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<p>Stroke remains a major global health challenge due to its high mortality and significant socioeconomic burden. Despite advances in clinical management, effective diagnostic tools and therapeutic strategies remain limited. This study aimed to identify and expand the repertoire of biomarkers of damage and repair that could serve as potential diagnostic and prognostic tools across post-stroke phases. Twenty-three male wild-type mice were assigned according to three longitudinal time points to control pre-stroke, 24-hour acute, and 35-day chronic post-stroke groups. Ischemic injury was induced via a 30-minute middle cerebral artery occlusion Koizumi method. Magnetic resonance imaging and neurological scoring were used to assess lesion size and functional deficit acutely, as well as structural and functional recovery during the chronic phase. Proteomic profiling of the ipsilateral and contralateral cortices was performed using data-independent acquisition (DIA)-based MS method. Statistical analysis revealed 74 differentially expressed proteins showing significant temporal changes in expression, which were classified into four temporal expression clusters: <i>acutely and chronically upregulated</i>,<i> acutely upregulated and chronically downregulated</i>, <i>acutely downregulated and chronically upregulated</i>, and <i>acutely and chronically downregulated</i>. Gene ontology analysis identified 47 affected biological processes, including synaptic signaling, immune response, cell-cell communication, cytoskeletal organization, and proliferation. Thirteen proteins previously not associated with stroke pathophysiology were identified, including 10 from the ipsilateral cortex (<i>Dbi</i>, <i>Cpne3</i>, <i>Dnm2</i>, <i>Eef1a1</i>, <i>Taldo1</i>, <i>Pgls</i>, <i>Gnb5</i>, <i>Phf24</i>, <i>Ctsz</i>, <i>Capg</i>) and 3 from the contralateral cortex (<i>Agpat3</i>, <i>Cacng8</i>, <i>Endod</i>). The identified biomarkers provide novel molecular insights into post-stroke energy metabolism, neuroinflammation, and cellular remodeling, highlighting potential targets for further intervention.&#xa0;</p> Graphical Abstract <p>Proteomic analysis of ipsilateral and contralateral mouse cortices post-stroke identified 13 previously unreported proteins, revealing distinct temporal expression patterns linked to tissue damage and repair. These findings highlight potential biomarkers with diagnostic, prognostic, or predictive value for ischemic stroke.</p> <p></p>

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Proteomic Analysis of Mouse Cerebral Cortex Following Experimental Ischemic Stroke: Identifying Novel Biomarkers of Damage and Repair

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年12月4日

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  • Dominik Hamer,
  • Ana Butorac,
  • Daniela Petrinec,
  • Monika Berecki,
  • Vera M. Mendes,
  • Bruno Manadas,
  • Vanja Kelava,
  • Branimir K. Hackenberger,
  • Anton Glasnović,
  • Marija Lovrić,
  • Srećko Gajović,
  • Marina Dobrivojević Radmilović

摘要

Stroke remains a major global health challenge due to its high mortality and significant socioeconomic burden. Despite advances in clinical management, effective diagnostic tools and therapeutic strategies remain limited. This study aimed to identify and expand the repertoire of biomarkers of damage and repair that could serve as potential diagnostic and prognostic tools across post-stroke phases. Twenty-three male wild-type mice were assigned according to three longitudinal time points to control pre-stroke, 24-hour acute, and 35-day chronic post-stroke groups. Ischemic injury was induced via a 30-minute middle cerebral artery occlusion Koizumi method. Magnetic resonance imaging and neurological scoring were used to assess lesion size and functional deficit acutely, as well as structural and functional recovery during the chronic phase. Proteomic profiling of the ipsilateral and contralateral cortices was performed using data-independent acquisition (DIA)-based MS method. Statistical analysis revealed 74 differentially expressed proteins showing significant temporal changes in expression, which were classified into four temporal expression clusters: acutely and chronically upregulated, acutely upregulated and chronically downregulated, acutely downregulated and chronically upregulated, and acutely and chronically downregulated. Gene ontology analysis identified 47 affected biological processes, including synaptic signaling, immune response, cell-cell communication, cytoskeletal organization, and proliferation. Thirteen proteins previously not associated with stroke pathophysiology were identified, including 10 from the ipsilateral cortex (Dbi, Cpne3, Dnm2, Eef1a1, Taldo1, Pgls, Gnb5, Phf24, Ctsz, Capg) and 3 from the contralateral cortex (Agpat3, Cacng8, Endod). The identified biomarkers provide novel molecular insights into post-stroke energy metabolism, neuroinflammation, and cellular remodeling, highlighting potential targets for further intervention. 

Graphical Abstract

Proteomic analysis of ipsilateral and contralateral mouse cortices post-stroke identified 13 previously unreported proteins, revealing distinct temporal expression patterns linked to tissue damage and repair. These findings highlight potential biomarkers with diagnostic, prognostic, or predictive value for ischemic stroke.