Quantitative mass spectrometric analysis of the mouse cerebral cortex after ischemic stroke

Abstract

Ischemic strokes result in the death of brain tissue and a wave of downstream effects, often leading to lifelong disabilities or death. However, the underlying mechanisms of ischemic damage and repair systems remain largely unknown. In order to better understand these mechanisms, TMT-isobaric mass tagging and mass spectrometry were conducted on brain cortex extracts from mice subjected to one hour of middle cerebral artery occlusion (MCAO) and after one hour of reperfusion. In total, 2,690 proteins were identified and quantified, out of which 65% of the top 5% of up-and down-regulated proteins were found to be significant (p < 0.05). Network-based gene ontology analysis was then utilized to cluster all identified proteins by protein functional groups and cellular roles. Although three different cellular functions were identified-organelle outer membrane proteins, cytosolic ribosome proteins, and spliceosome complex proteins-several functional domains were found to be common. Of these, organelle outer membrane proteins were downregulated whereas cytosolic ribosome and spliceosome complex proteins were upregulated, indicating that major molecular events post-stroke were translation-associated and subsequent signaling pathways (e.g., poly (ADP-ribose) (PAR) dependent cell death). By approaching stroke analyses via TMT-isobaric mass tagging, the work herein presents a grand scope of protein-based molecular mechanisms involved with ischemic stroke recovery.

Proteomic Characterization of the Dynamics of Ischemic Stroke in Mice

“…Several proteomic analyses of AIS using MCAO models − have been reported. They were conducted on transient MCAO mouse or rat models and generally focused on the acute phase poststroke.…”

Identifying Key Biomarkers and Immune Infiltration in Female Patients with Ischemic Stroke Based on Weighted Gene Co-Expression Network Analysis

“…Ribosomal proteins (RPs) play an important role in the regulation of gene expression and protein synthesis [19]. Elsewhere, an experimental study found that cytoplasmic ribosomes play a role in functional recovery after ischemic stroke [20]. In the present study, the GO and KEGG results revealed that immune response and ribosomes are critical links in the mechanism of female patients with ischemic stroke.…”

ERK1/2 Activity Is Critical for the Outcome of Ischemic Stroke

“…To evaluate the function of the regulated proteins in the ipsilateral hemisphere and their impact on stroke outcome, we assigned the significantly regulated proteins to gene ontology functional categories (GO-term: biological process) using a cut-off for a 2-fold upregulation and for a 0.5-fold downregulation compared to wt bgi. These results show that the regulated proteins were found within the signaling processes commonly involved after ischemic brain injury, such as cell proliferation, cell differentiation, cell growth, cell death, translation, transcription, synaptic plasticity, signal transduction, metabolism, protein transport, vesicle transport, proteolysis, cell adhesion, membrane potential, immune response/inflammation, cytoskeleton organization, and regulation of hydrogen peroxide ( Tables S6–S8 ) [ 33 , 34 , 35 ]. As these processes were affected in all genotypes, the analyses did not allow the identification of a main mechanism for the respective genotypes.…”