Michalak 2017 Abstract MITOEAGLE Barcelona
Mitochonrial respiration in intact peripheral blood mononuclear cells in patients with extrapyramidal syndromes.
Event: MitoEAGLE Barcelona 2017
Mitochondrial dysfunction was suggested as a unifying pathophysiological explanation for Parkinson's disease and other neurodegenerative disorders. Mitochondria are the most efficient producers of ATP and play an important role in calcium homeostasis as well as in apoptosis initiation. In the case of severe oxidative phosphorylation defects, the ability of mitochondria to handle calcium is impaired . Moreover, mitochondria are a major source of reactive oxygen species (ROS) and contain defence mechanisms against ROS-mediated damage, including superoxide dismutase (SOD). Hence, mitochondria and oxidative stress are implicated in a number of neurodegenerative and aging – related disorders. Genes associated with autosomal recessive Parkinson’s disease are involved in the control of mitochondrial function. Phosphatase and Tensin homolog deleted on chromosome ten (PTEN) Induced Putative Kinase 1 (PINK1) and Parkin genes control the elimination of impaired mitochondria via autophagy (mitophagy). Alpha – synuclein, a presynaptic protein aggregates and produces intraneuronal protein inclusions called Lewy bodies. The role of alpha – synuclein in the pathophysiology of Parkinson’s disease awaits elucidation and it seems to be dichotomous. Alpha-synuclein can participate in defense mechanisms preserving neural mitochondrial homeostasis against oxidative stress, while not protecting against stressors directly affecting mitochondrial function .
With this background in mind we have undertaken the study on mitochonrial respiration in intact peripheral blood mononuclear cells in patients with extrapyramidal syndromes.
The study included 57 patients with extrapyramidal disorders. Idiopathic Parkinson’s disease (PD) was represented by 38 patients, progresive supranuclear palsy (PSP, Steele-Richardson-Olszewski’s disease) - 6 patients, cortico-basal degeneration (CDB) - 3 patients, and multi-system atrophy (MSA) - 9 patients and diffuse Lewy body disease (DLBD) - 1 patient. In contrast to idiopathic Parkinson’s disease, other extrapyramidal disorders like PSP, CBD, MSA and DLBD, were considered as atypical parkinsonisms. Extrapyramidal syndromes associated with alpha-synuclein pathology (PD, MSA, DLBD) were classified as synucleinopathies, other, like PSP and CBD were classified as tauopathies. Ten healthy volunteers were used as controls. Peripheral blood mononuclear cells (PBMC) were isolated from ethylenediamine tetraacetic acid (EDTA) blood by density gradient centrifugation (Histopaque, Sigma-Aldrich). The isolated fractions were supplemented with protease inhibitors cocktail (Sigma-Aldrich) on ice. The cell number was counted in Burker’s chamber and the volume corresponding to 1 000 000 cells was applied for respirometry. Cell viability was tested by means of trypan blue method, mitochondrial respiration was analysed in intact PBMCs according to ROUTINE, LEAK, ET-pathway, ROX protocol . The data abstracted from medical history were: the duration of disease, the presence of motor fluctuations and dyskinesias, comorbidities and treatment. All patients underwent standard neurological examination. Ultrasound examination of substatia nigra echogenicity and the diameter of 3rd ventricle was performed by using ALOKA Prosound Alpha 7 device according to standard procedure.
Labels: MiParea: Respiration Pathology: Aging;senescence, Neurodegenerative, Other
Organism: Human Tissue;cell: Blood cells Preparation: Intact cells
Coupling state: LEAK, ROUTINE, OXPHOS, ET Pathway: ROX HRR: Oxygraph-2k Event: A4 PMBCs
- Michalak S (1), Florczak J(2), Rybacka-Mossakowska J(1), Ambrosius W(2), Wysocka E(3), Osztynowicz K(1), Kozubski W(2)
- Dept Neurochem Neuropathol,
- Dept Neurology,
- Dept Clinical Biochemistry and Laboratory Medicine; Poznan Univ Medical Sciences, Poland. - [email protected]
We have found that ROUTINE respiration in PBMCs is decreased (P = 0.0237) in patients with extrapyramidal disorders (15.3±4.5 pmol/s*106 cells [mean±SD]) compared to healthy controls (19.2±4.4 pmol/s*106 cells). LEAK (P = 0.0330) and ROX (P = 0.0486) were also lower in subjects with extrapyramidal diseases (4.2; 2.98–5.68 [median; interquartile range] and 4.2± 2.0 pmol/s*106 cells, respectively) than in controls (6.0; 5.37 – 6.78 and 5.7± 1.6 pmol/s*106 cells, respectively). No differences in mitochondrial respiration in PBMCs were observed neither between PD and atypical parkinsonism patients, nor between patients with synucleinopathies or tauopathies. ROUTINE respiration was inhibited (P = 0.007) in PD patients with hyperechogenicity of substantia nigra in ultrasound (14.10 ± 3.6 pmol/s*106 cells) compared to controls (19.2 ± 4.4 pmol/s*106 cells). LEAK was decreased (P = 0,0203) in patients with idiopathic Parkinson's disease and hyperechogenicity of substantia nigra in ultasound (3.4; 2.1 – 5.2 pmol/s*106 cells) than in healthy controls (6.0; 5.37 – 6.78 pmol/s*106 cells). ROUTINE respiration and ROX correlated negatively with the diameter of 3rd ventricle measured in ultrasound (r= -0.3820; P = 0.0373 and r= -0.3969; P = 0.0330, respectively). ROX correlated negatively with the age of patients with extrapyramidal disorders (r= -0.2724; P = 0.0463). We have not found any effects of treatment with levodopa, dopaminę receptors agonists, selegiline, rasagiline and amantadine on mitochondrial respiration in intact PBMCs from the patients with extrapyramidal disorders.
To conclude, idiopathic Parkinson’s disease is associated with impaired mitochondrial respiration in intact PBMCs. The activity of respiratory states in intact PBMCs correlates with neurodegeneration measures in ultrasound examination of Parkinson’s disease patients. The treatment of extrapyramidal syndromes has no effect on mitochonrial respiration in PBMCs.
- von Kleist-Retzow JC, Hornig-Do HT, Schauen M, Eckertz S, Dinh TA, Stassen F, Lottmann N, Bust M, Galunska B, Wielckens K, Hein W, Beuth J, Braun JM, Fischer JH, Ganitkevich VY, Maniura- Weber K, Wiesner RJ (2007) Impaired mitochondrial Ca2+ homeostasis in respiratory chain-deficienT cells but efficient compensation of energetic disadvantage by enhanced anaerobic glycolysis due to low ATP steady state levels. Exp Cell Res 313:3076–89.
- Pickrell AM, Youle RJ (2015) The roles of PINK1, parkin, and mitochondrial fidelity in Parkinson’s disease. Neuron 85:257–73.
- Menges S, Minakaki G, Schaefer PM, Meixner H, Prots I, Schlötzer-Schrehardt U, Friedland K, Winner B, Outeiro TF, Winklhofer KF, von Arnim CA, Xiang W, Winkler J, Klucken J (2017) Alpha-synuclein prevents the formation of spherical mitochondria and apoptosis under oxidative stress. Sci Rep 7:42942.
- Gnaiger E, Renner-Sattler K (2014) High-resolution respirometry and coupling control protocol with intact cells: ROUTINE, LEAK, ET-pathway, ROX. Mitochondrial Physiology Network 08.09:1-8.