Detection, visualization and quantification of protein complexes in human Alzheimer’s disease brains using proximity ligation assay

Autor: Wilber Romero-Fernandez, Cristian Carvajal-Tapia, Alex Prusky, Ketaki A. Katdare, Emmeline Wang, Alena Shostak, Lissa Ventura-Antunes, Hannah J. Harmsen, Ethan S. Lippmann, Kjell Fuxe, Jason A. MacGurn, Dasiel O. Borroto-Escuela, Matthew S. Schrag
Jazyk: angličtina
Rok vydání: 2023
Předmět:
Zdroj: Scientific Reports, Vol 13, Iss 1, Pp 1-13 (2023)
Druh dokumentu: article
ISSN: 2045-2322
DOI: 10.1038/s41598-023-38000-4
Popis: Abstract Examination of healthy and diseased human brain is essential to translational neuroscience. Protein–protein interactions play a pivotal role in physiological and pathological processes, but their detection is difficult, especially in aged and fixed human brain tissue. We used the in-situ proximity ligation assay (PLA) to broaden the range of molecular interactions assessable in-situ in the human neuropathology. We adapted fluorescent in-situ PLA to detect ubiquitin-modified proteins in human brains with Alzheimer’s disease (AD), including approaches for the management of autofluorescence and quantification using a high-content image analysis system. We confirmed that phosphorylated microtubule-associated protein tau (Serine202, Threonine205) aggregates were modified by ubiquitin and that phospho-tau-ubiquitin complexes were increased in hippocampal and frontal cortex regions in AD compared to non-AD brains. Overall, we refined PLA for use in human neuropathology, which has revealed a profound change in the distribution of ubiquitin in AD brain and its association with characteristic tau pathologies.
Databáze: Directory of Open Access Journals
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