Methylene blue prevents neurodegeneration caused by rotenone in the retina

An experimental optic neuropathy model was used to test the hypothesis that methylene blue may protect the retinal ganglion cell layer from neurodegeneration caused by rotenone. Rotenone is a widely used pesticide that inhibits complex I, the first enzyme of the mitochondrial respiratory chain. Complex I dysfunction is linked to the degeneration of retinal ganglion cells in Leber’s optic neuropathy. Methylene blue is a reduction-oxidation agent that can act as a powerful antioxidant and also as an enhancer of the electron transport chain, preventing formation of mitochondrial oxygen free radicals and promoting oxygen consumption. The neurodegeneration of the retina was studied in mice with intravitreal microinjection of rotenone alone, or in combination with increasing doses of methylene blue, in one eye, and the vehicle in the contralateral control eye. The effect of rotenone and rotenone plus methylene blue was investigated using two histological stains, complex I and Nissl, and two measurements, morphometric layer thickness and non-biased stereological cell counts. Rotenone induced neurodegeneration in the retinal ganglion cell layer 24 h after injection, as indicated by significant reductions in both the thickness and cell numbers of the retinal ganglion cell layer of eyes microinjected with rotenone as compared to the control eyes. This neurodegeneration was prevented in a dose dependent manner by the injection of methylene blue along with rotenone. It was concluded that rotenone-induced degeneration in the ganglion cell layer can be prevented by intravitreal injection of methylene blue.In vitro experiments showed that methylene blue is both a powerful antioxidant as well as an enhancer of cellular oxygen consumption and is able to reverse the oxidative stress and decrease in oxygen consumption induced by rotenone in brain homogenates. The findings suggest that methylene blue may be a promising neuroprotective agent in optic neuropathy and perhaps other neurodegenerative diseases caused by mitochondrial dysfunction.

[1]  Simon R. Cherry,et al.  Effects of Image Resolution on Autoradiographic Measurements of Posterior Cingulate Activity in PDAPP Mice: Implications for Functional Brain Imaging Studies of Transgenic Mouse Models of Alzheimer's Disease , 2002, NeuroImage.

[2]  F. Gonzalez-Lima,et al.  Cytochrome Oxidase Inhibition In Alzheimer’s Disease , 1998 .

[3]  K. C. Richardson,et al.  The fine structure of autonomic nerves after vital staining with methylene blue , 1969, The Anatomical record.

[4]  James L. McGaugh,et al.  Methylene blue alters retention of inhibitory avoidance responses , 1978 .

[5]  T. Cerny,et al.  Methylene blue and the neurotoxic mechanisms of ifosfamide encephalopathy , 1996, European Journal of Clinical Pharmacology.

[6]  F. Gonzalez-Lima,et al.  Cytochrome oxidase activity in the auditory system of the mouse: A qualitative and quantitative histochemical study , 1994, Neuroscience.

[7]  M. Esposti.  Inhibitors of NADH-ubiquinone reductase: an overview. , 1998 .

[8]  S. Bradberry.  Occupational Methaemoglobinaemia , 2003, Toxicological reviews.

[9]  F. Gonzalez-Lima,et al.  Mouse model of optic neuropathy caused by mitochondrial complex I dysfunction , 2002, Neuroscience Letters.

[10]  D. Choi,et al.  Glutamate neurotoxicity and diseases of the nervous system , 1988, Neuron.

[11]  U. Dräger,et al.  Ganglion cell distribution in the retina of the mouse. , 1981, Investigative ophthalmology & visual science.

[12]  Todd B. Sherer,et al.  Chronic systemic pesticide exposure reproduces features of Parkinson's disease , 2000, Nature Neuroscience.

[13]  I. Fridovich,et al.  Intracellular production of superoxide radical and of hydrogen peroxide by redox active compounds. , 1979, Archives of biochemistry and biophysics.

[14]  M. Wainwright,et al.  Methylene Blue - a Therapeutic Dye for All Seasons? , 2002, Journal of chemotherapy.

[15]  B. Mayer,et al.  Inhibition of nitric oxide synthesis by methylene blue. , 1993, Biochemical pharmacology.

[16]  G. Naylor,et al.  A two-year double-blind crossover trial of the prophylactic effect of methylene blue in manicdepressive psychosis , 1986, Biological Psychiatry.

[17]  S M Podos,et al.  Elevated glutamate levels in the vitreous body of humans and monkeys with glaucoma. , 1996, Archives of ophthalmology.

[18]  R. Ramsay,et al.  The reaction sites of rotenone and ubiquinone with mitochondrial NADH dehydrogenase. , 1994, Biochimica et biophysica acta.

[19]  M. Wong-Riley.  Cytochrome oxidase: an endogenous metabolic marker for neuronal activity , 1989, Trends in Neurosciences.

[20]  Dirk Jones,et al.  Quantitative mapping of cytovhrome oxidase activity in the central auditory system of the gerbil: a study with calibrated activity standards and metal-intensified histochemistry , 1994, Brain Research.

[21]  F Gonzalez-Lima,et al.  Memory facilitation by methylene blue: dose-dependent effect on behavior and brain oxygen consumption. , 2005, European journal of pharmacology.

[22]  N. Howell.  Leber hereditary optic neuropathy: respiratory chain dysfunction and degeneration of the optic nerve , 1998, Vision Research.

[23]  M. Beal,et al.  Mitochondrial dysfunction in neurodegenerative diseases. , 1998, Biochimica et biophysica acta.

[24]  J. Borowitz,et al.  NMDA Receptor Activation Produces Concurrent Generation of Nitric Oxide and Reactive Oxygen Species: Implications for Cell Death , 1995, Journal of neurochemistry.

[25]  J. Bockaert,et al.  NMDA-dependent superoxide production and neurotoxicity , 1993, Nature.

[26]  A. Schapira,et al.  Human complex I defects in neurodegenerative diseases. , 1998, Biochimica et biophysica acta.

[27]  D. Wallace,et al.  Leber's hereditary optic neuropathy: a model for mitochondrial neurodegenerative diseases , 1992, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[28]  F. Gonzalez-Lima,et al.  Quantitative Histochemistry of Cytochrome Oxidase Activity , 1998 .

[29]  P. Agostinho,et al.  Glutamate in life and death of retinal amacrine cells. , 1998, General pharmacology.

[30]  J. Coyle,et al.  Oxidative stress, glutamate, and neurodegenerative disorders. , 1993, Science.

[31]  Zuoshang Xu,et al.  A quantitative histochemical assay for activities of mitochondrial electron transport chain complexes in mouse spinal cord sections , 2002, Journal of Neuroscience Methods.

[32]  B. Lauterburg,et al.  Pharmacokinetics and organ distribution of intravenous and oral methylene blue , 2000, European Journal of Clinical Pharmacology.

[33]  K. Davies,et al.  The oxidative inactivation of mitochondrial electron transport chain components and ATPase. , 1990, The Journal of biological chemistry.

[34]  T. Tseng,et al.  Hibiscus protocatechuic acid or esculetin can inhibit oxidative LDL induced by either copper ion or nitric oxide donor. , 2002, Journal of agricultural and food chemistry.

[35]  J Inukai,et al.  Supravital staining of mammalian brain with intra-arterial methylene blue followed by pressurized oxygen. , 1968, Stain technology.

[36]  David L. McCollum,et al.  Methylene blue restores spatial memory retention impaired by an inhibitor of cytochrome oxidase in rats , 2002, Neuroscience Letters.

[37]  F. Gonzalez-Lima,et al.  Methylene blue improves brain oxidative metabolism and memory retention in rats , 2004, Pharmacology Biochemistry and Behavior.

[38]  J. Stucki,et al.  Stimulation of respiration by methylene blue in rat liver mitochondria , 1997, FEBS letters.

[39]  Mark P. Mattson,et al.  Excitotoxic and excitoprotective mechanisms , 2007, NeuroMolecular Medicine.

[40]  J. Cooper,et al.  Irreversible Inhibition of Mitochondrial Complex I by 1‐Methyl‐4‐Phenylpyridinium: Evidence for Free Radical Involvement , 1992, Journal of neurochemistry.

[41]  M. Kelner,et al.  Potential of methylene blue to block oxygen radical generation in reperfusion injury. , 1988, Basic life sciences.

[42]  F. Gonzalez-Lima,et al.  Quantitative cytochemistry of cytochrome oxidase and cellular morphometry of the human inferior colliculus in control and Alzheimer's patients , 1997, Brain Research.

[43]  G. Cole,et al.  Inhibition of caspase-3-like activity reduces glutamate induced cell death in adult rat retina 1 1 Published on the World Wide Web on 9 May 2001. , 2001, Brain Research.

[44]  C. Babbs,et al.  Methylene blue as an inhibitor of superoxide generation by xanthine oxidase. A potential new drug for the attenuation of ischemia/reperfusion injury. , 1991, Biochemical pharmacology.

[45]  T H Shepard,et al.  National toxicology program. , 1981, Teratology.

[46]  J. Nobrega,et al.  Long-term changes in regional brain cytochrome oxidase activity induced by electroconvulsive treatment in rats , 1993, Brain Research.

[47]  Todd B. Sherer,et al.  Subcutaneous Rotenone Exposure Causes Highly Selective Dopaminergic Degeneration and α-Synuclein Aggregation , 2003, Experimental Neurology.

[48]  D. Choi.  Calcium: still center-stage in hypoxic-ischemic neuronal death , 1995, Trends in Neurosciences.

[49]  P E LINDAHL,et al.  The effect of rotenone on respiration and its point of attack. , 1961, Experimental cell research.