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Alcohol-related brain damage in humans.

Erdozain AM, Morentin B, Bedford L, King E, Tooth D, Brewer C, Wayne D, Johnson L, Gerdes HK, Wigmore P, Callado LF, Carter WG - PLoS ONE (2014)

Bottom Line: BA 9 tissue homogenisation and one dimensional polyacrylamide gel electrophoresis (PAGE) proteomics of cytosolic proteins identified dramatic reductions in the protein levels of spectrin β II, and α- and β-tubulins in alcoholics, and these were validated and quantitated by Western blotting.There was also a significant reduction in proteasome activity in alcoholics.One dimensional PAGE of membrane-enriched fractions detected a reduction in β-spectrin protein levels, and a significant increase in transmembranous α3 (catalytic) subunit of the Na+,K+-ATPase in alcoholic subjects.

View Article: PubMed Central - PubMed

Affiliation: School of Medicine, University of Nottingham, Royal Derby Hospital Centre, Derby, United Kingdom; Department of Pharmacology, University of the Basque Country, and Centro de Investigación Biomédica en Red de Salud Mental, Spain.

ABSTRACT
Chronic excessive alcohol intoxications evoke cumulative damage to tissues and organs. We examined prefrontal cortex (Brodmann's area (BA) 9) from 20 human alcoholics and 20 age, gender, and postmortem delay matched control subjects. H & E staining and light microscopy of prefrontal cortex tissue revealed a reduction in the levels of cytoskeleton surrounding the nuclei of cortical and subcortical neurons, and a disruption of subcortical neuron patterning in alcoholic subjects. BA 9 tissue homogenisation and one dimensional polyacrylamide gel electrophoresis (PAGE) proteomics of cytosolic proteins identified dramatic reductions in the protein levels of spectrin β II, and α- and β-tubulins in alcoholics, and these were validated and quantitated by Western blotting. We detected a significant increase in α-tubulin acetylation in alcoholics, a non-significant increase in isoaspartate protein damage, but a significant increase in protein isoaspartyl methyltransferase protein levels, the enzyme that triggers isoaspartate damage repair in vivo. There was also a significant reduction in proteasome activity in alcoholics. One dimensional PAGE of membrane-enriched fractions detected a reduction in β-spectrin protein levels, and a significant increase in transmembranous α3 (catalytic) subunit of the Na+,K+-ATPase in alcoholic subjects. However, control subjects retained stable oligomeric forms of α-subunit that were diminished in alcoholics. In alcoholics, significant loss of cytosolic α- and β-tubulins were also seen in caudate nucleus, hippocampus and cerebellum, but to different levels, indicative of brain regional susceptibility to alcohol-related damage. Collectively, these protein changes provide a molecular basis for some of the neuronal and behavioural abnormalities attributed to alcoholics.

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Western blot of α- and β-tubulins in nuclear and membrane fractions from the prefrontal cortex of control and alcoholic subjects.Control (C) or alcoholic (A) prefrontal cortex nuclear and membrane fractions were resolved by 1D PAGE and proteins Western blotted for α- and β-tubulins.
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pone-0093586-g004: Western blot of α- and β-tubulins in nuclear and membrane fractions from the prefrontal cortex of control and alcoholic subjects.Control (C) or alcoholic (A) prefrontal cortex nuclear and membrane fractions were resolved by 1D PAGE and proteins Western blotted for α- and β-tubulins.

Mentions: We also examined the levels of α- and β-tubulin within nuclear and membrane fractions to assess whether the reduced cytosolic protein levels seen in alcoholics arose from protein translocation to these other cellular compartments. After 1D PAGE and Western blotting there were no significant changes in the levels of α- or β-tubulins in the nuclear or membrane fractions of alcoholic brain tissue when compared to control subjects – Figure 4. This suggested that α and β-tubulin protein loss in the prefrontal cortex of alcoholics was limited to the cytosol.


Alcohol-related brain damage in humans.

Erdozain AM, Morentin B, Bedford L, King E, Tooth D, Brewer C, Wayne D, Johnson L, Gerdes HK, Wigmore P, Callado LF, Carter WG - PLoS ONE (2014)

Western blot of α- and β-tubulins in nuclear and membrane fractions from the prefrontal cortex of control and alcoholic subjects.Control (C) or alcoholic (A) prefrontal cortex nuclear and membrane fractions were resolved by 1D PAGE and proteins Western blotted for α- and β-tubulins.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3974765&req=5

pone-0093586-g004: Western blot of α- and β-tubulins in nuclear and membrane fractions from the prefrontal cortex of control and alcoholic subjects.Control (C) or alcoholic (A) prefrontal cortex nuclear and membrane fractions were resolved by 1D PAGE and proteins Western blotted for α- and β-tubulins.
Mentions: We also examined the levels of α- and β-tubulin within nuclear and membrane fractions to assess whether the reduced cytosolic protein levels seen in alcoholics arose from protein translocation to these other cellular compartments. After 1D PAGE and Western blotting there were no significant changes in the levels of α- or β-tubulins in the nuclear or membrane fractions of alcoholic brain tissue when compared to control subjects – Figure 4. This suggested that α and β-tubulin protein loss in the prefrontal cortex of alcoholics was limited to the cytosol.

Bottom Line: BA 9 tissue homogenisation and one dimensional polyacrylamide gel electrophoresis (PAGE) proteomics of cytosolic proteins identified dramatic reductions in the protein levels of spectrin β II, and α- and β-tubulins in alcoholics, and these were validated and quantitated by Western blotting.There was also a significant reduction in proteasome activity in alcoholics.One dimensional PAGE of membrane-enriched fractions detected a reduction in β-spectrin protein levels, and a significant increase in transmembranous α3 (catalytic) subunit of the Na+,K+-ATPase in alcoholic subjects.

View Article: PubMed Central - PubMed

Affiliation: School of Medicine, University of Nottingham, Royal Derby Hospital Centre, Derby, United Kingdom; Department of Pharmacology, University of the Basque Country, and Centro de Investigación Biomédica en Red de Salud Mental, Spain.

ABSTRACT
Chronic excessive alcohol intoxications evoke cumulative damage to tissues and organs. We examined prefrontal cortex (Brodmann's area (BA) 9) from 20 human alcoholics and 20 age, gender, and postmortem delay matched control subjects. H & E staining and light microscopy of prefrontal cortex tissue revealed a reduction in the levels of cytoskeleton surrounding the nuclei of cortical and subcortical neurons, and a disruption of subcortical neuron patterning in alcoholic subjects. BA 9 tissue homogenisation and one dimensional polyacrylamide gel electrophoresis (PAGE) proteomics of cytosolic proteins identified dramatic reductions in the protein levels of spectrin β II, and α- and β-tubulins in alcoholics, and these were validated and quantitated by Western blotting. We detected a significant increase in α-tubulin acetylation in alcoholics, a non-significant increase in isoaspartate protein damage, but a significant increase in protein isoaspartyl methyltransferase protein levels, the enzyme that triggers isoaspartate damage repair in vivo. There was also a significant reduction in proteasome activity in alcoholics. One dimensional PAGE of membrane-enriched fractions detected a reduction in β-spectrin protein levels, and a significant increase in transmembranous α3 (catalytic) subunit of the Na+,K+-ATPase in alcoholic subjects. However, control subjects retained stable oligomeric forms of α-subunit that were diminished in alcoholics. In alcoholics, significant loss of cytosolic α- and β-tubulins were also seen in caudate nucleus, hippocampus and cerebellum, but to different levels, indicative of brain regional susceptibility to alcohol-related damage. Collectively, these protein changes provide a molecular basis for some of the neuronal and behavioural abnormalities attributed to alcoholics.

Show MeSH
Related in: MedlinePlus