THE DETERMINATION BY QUANTITATIVE HISTOCHEMISTRY OF THE EFFECT OF PHENOTHIAZINES ON BRAIN CYTOCHROME <i>C</i> OXIDASE ACTIVITY

Moraczewski, Albert S.; Anderson, Rae C.

doi:10.1177/14.1.64

Cited by 9 publications

(3 citation statements)

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“…Light and electron microscopic detection of cytochrome c oxidase (CCO) activity was conducted by the method of Moraczewski and Anderson11 with some modification. In brief, frozen sections were placed on glass or in 0.1 M phosphate buffer (pH 7.4) with 5% glucose, for light and electron microscopy, respectively.…”

Section: Methodsmentioning

confidence: 99%

Cadmium‐Induced Nephropathy in Rats Is Mediated by Expression of Senescence‐Associated Beta‐Galactosidase and Accumulation of Mitochondrial DNA Deletion

Takaki

Jimi

Segawa

et al. 2004

Annals of the New York Academy of Sciences

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Long-term exposure to cadmium (Cd) induces perturbation of kidney proximal tubular epithelial cells. Mitochondrial dysfunction in renal cortical cells may contribute to the pathogenesis of Cd-induced nephropathy. In this study, we examined the accumulation of mitochondrial DNA (mtDNA) with a large deletion and cellular senescence in the renal cortex. Wistar rats at 8 weeks of age were intraperitoneally injected with 1 mL of 1 mM CdCl(2) or saline, 3 times/week for 5, 20, 40, or 80 weeks. Mitochondrial Cd content in the renal cortex was quantified by atomic absorption analysis. Cytochrome c oxidase (CCO) and senescence-associated beta-galactosidase (SA-beta-gal) activity were determined in renal cortex by enzyme-histochemistry. mtDNA in total DNA extracted from the renal cortex was amplified by PCR, and mtDNA deletions, including 4,834-bp (nt8118-nt12937) deletion, were determined and semiquantified. After 40 weeks of Cd injection, Cd levels in the renal cortex reached a saturation level, and 30% of the level of the whole-cell fraction was found in the mitochondria. CCO activity in the renal cortex, which was predominantly found in proximal tubular cells, decreased after 40 weeks of Cd exposure. Expression of SA-beta-gal was detected primarily in the proximal tubular cells and significantly increased after 80 weeks of Cd exposure. After 40 weeks of study, accumulation of 4,834-bp deletion in mtDNA was evident in both groups of rats; however, the amount of the deletion was significantly greater in Cd-treated rats than in control rats. Our results indicate that long-term Cd exposure induced a post-regenerative state of proximal tubular cells, which accelerated accumulation of 4,834-bp mtDNA deletions in the renal cortex, suggesting that Cd may be a senescence acceleration factor for kidney proximal tubular epithelial cells, which results in Cd-induced nephropathy.

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Section: Methodsmentioning

confidence: 99%

Cadmium‐Induced Nephropathy in Rats Is Mediated by Expression of Senescence‐Associated Beta‐Galactosidase and Accumulation of Mitochondrial DNA Deletion

Takaki

Jimi

Segawa

et al. 2004

Annals of the New York Academy of Sciences

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“…Although brain CCO activity has been studied for over 40 years (Moraczewski & Anderson, 1966), advancements in techniques have been restrained by methods of sample preparation. Age-related differences in CCO activity observed in brain homogenates (Fattoretti et al , 2006) and synaptic mitochondria (Sen et al , 2007; Bertoni-Freddari et al , 2006) have, in the past, failed to differentiate between the aging neurons and the brain’s vasculature, hormones, and immune system.…”

Section: Introductionmentioning

confidence: 99%

Critical age-related loss of cofactors of neuron cytochrome C oxidase reversed by estrogen

Jones

Brewer

2009

Experimental Neurology

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The mechanistic basis for the correlation between mitochondrial dysfunction and neurodegenerative disease is unclear, but evidence supports involvement of cytochrome C oxidase (CCO) deficits with age. Neurons isolated from the brains of 24 month and 9 month rats and cultured in common conditions provide a model of intrinsic neuronal aging. In situ CCO activity was decreased in 24 month neurons relative to 9 month neurons. Possible CCO-related deficits include holoenzyme activity, cofactor, and substrate. No difference was found between neurons from 24 month and 9 month rats in mitochondrial counts per neuron, CCO activity in submitochondrial particles, or basal respiration. Immunostaining for cytochrome C in individual mitochondria revealed an age-related deficit of this electron donor. 24 month neurons did not have adequate respiratory capacity to upregulate respiration after a glutamate stimulus, in spite of a two-fold upregulation of respiration seen in 9 month neurons. Respiration in 24 month neurons was inhibited by lower concentrations of potassium cyanide, suggesting a 50% deficit in functional enzyme in 24 month compared to 9 month neurons. In addition to cytochrome C, CCO requires cardiolipin to function. Staining with nonylacridine orange revealed an age-related deficit in cardiolipin. Treatment of 24 month neurons with 17-β-estradiol restored cardiolipin levels (10 ng/mL) and upregulated respiration under glutamate stress (1 pg/mL). Attempts to induce mitochondrial turnover by neuronal multiplication also rejuvenated CCO activity in 24 month neurons. These data suggest cytochrome C and cardiolipin levels are deficient in 24 month neurons, preventing normal upregulation of respiration needed for oxidative phosphorylation in response to stress. Furthermore, the data suggest this deficit can be corrected with estrogen treatment.

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“…Light and electron microscopic detection of cytochrome c oxidase (COX) activity in Cd-treated and control rats (five rats at each week of study) was conducted by the method of Moraczewski and Anderson (1966) with some modifications. The isolated kidneys were fixed at 4 • C for 30 min in 0.1 M phosphate buffer (pH 7.4) containing 0.5% glutaraldehyde, 2% paraformaldehyde and 5% sucrose for electron microscopy or in 0.1 M phosphate buffer (pH 7.4) containing 2% paraformaldehyde and 5% sucrose for light microscopy.…”

Section: Enzyme-histochemistry Of Cytochrome C Oxidasementioning

confidence: 99%

Long-term cadmium exposure accelerates age-related mitochondrial changes in renal epithelial cells

et al. 2004

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THE DETERMINATION BY QUANTITATIVE HISTOCHEMISTRY OF THE EFFECT OF PHENOTHIAZINES ON BRAIN CYTOCHROME C OXIDASE ACTIVITY

Cited by 9 publications

References 0 publications

Cadmium‐Induced Nephropathy in Rats Is Mediated by Expression of Senescence‐Associated Beta‐Galactosidase and Accumulation of Mitochondrial DNA Deletion

Cadmium‐Induced Nephropathy in Rats Is Mediated by Expression of Senescence‐Associated Beta‐Galactosidase and Accumulation of Mitochondrial DNA Deletion

Critical age-related loss of cofactors of neuron cytochrome C oxidase reversed by estrogen

Long-term cadmium exposure accelerates age-related mitochondrial changes in renal epithelial cells

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