The use of phosphorescence oxygen analyzer to measure the effects of rotenone and 1-methyl-4-phenylpyridinium on striatal cellular respiration in C57BL6 mice

Mariam Al Shamsi; M Emdadul Haque; Allen Shahin; Sami Shaban; Abdul-Kader Souid

doi:10.1016/j.heliyon.2021.e07219

The use of phosphorescence oxygen analyzer to measure the effects of rotenone and 1-methyl-4-phenylpyridinium on striatal cellular respiration in C57BL6 mice

Heliyon. 2021 Jun 5;7(6):e07219. doi: 10.1016/j.heliyon.2021.e07219. eCollection 2021 Jun.

Authors

Mariam Al Shamsi¹, M Emdadul Haque², Allen Shahin¹, Sami Shaban³, Abdul-Kader Souid⁴

Affiliations

¹ Department of Microbiology and Immunology, UAE University, College of Medicine and Health Sciences, Alain, P.O. Box 17666, Abu Dhabi, United Arab Emirates.
² Department of Biochemistry and Molecular Biology, UAE University, College of Medicine and Health Sciences, Alain, P.O. Box 17666, Abu Dhabi, United Arab Emirates.
³ Department of Medical Education, UAE University, College of Medicine and Health Sciences, Alain, P.O. Box 17666, Abu Dhabi, United Arab Emirates.
⁴ Department of Pediatrics, UAE University, College of Medicine and Health Sciences, Alain, P.O. Box 17666, Abu Dhabi, United Arab Emirates.

Abstract

Background: We have previously reported on the use of the phosphorescence oxygen analyzer for measuring spinal cord cellular respiration. This analytical tool is used here to investigate the effects of two inhibitors of NADH:ubiquinone oxidoreductase, rotenone and 1-methyl-4-phenylpyridinium, on cellular respiration in striatal tissue. Both neurotoxins can induce Parkinson's disease-like symptoms, and have been used to study this disease in animals. Our hypothesis is that striatal cellular respiration is a sensitive biomarker for the adverse effects of toxins, and the phosphorescence oxygen analyzer can be used as a screening tool for this purpose.

Methods: Striatal fragments were collected from C57BL6 mice and immersed in Pd phosphor solution [phosphate-buffered saline, 3.0 μM 'Pd(II)-meso-tetra (sulfophenyl) tetrabenzoporphyrin' and 0.5% fat-free albumin, with and without 5.0 mM glucose]. The sample was transferred to a glass vial containing 2-mL Pd phosphor solution. The vial was sealed from air and placed in the instrument that measures dissolved oxygen as function of time. Immunoblots of the studied tissue were positive for the dopamine neuronal cell biomarker tyrosine hydroxylase.

Results: Striatal oxygen consumption was linear with time, exhibiting zero-order kinetics of oxygen reduction by cytochrome oxidase. Cyanide sensitive respiration was ≥90%, confirming oxygen was reduced by cytochrome oxidase. The rate of respiration increased by ~2-fold in the presence of glucose. Striatal oxygen consumption in the presence of rotenone or 1-methyl-4-phenylpyridinium was exponential, demonstrating impaired respiration.

Conclusion: Striatal cellular mitochondrial oxygen consumption was impaired by the studied inhibitors of complex I of the respiratory chain. This effect is expected to deplete NAD+ (oxidized nicotinamide adenine dinucleotide), a principle driver of glycolysis. In vivo studies are required to determine if these toxin-induced metabolic derangements contribute to the development of sporadic Parkinson's disease. This analytic tool can be used to screen environmental toxins for their in vitro effects on the striatum.

Keywords: Basal ganglia; Cellular bioenergetics; Cellular respiration; Neurons; Parkinson's disease; Substantia nigra.