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Upregulation of glycolytic enzymes, mitochondrial dysfunction and increased cytotoxicity in glial cells treated with Alzheimer's disease plasma.

Jayasena T, Poljak A, Braidy N, Smythe G, Raftery M, Hill M, Brodaty H, Trollor J, Kochan N, Sachdev P - PLoS ONE (2015)

Bottom Line: AD plasma and was found to significantly decrease cell viability and increase glycolytic flux in microglia compared to plasma from healthy controls.This effect was prevented by the heat inactivation of complement.Proteomic methods and isobaric tags (iTRAQ) found the expression level of complement and other acute phase proteins to be altered in MCI and AD plasma and an upregulation of key enzymes involved in the glycolysis pathway in cells exposed to AD plasma.

View Article: PubMed Central - PubMed

Affiliation: Bioanalytical Mass Spectrometry Facility, MW Analytical Centre, University of New South Wales, Sydney, Australia; Centre for Healthy Brain Ageing, School of Psychiatry, University of New South Wales, Sydney, Australia.

ABSTRACT
Alzheimer's disease (AD) is a neurodegenerative disorder associated with increased oxidative stress and neuroinflammation. Markers of increased protein, lipid and nucleic acid oxidation and reduced activities of antioxidant enzymes have been reported in AD plasma. Amyloid plaques in the AD brain elicit a range of reactive inflammatory responses including complement activation and acute phase reactions, which may also be reflected in plasma. Previous studies have shown that human AD plasma may be cytotoxic to cultured cells. We investigated the effect of pooled plasma (n = 20 each) from healthy controls, individuals with amnestic mild cognitive impairment (aMCI) and Alzheimer's disease (AD) on cultured microglial cells. AD plasma and was found to significantly decrease cell viability and increase glycolytic flux in microglia compared to plasma from healthy controls. This effect was prevented by the heat inactivation of complement. Proteomic methods and isobaric tags (iTRAQ) found the expression level of complement and other acute phase proteins to be altered in MCI and AD plasma and an upregulation of key enzymes involved in the glycolysis pathway in cells exposed to AD plasma. Altered expression levels of acute phase reactants in AD plasma may alter the energy metabolism of glia.

No MeSH data available.


Related in: MedlinePlus

Chromatogram of fractionation using Hu6 column and 1D SDS/PAGE of these fractions.Low abundant proteins are eluted first (first peak on chromatogram) and high abundant proteins are eluted after (second peak). Gel shows significant depletion of high abundant proteins in the low abundant fractions. Loading was 50 μg/lane. First and last lanes contained molecular weight markers. Each fraction was run in duplicate.
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pone.0116092.g003: Chromatogram of fractionation using Hu6 column and 1D SDS/PAGE of these fractions.Low abundant proteins are eluted first (first peak on chromatogram) and high abundant proteins are eluted after (second peak). Gel shows significant depletion of high abundant proteins in the low abundant fractions. Loading was 50 μg/lane. First and last lanes contained molecular weight markers. Each fraction was run in duplicate.

Mentions: Fractionation using the MARS-Hu6 column provided a baseline separation of low and high abundant proteins (Fig. 3). These fractions were run on a 1D SDS NuPage gel and proteins were shown to be effectively separated with a substantial depletion of high abundant proteins, revealing many lower abundant protein bands in the low abundant fraction (Fig. 3).


Upregulation of glycolytic enzymes, mitochondrial dysfunction and increased cytotoxicity in glial cells treated with Alzheimer's disease plasma.

Jayasena T, Poljak A, Braidy N, Smythe G, Raftery M, Hill M, Brodaty H, Trollor J, Kochan N, Sachdev P - PLoS ONE (2015)

Chromatogram of fractionation using Hu6 column and 1D SDS/PAGE of these fractions.Low abundant proteins are eluted first (first peak on chromatogram) and high abundant proteins are eluted after (second peak). Gel shows significant depletion of high abundant proteins in the low abundant fractions. Loading was 50 μg/lane. First and last lanes contained molecular weight markers. Each fraction was run in duplicate.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4364672&req=5

pone.0116092.g003: Chromatogram of fractionation using Hu6 column and 1D SDS/PAGE of these fractions.Low abundant proteins are eluted first (first peak on chromatogram) and high abundant proteins are eluted after (second peak). Gel shows significant depletion of high abundant proteins in the low abundant fractions. Loading was 50 μg/lane. First and last lanes contained molecular weight markers. Each fraction was run in duplicate.
Mentions: Fractionation using the MARS-Hu6 column provided a baseline separation of low and high abundant proteins (Fig. 3). These fractions were run on a 1D SDS NuPage gel and proteins were shown to be effectively separated with a substantial depletion of high abundant proteins, revealing many lower abundant protein bands in the low abundant fraction (Fig. 3).

Bottom Line: AD plasma and was found to significantly decrease cell viability and increase glycolytic flux in microglia compared to plasma from healthy controls.This effect was prevented by the heat inactivation of complement.Proteomic methods and isobaric tags (iTRAQ) found the expression level of complement and other acute phase proteins to be altered in MCI and AD plasma and an upregulation of key enzymes involved in the glycolysis pathway in cells exposed to AD plasma.

View Article: PubMed Central - PubMed

Affiliation: Bioanalytical Mass Spectrometry Facility, MW Analytical Centre, University of New South Wales, Sydney, Australia; Centre for Healthy Brain Ageing, School of Psychiatry, University of New South Wales, Sydney, Australia.

ABSTRACT
Alzheimer's disease (AD) is a neurodegenerative disorder associated with increased oxidative stress and neuroinflammation. Markers of increased protein, lipid and nucleic acid oxidation and reduced activities of antioxidant enzymes have been reported in AD plasma. Amyloid plaques in the AD brain elicit a range of reactive inflammatory responses including complement activation and acute phase reactions, which may also be reflected in plasma. Previous studies have shown that human AD plasma may be cytotoxic to cultured cells. We investigated the effect of pooled plasma (n = 20 each) from healthy controls, individuals with amnestic mild cognitive impairment (aMCI) and Alzheimer's disease (AD) on cultured microglial cells. AD plasma and was found to significantly decrease cell viability and increase glycolytic flux in microglia compared to plasma from healthy controls. This effect was prevented by the heat inactivation of complement. Proteomic methods and isobaric tags (iTRAQ) found the expression level of complement and other acute phase proteins to be altered in MCI and AD plasma and an upregulation of key enzymes involved in the glycolysis pathway in cells exposed to AD plasma. Altered expression levels of acute phase reactants in AD plasma may alter the energy metabolism of glia.

No MeSH data available.


Related in: MedlinePlus