc (2014)

Bottom Line: Activation of the channel also alters mitochondrial function.In search of the mechanism for the effect, we found that intracellular calcium ([Ca(2+)]i, Fura-2), superoxide production (dihydroethidium fluorescence), mitochondrial membrane potential (Ψm, JC-1 fluorescence), reduced nicotinamide adenine dinucleotide production, and flavoprotein oxidation (autofluorescence) are decreased after application of AID-TAT peptide.Application of AID-TAT peptide significantly decreased infarct size and supported contractility up to 12 weeks postcoronary artery occlusion as a result of a decrease in metabolic demand during reperfusion.

View Article: PubMed Central - PubMed

Affiliation: School of Anatomy, Physiology and Human Biology, The University of Western Australia, Crawley, WA, Australia (H.M.V., L.C.H.).

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Effect of AID‐TAT peptide on reduction of NAD+ to NADH in vitro. A, Representative traces of NADH autofluorescence recorded in a myocyte before and after 5 minutes of exposure to 40 μmol/L of H2O2 followed by 5 minutes of exposure to 10 U/mL of catalase and a myocyte before and after 5 minutes of exposure to 0 μmol/L of H2O2 followed by 5 minutes of exposure to 10 U/mL of catalase. Vertical arrows indicate when drugs were added. NADH fluorescence decreased after addition of 4 mmol/L of sodium cyanide (NaCN), consistent with depletion of NADH after collapse of Ψm. B, Mean±SEM of changes in NADH autofluorescence for all myocytes exposed to 40 μmol/L of H2O2, 10 U/mL of catalase, and 10 μmol/L of nisoldipine (Nisol) as indicated. C, Representative traces of NADH autofluorescence recorded in myocytes before and after 5 minutes of exposure to 40 μmol/L of H2O2 followed by 5 minutes of exposure to 10 U/mL of catalase in the presence of either 1 or 10 μmol/L of AID(S)‐TAT or AID‐TAT peptide as indicated. Vertical arrows indicate when drugs were added. NADH fluorescence decreased after addition of 4 mmol/L of NaCN, consistent with depletion of NADH after collapse of Ψm. D, Mean±SEM of changes in NADH autofluorescence for all myocytes exposed to 40 μmol/L of H2O2, 10 U/mL of catalase, and 1 or 10 μmol/L of AID(S)‐TAT or AID‐TAT peptide as indicated. Statistical significance was determined using the Kruskal‐Wallis test followed by Dunn's multiple comparison test. AID indicates alpha‐interacting domain; NAD+, nicotinamide adenine dinucleotide; NADH, reduced nicotinamide adenine dinucleotide; TAT, transactivator of transcription.
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fig05: Effect of AID‐TAT peptide on reduction of NAD+ to NADH in vitro. A, Representative traces of NADH autofluorescence recorded in a myocyte before and after 5 minutes of exposure to 40 μmol/L of H2O2 followed by 5 minutes of exposure to 10 U/mL of catalase and a myocyte before and after 5 minutes of exposure to 0 μmol/L of H2O2 followed by 5 minutes of exposure to 10 U/mL of catalase. Vertical arrows indicate when drugs were added. NADH fluorescence decreased after addition of 4 mmol/L of sodium cyanide (NaCN), consistent with depletion of NADH after collapse of Ψm. B, Mean±SEM of changes in NADH autofluorescence for all myocytes exposed to 40 μmol/L of H2O2, 10 U/mL of catalase, and 10 μmol/L of nisoldipine (Nisol) as indicated. C, Representative traces of NADH autofluorescence recorded in myocytes before and after 5 minutes of exposure to 40 μmol/L of H2O2 followed by 5 minutes of exposure to 10 U/mL of catalase in the presence of either 1 or 10 μmol/L of AID(S)‐TAT or AID‐TAT peptide as indicated. Vertical arrows indicate when drugs were added. NADH fluorescence decreased after addition of 4 mmol/L of NaCN, consistent with depletion of NADH after collapse of Ψm. D, Mean±SEM of changes in NADH autofluorescence for all myocytes exposed to 40 μmol/L of H2O2, 10 U/mL of catalase, and 1 or 10 μmol/L of AID(S)‐TAT or AID‐TAT peptide as indicated. Statistical significance was determined using the Kruskal‐Wallis test followed by Dunn's multiple comparison test. AID indicates alpha‐interacting domain; NAD+, nicotinamide adenine dinucleotide; NADH, reduced nicotinamide adenine dinucleotide; TAT, transactivator of transcription.

Mentions: We assessed changes in mitochondrial NADH as autofluorescence in myocytes. Consistent with previous results,19 addition of H2O2 significantly increased NADH signal that could be attenuated with nisoldipine (Figure 5A and 5B). Collapse of the signal with addition of NaCN indicated that the signal was mitochondrial in origin. Application of 1 and 10 μmol/L of AID‐TAT peptide attenuated the increase in NADH associated with application of H2O2 (Figure 5C and 5D). This suggested that AID‐TAT peptide was altering NADH as a result of effects on MMP or oxygen consumption.


Decreased myocardial injury and improved contractility after administration of a peptide derived against the alpha-interacting domain of the L-type calcium channel.

Viola HM, Jordan MC, Roos KP, Hool LC - J Am Heart Assoc (2014)

Effect of AID‐TAT peptide on reduction of NAD+ to NADH in vitro. A, Representative traces of NADH autofluorescence recorded in a myocyte before and after 5 minutes of exposure to 40 μmol/L of H2O2 followed by 5 minutes of exposure to 10 U/mL of catalase and a myocyte before and after 5 minutes of exposure to 0 μmol/L of H2O2 followed by 5 minutes of exposure to 10 U/mL of catalase. Vertical arrows indicate when drugs were added. NADH fluorescence decreased after addition of 4 mmol/L of sodium cyanide (NaCN), consistent with depletion of NADH after collapse of Ψm. B, Mean±SEM of changes in NADH autofluorescence for all myocytes exposed to 40 μmol/L of H2O2, 10 U/mL of catalase, and 10 μmol/L of nisoldipine (Nisol) as indicated. C, Representative traces of NADH autofluorescence recorded in myocytes before and after 5 minutes of exposure to 40 μmol/L of H2O2 followed by 5 minutes of exposure to 10 U/mL of catalase in the presence of either 1 or 10 μmol/L of AID(S)‐TAT or AID‐TAT peptide as indicated. Vertical arrows indicate when drugs were added. NADH fluorescence decreased after addition of 4 mmol/L of NaCN, consistent with depletion of NADH after collapse of Ψm. D, Mean±SEM of changes in NADH autofluorescence for all myocytes exposed to 40 μmol/L of H2O2, 10 U/mL of catalase, and 1 or 10 μmol/L of AID(S)‐TAT or AID‐TAT peptide as indicated. Statistical significance was determined using the Kruskal‐Wallis test followed by Dunn's multiple comparison test. AID indicates alpha‐interacting domain; NAD+, nicotinamide adenine dinucleotide; NADH, reduced nicotinamide adenine dinucleotide; TAT, transactivator of transcription.
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fig05: Effect of AID‐TAT peptide on reduction of NAD+ to NADH in vitro. A, Representative traces of NADH autofluorescence recorded in a myocyte before and after 5 minutes of exposure to 40 μmol/L of H2O2 followed by 5 minutes of exposure to 10 U/mL of catalase and a myocyte before and after 5 minutes of exposure to 0 μmol/L of H2O2 followed by 5 minutes of exposure to 10 U/mL of catalase. Vertical arrows indicate when drugs were added. NADH fluorescence decreased after addition of 4 mmol/L of sodium cyanide (NaCN), consistent with depletion of NADH after collapse of Ψm. B, Mean±SEM of changes in NADH autofluorescence for all myocytes exposed to 40 μmol/L of H2O2, 10 U/mL of catalase, and 10 μmol/L of nisoldipine (Nisol) as indicated. C, Representative traces of NADH autofluorescence recorded in myocytes before and after 5 minutes of exposure to 40 μmol/L of H2O2 followed by 5 minutes of exposure to 10 U/mL of catalase in the presence of either 1 or 10 μmol/L of AID(S)‐TAT or AID‐TAT peptide as indicated. Vertical arrows indicate when drugs were added. NADH fluorescence decreased after addition of 4 mmol/L of NaCN, consistent with depletion of NADH after collapse of Ψm. D, Mean±SEM of changes in NADH autofluorescence for all myocytes exposed to 40 μmol/L of H2O2, 10 U/mL of catalase, and 1 or 10 μmol/L of AID(S)‐TAT or AID‐TAT peptide as indicated. Statistical significance was determined using the Kruskal‐Wallis test followed by Dunn's multiple comparison test. AID indicates alpha‐interacting domain; NAD+, nicotinamide adenine dinucleotide; NADH, reduced nicotinamide adenine dinucleotide; TAT, transactivator of transcription.
Mentions: We assessed changes in mitochondrial NADH as autofluorescence in myocytes. Consistent with previous results,19 addition of H2O2 significantly increased NADH signal that could be attenuated with nisoldipine (Figure 5A and 5B). Collapse of the signal with addition of NaCN indicated that the signal was mitochondrial in origin. Application of 1 and 10 μmol/L of AID‐TAT peptide attenuated the increase in NADH associated with application of H2O2 (Figure 5C and 5D). This suggested that AID‐TAT peptide was altering NADH as a result of effects on MMP or oxygen consumption.

Bottom Line: Activation of the channel also alters mitochondrial function.In search of the mechanism for the effect, we found that intracellular calcium ([Ca(2+)]i, Fura-2), superoxide production (dihydroethidium fluorescence), mitochondrial membrane potential (Ψm, JC-1 fluorescence), reduced nicotinamide adenine dinucleotide production, and flavoprotein oxidation (autofluorescence) are decreased after application of AID-TAT peptide.Application of AID-TAT peptide significantly decreased infarct size and supported contractility up to 12 weeks postcoronary artery occlusion as a result of a decrease in metabolic demand during reperfusion.

View Article: PubMed Central - PubMed

Affiliation: School of Anatomy, Physiology and Human Biology, The University of Western Australia, Crawley, WA, Australia (H.M.V., L.C.H.).

Show MeSH
Related in: MedlinePlus