{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,23]],"date-time":"2026-06-23T12:49:43Z","timestamp":1782218983354,"version":"3.54.5"},"reference-count":53,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2023,2,21]],"date-time":"2023-02-21T00:00:00Z","timestamp":1676937600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100016887","name":"Motor Neuron Disease Research Australia Innovator","doi-asserted-by":"publisher","award":["IG1910"],"award-info":[{"award-number":["IG1910"]}],"id":[{"id":"10.13039\/100016887","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100016887","name":"Motor Neuron Disease Research Australia Innovator","doi-asserted-by":"publisher","award":["IG2221"],"award-info":[{"award-number":["IG2221"]}],"id":[{"id":"10.13039\/100016887","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100016887","name":"Motor Neuron Disease Research Australia Innovator","doi-asserted-by":"publisher","award":["IG2308"],"award-info":[{"award-number":["IG2308"]}],"id":[{"id":"10.13039\/100016887","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100016887","name":"Motor Neuron Disease Research Australia Innovator","doi-asserted-by":"publisher","award":["DP210103469"],"award-info":[{"award-number":["DP210103469"]}],"id":[{"id":"10.13039\/100016887","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Australian Research Council Discovery","award":["IG1910"],"award-info":[{"award-number":["IG1910"]}]},{"name":"Australian Research Council Discovery","award":["IG2221"],"award-info":[{"award-number":["IG2221"]}]},{"name":"Australian Research Council Discovery","award":["IG2308"],"award-info":[{"award-number":["IG2308"]}]},{"name":"Australian Research Council Discovery","award":["DP210103469"],"award-info":[{"award-number":["DP210103469"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Proteomics offers vast potential for studying the molecular regulation of the human brain. Formalin fixation is a common method for preserving human tissue; however, it presents challenges for proteomic analysis. In this study, we compared the efficiency of two different protein-extraction buffers on three post-mortem, formalin-fixed human brains. Equal amounts of extracted proteins were subjected to in-gel tryptic digestion and LC-MS\/MS. Protein, peptide sequence, and peptide group identifications; protein abundance; and gene ontology pathways were analyzed. Protein extraction was superior using lysis buffer containing tris(hydroxymethyl)aminomethane hydrochloride, sodium dodecyl sulfate, sodium deoxycholate, and Triton X-100 (TrisHCl, SDS, SDC, Triton X-100), which was then used for inter-regional analysis. Pre-frontal, motor, temporal, and occipital cortex tissues were analyzed by label free quantification (LFQ) proteomics, Ingenuity Pathway Analysis and PANTHERdb. Inter-regional analysis revealed differential enrichment of proteins. We found similarly activated cellular signaling pathways in different brain regions, suggesting commonalities in the molecular regulation of neuroanatomically-linked brain functions. Overall, we developed an optimized, robust, and efficient method for protein extraction from formalin-fixed human brain tissue for in-depth LFQ proteomics. We also demonstrate herein that this method is suitable for rapid and routine analysis to uncover molecular signaling pathways in the human brain.<\/jats:p>","DOI":"10.3390\/ijms24054283","type":"journal-article","created":{"date-parts":[[2023,2,22]],"date-time":"2023-02-22T03:32:17Z","timestamp":1677036737000},"page":"4283","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Inter-Regional Proteomic Profiling of the Human Brain Using an Optimized Protein Extraction Method from Formalin-Fixed Tissue to Identify Signaling Pathways"],"prefix":"10.3390","volume":"24","author":[{"given":"Jennilee M.","family":"Davidson","sequence":"first","affiliation":[{"name":"Centre for Motor Neuron Disease Research, Macquarie Medical School, Faculty of Medicine, Health and Human Sciences, Macquarie University, Level 1, 75 Talavera Road, Sydney, NSW 2109, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephanie L.","family":"Rayner","sequence":"additional","affiliation":[{"name":"Centre for Motor Neuron Disease Research, Macquarie Medical School, Faculty of Medicine, Health and Human Sciences, Macquarie University, Level 1, 75 Talavera Road, Sydney, NSW 2109, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sidong","family":"Liu","sequence":"additional","affiliation":[{"name":"Centre for Health Informatics, Faculty of Medicine, Health and Human Sciences, Macquarie University, 75 Talavera Road, Sydney, NSW 2109, Australia"},{"name":"Computational NeuroSurgery (CNS) Lab, Macquarie Medical School, Faculty of Medicine, Health and Human Sciences, Macquarie University, Level 1, 75 Talavera Road, Sydney, NSW 2109, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Flora","family":"Cheng","sequence":"additional","affiliation":[{"name":"Centre for Motor Neuron Disease Research, Macquarie Medical School, Faculty of Medicine, Health and Human Sciences, Macquarie University, Level 1, 75 Talavera Road, Sydney, NSW 2109, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antonio","family":"Di Ieva","sequence":"additional","affiliation":[{"name":"Computational NeuroSurgery (CNS) Lab, Macquarie Medical School, Faculty of Medicine, Health and Human Sciences, Macquarie University, Level 1, 75 Talavera Road, Sydney, NSW 2109, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roger S.","family":"Chung","sequence":"additional","affiliation":[{"name":"Centre for Motor Neuron Disease Research, Macquarie Medical School, Faculty of Medicine, Health and Human Sciences, Macquarie University, Level 1, 75 Talavera Road, Sydney, NSW 2109, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5156-0567","authenticated-orcid":false,"given":"Albert","family":"Lee","sequence":"additional","affiliation":[{"name":"Centre for Motor Neuron Disease Research, Macquarie Medical School, Faculty of Medicine, Health and Human Sciences, Macquarie University, Level 1, 75 Talavera Road, Sydney, NSW 2109, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,2,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1038\/nature18933","article-title":"A multi-modal parcellation of human cerebral cortex","volume":"536","author":"Glasser","year":"2016","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"4933","DOI":"10.1038\/s41467-022-32595-4","article-title":"Population-based tract-to-region connectome of the human brain and its hierarchical topology","volume":"13","author":"Yeh","year":"2022","journal-title":"Nat. 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