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Analysis of Mitochondrial haemoglobin in Parkinson's disease brain.

Shephard F, Greville-Heygate O, Liddell S, Emes R, Chakrabarti L - Mitochondrion (2016)

Bottom Line: We hypothesised that altered physiological processes are associated with recruitment and localisation of haemoglobin to these organelles.These new data illustrate dynamic localisation of mitochondrial haemoglobin within the cell.It has been postulated that cerebellar circuitry may be activated to play a protective role in individuals with Parkinson's.

View Article: PubMed Central - PubMed

Affiliation: University of Nottingham, Faculty of Medicine, SVMS, Sutton Bonington Campus, LE12 5RD, England, UK.

No MeSH data available.


Related in: MedlinePlus

A. Mitochondrial HbA migrates from the intermembrane space to the outer membrane in affected human male cerebellum. Mitochondrial samples were sub-fractioned to allow examination of HbA localisation within the organelle, a representative gel is shown for each gender and control (total n = 8). The male Parkinson's disease brain demonstrated a shift in HbA localisation from the intermembrane space to the outer membrane fraction. This was not seen in the control or female Parkinson's brain mitochondria. Levels of HbA in the IMS were quantified in control and PD samples for both male and female patients (n = 2 for each), using Image J. Please see Supplemental Fig. 7 for all gel images.Levels of HbA in the IMS were significantly decreased in male PD compared with male control (p = 0.028 using unpaired two-tailed t-test). No significant change in female PD compared with female control (p $_amp_$gt; 0.05, unpaired two-tailed t-test).Mito fr – mitochondrial fraction, OM – outer membrane, IMS – inter membrane space, IM – inner membrane, M - matrix.B. Cycles of hypoxia result in increased hb in Drosophila mitochondrial fractions. Mitochondrial/cytoplasmic Hb levels determined using Western blotting, normalised to beta-actin. Hypoxia conditions: 2.5% O2 30 min 25 °C followed by normoxia 30 min 25 °C (middle bar) 2.5% O2 30 min 25 °C followed by normoxia 30 min 25 °C × 2 (right hand bar). 40–100 flies per condition. n = 3, * p $_amp_$lt; 0.05 (1 tailed t-test).
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f0005: A. Mitochondrial HbA migrates from the intermembrane space to the outer membrane in affected human male cerebellum. Mitochondrial samples were sub-fractioned to allow examination of HbA localisation within the organelle, a representative gel is shown for each gender and control (total n = 8). The male Parkinson's disease brain demonstrated a shift in HbA localisation from the intermembrane space to the outer membrane fraction. This was not seen in the control or female Parkinson's brain mitochondria. Levels of HbA in the IMS were quantified in control and PD samples for both male and female patients (n = 2 for each), using Image J. Please see Supplemental Fig. 7 for all gel images.Levels of HbA in the IMS were significantly decreased in male PD compared with male control (p = 0.028 using unpaired two-tailed t-test). No significant change in female PD compared with female control (p $_amp_$gt; 0.05, unpaired two-tailed t-test).Mito fr – mitochondrial fraction, OM – outer membrane, IMS – inter membrane space, IM – inner membrane, M - matrix.B. Cycles of hypoxia result in increased hb in Drosophila mitochondrial fractions. Mitochondrial/cytoplasmic Hb levels determined using Western blotting, normalised to beta-actin. Hypoxia conditions: 2.5% O2 30 min 25 °C followed by normoxia 30 min 25 °C (middle bar) 2.5% O2 30 min 25 °C followed by normoxia 30 min 25 °C × 2 (right hand bar). 40–100 flies per condition. n = 3, * p $_amp_$lt; 0.05 (1 tailed t-test).

Mentions: Sub-mitochondrial fractions were prepared from cerebella and interrogated for HbA content (Fig. 1A). Fractions were verified using appropriate antibodies SMAC and NDUFS3 for intermembrane space and inner membrane respectively. Control (male 80 years) and affected (male, 82 years, 18 years of disease) cerebella mitochondria were prepared and subfractionated. A female (85 years old, 18 years of disease) sub-fractionated sample is included for comparison. HbA content of the fractions was determined by western blotting. We found HbA to be present in the inter membrane space of the control sample which corroborates earlier findings (Shephard et al., 2014). In the affected sample there is little evidence of HbA in the inter membrane space. The HbA in the affected sample appears in the outer membrane fraction. Though the amount of HbA was not absolutely quantified, relative ratios suggest that there is a substantial quantity of this protein in or associated with the outer membrane of the affected sample mitochondria.


Analysis of Mitochondrial haemoglobin in Parkinson's disease brain.

Shephard F, Greville-Heygate O, Liddell S, Emes R, Chakrabarti L - Mitochondrion (2016)

A. Mitochondrial HbA migrates from the intermembrane space to the outer membrane in affected human male cerebellum. Mitochondrial samples were sub-fractioned to allow examination of HbA localisation within the organelle, a representative gel is shown for each gender and control (total n = 8). The male Parkinson's disease brain demonstrated a shift in HbA localisation from the intermembrane space to the outer membrane fraction. This was not seen in the control or female Parkinson's brain mitochondria. Levels of HbA in the IMS were quantified in control and PD samples for both male and female patients (n = 2 for each), using Image J. Please see Supplemental Fig. 7 for all gel images.Levels of HbA in the IMS were significantly decreased in male PD compared with male control (p = 0.028 using unpaired two-tailed t-test). No significant change in female PD compared with female control (p $_amp_$gt; 0.05, unpaired two-tailed t-test).Mito fr – mitochondrial fraction, OM – outer membrane, IMS – inter membrane space, IM – inner membrane, M - matrix.B. Cycles of hypoxia result in increased hb in Drosophila mitochondrial fractions. Mitochondrial/cytoplasmic Hb levels determined using Western blotting, normalised to beta-actin. Hypoxia conditions: 2.5% O2 30 min 25 °C followed by normoxia 30 min 25 °C (middle bar) 2.5% O2 30 min 25 °C followed by normoxia 30 min 25 °C × 2 (right hand bar). 40–100 flies per condition. n = 3, * p $_amp_$lt; 0.05 (1 tailed t-test).
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f0005: A. Mitochondrial HbA migrates from the intermembrane space to the outer membrane in affected human male cerebellum. Mitochondrial samples were sub-fractioned to allow examination of HbA localisation within the organelle, a representative gel is shown for each gender and control (total n = 8). The male Parkinson's disease brain demonstrated a shift in HbA localisation from the intermembrane space to the outer membrane fraction. This was not seen in the control or female Parkinson's brain mitochondria. Levels of HbA in the IMS were quantified in control and PD samples for both male and female patients (n = 2 for each), using Image J. Please see Supplemental Fig. 7 for all gel images.Levels of HbA in the IMS were significantly decreased in male PD compared with male control (p = 0.028 using unpaired two-tailed t-test). No significant change in female PD compared with female control (p $_amp_$gt; 0.05, unpaired two-tailed t-test).Mito fr – mitochondrial fraction, OM – outer membrane, IMS – inter membrane space, IM – inner membrane, M - matrix.B. Cycles of hypoxia result in increased hb in Drosophila mitochondrial fractions. Mitochondrial/cytoplasmic Hb levels determined using Western blotting, normalised to beta-actin. Hypoxia conditions: 2.5% O2 30 min 25 °C followed by normoxia 30 min 25 °C (middle bar) 2.5% O2 30 min 25 °C followed by normoxia 30 min 25 °C × 2 (right hand bar). 40–100 flies per condition. n = 3, * p $_amp_$lt; 0.05 (1 tailed t-test).
Mentions: Sub-mitochondrial fractions were prepared from cerebella and interrogated for HbA content (Fig. 1A). Fractions were verified using appropriate antibodies SMAC and NDUFS3 for intermembrane space and inner membrane respectively. Control (male 80 years) and affected (male, 82 years, 18 years of disease) cerebella mitochondria were prepared and subfractionated. A female (85 years old, 18 years of disease) sub-fractionated sample is included for comparison. HbA content of the fractions was determined by western blotting. We found HbA to be present in the inter membrane space of the control sample which corroborates earlier findings (Shephard et al., 2014). In the affected sample there is little evidence of HbA in the inter membrane space. The HbA in the affected sample appears in the outer membrane fraction. Though the amount of HbA was not absolutely quantified, relative ratios suggest that there is a substantial quantity of this protein in or associated with the outer membrane of the affected sample mitochondria.

Bottom Line: We hypothesised that altered physiological processes are associated with recruitment and localisation of haemoglobin to these organelles.These new data illustrate dynamic localisation of mitochondrial haemoglobin within the cell.It has been postulated that cerebellar circuitry may be activated to play a protective role in individuals with Parkinson's.

View Article: PubMed Central - PubMed

Affiliation: University of Nottingham, Faculty of Medicine, SVMS, Sutton Bonington Campus, LE12 5RD, England, UK.

No MeSH data available.


Related in: MedlinePlus