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Analysis of spatial and temporal protein expression in the cerebral cortex after ischemia-reperfusion injury.

Chen YH, Chiang YH, Ma HI - J Clin Neurol (2014)

Bottom Line: This study elucidated the mechanisms underlying ischemia-induced brain injury that results in neurological sequelae.The level of 78-kDa glucose-regulated protein precursor on the lesioned side of the cerebral cortex was found to be high initially, but then down-regulated by 24 h after the induction of ischemia-reperfusion injury.These findings provide insights into the mechanisms underlying the neurodegenerative events that occur following cerebral ischemia.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurological Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan, ROC.

ABSTRACT

Background and purpose: Hypoxia, or ischemia, is a common cause of neurological deficits in the elderly. This study elucidated the mechanisms underlying ischemia-induced brain injury that results in neurological sequelae.

Methods: Cerebral ischemia was induced in male Sprague-Dawley rats by transient ligation of the left carotid artery followed by 60 min of hypoxia. A two-dimensional differential proteome analysis was performed using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry to compare changes in protein expression on the lesioned side of the cortex relative to that on the contralateral side at 0, 6, and 24 h after ischemia.

Results: The expressions of the following five proteins were up-regulated in the ipsilateral cortex at 24 h after ischemia-reperfusion injury compared to the contralateral (i.e., control) side: aconitase 2, neurotensin-related peptide, hypothetical protein XP-212759, 60-kDa heat-shock protein, and aldolase A. The expression of one protein, dynamin-1, was up-regulated only at the 6-h time point. The level of 78-kDa glucose-regulated protein precursor on the lesioned side of the cerebral cortex was found to be high initially, but then down-regulated by 24 h after the induction of ischemia-reperfusion injury. The expressions of several metabolic enzymes and translational factors were also perturbed soon after brain ischemia.

Conclusions: These findings provide insights into the mechanisms underlying the neurodegenerative events that occur following cerebral ischemia.

No MeSH data available.


Related in: MedlinePlus

Temporal proteomes of ischemia-reperfusion injury in the cerebrocortex: two types of stain (silver stain and SYPRO Ruby) were used on 2-D gels at various time points after ischemia-reperfusion injury.
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Figure 3: Temporal proteomes of ischemia-reperfusion injury in the cerebrocortex: two types of stain (silver stain and SYPRO Ruby) were used on 2-D gels at various time points after ischemia-reperfusion injury.

Mentions: In addition to silver staining, SYPRO Ruby protein gel staining was performed to investigate the protein profile in the rat brain during the reperfusion stage following tMCAO. The 2-DE maps of sham-operated animals were compared at 6 and 24 h after tMCAO (Fig. 3). About 400 spots were found and histograms of each spot at different time points (0, 6, and 24 h following tMCAO) were compared (Fig. 4). The silver stain did not reveal significant up- or down-regulation of the protein profile, but the SYPRO Ruby stain revealed 8 spots of down-regulation and 39 spots of up-regulation at 24 h after reperfusion. On the other hand, 11 spots were up-regulated at 6 h and then returned to noninjury levels by 24 h. After in-gel digestion, the proteins were eluted from the spots and fingerprint patterns of peptides were prescribed by MALDI-TOF analysis. The proteins were identified after Mascot search comparison (Fig. 5). Of these, only seven were successfully identified by in-gel digestion by MALDI-TOF analysis, with a protein sequence coverage of 17-31% and top scores ranging from 62 to 111 (Table 1). Five spots were identified as aconitase 2, neurotensin-related peptide (NRP), hypothetical protein XP-212759, 60-kDa heat-shock protein (HSP60), and aldolase A. The expression levels of these proteins were up-regulated by 1.43-5 times in comparison to their baseline expressions in the histograms. The observed molecular masses differed from the theoretical values by no more than 5-6%, and the observed isoelectric point values were also in close agreement with the theoretical values.


Analysis of spatial and temporal protein expression in the cerebral cortex after ischemia-reperfusion injury.

Chen YH, Chiang YH, Ma HI - J Clin Neurol (2014)

Temporal proteomes of ischemia-reperfusion injury in the cerebrocortex: two types of stain (silver stain and SYPRO Ruby) were used on 2-D gels at various time points after ischemia-reperfusion injury.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4017024&req=5

Figure 3: Temporal proteomes of ischemia-reperfusion injury in the cerebrocortex: two types of stain (silver stain and SYPRO Ruby) were used on 2-D gels at various time points after ischemia-reperfusion injury.
Mentions: In addition to silver staining, SYPRO Ruby protein gel staining was performed to investigate the protein profile in the rat brain during the reperfusion stage following tMCAO. The 2-DE maps of sham-operated animals were compared at 6 and 24 h after tMCAO (Fig. 3). About 400 spots were found and histograms of each spot at different time points (0, 6, and 24 h following tMCAO) were compared (Fig. 4). The silver stain did not reveal significant up- or down-regulation of the protein profile, but the SYPRO Ruby stain revealed 8 spots of down-regulation and 39 spots of up-regulation at 24 h after reperfusion. On the other hand, 11 spots were up-regulated at 6 h and then returned to noninjury levels by 24 h. After in-gel digestion, the proteins were eluted from the spots and fingerprint patterns of peptides were prescribed by MALDI-TOF analysis. The proteins were identified after Mascot search comparison (Fig. 5). Of these, only seven were successfully identified by in-gel digestion by MALDI-TOF analysis, with a protein sequence coverage of 17-31% and top scores ranging from 62 to 111 (Table 1). Five spots were identified as aconitase 2, neurotensin-related peptide (NRP), hypothetical protein XP-212759, 60-kDa heat-shock protein (HSP60), and aldolase A. The expression levels of these proteins were up-regulated by 1.43-5 times in comparison to their baseline expressions in the histograms. The observed molecular masses differed from the theoretical values by no more than 5-6%, and the observed isoelectric point values were also in close agreement with the theoretical values.

Bottom Line: This study elucidated the mechanisms underlying ischemia-induced brain injury that results in neurological sequelae.The level of 78-kDa glucose-regulated protein precursor on the lesioned side of the cerebral cortex was found to be high initially, but then down-regulated by 24 h after the induction of ischemia-reperfusion injury.These findings provide insights into the mechanisms underlying the neurodegenerative events that occur following cerebral ischemia.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurological Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan, ROC.

ABSTRACT

Background and purpose: Hypoxia, or ischemia, is a common cause of neurological deficits in the elderly. This study elucidated the mechanisms underlying ischemia-induced brain injury that results in neurological sequelae.

Methods: Cerebral ischemia was induced in male Sprague-Dawley rats by transient ligation of the left carotid artery followed by 60 min of hypoxia. A two-dimensional differential proteome analysis was performed using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry to compare changes in protein expression on the lesioned side of the cortex relative to that on the contralateral side at 0, 6, and 24 h after ischemia.

Results: The expressions of the following five proteins were up-regulated in the ipsilateral cortex at 24 h after ischemia-reperfusion injury compared to the contralateral (i.e., control) side: aconitase 2, neurotensin-related peptide, hypothetical protein XP-212759, 60-kDa heat-shock protein, and aldolase A. The expression of one protein, dynamin-1, was up-regulated only at the 6-h time point. The level of 78-kDa glucose-regulated protein precursor on the lesioned side of the cerebral cortex was found to be high initially, but then down-regulated by 24 h after the induction of ischemia-reperfusion injury. The expressions of several metabolic enzymes and translational factors were also perturbed soon after brain ischemia.

Conclusions: These findings provide insights into the mechanisms underlying the neurodegenerative events that occur following cerebral ischemia.

No MeSH data available.


Related in: MedlinePlus