{"database":"Pride","file_versions":[{"headers":{"Content-Type":["application/json"]},"body":{"files":{"Raw":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD055523/2311_YY_06.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD055523/2311_YY_04.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD055523/2311_YY_18.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD055523/2311_YY_12.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD055523/2311_YY_24.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD055523/2311_YY_10.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD055523/2311_YY_16.raw","ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD055523/2311_YY_22.raw"],"Other":["ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/06/PXD055523/2311_YY_compartment.zip"]},"type":"primary"},"statusCode":"OK","statusCodeValue":200}],"scores":null,"additional":{"labhead_mail":["richard.wilson@utas.edu.au"],"submitter":["Richard Wilson"],"technology_type":["Mass Spectrometry","Data-independent acquisition"],"software":[""],"submitter_keywords":["Astrocytes","Neuropathology","Proteomics","Microfluidics"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD055523"],"tissue":["Cell Culture"],"sample_protocol":["Proteins were harvested from microfluidic chamber compartments in devices at 7 DIV of neuron/astrocytes co-culture. Cells was washed three times with cold PBS then lysed in lysis buffer containing urea (7M, Sigma), thiourea (2M, Sigma), Tris-HCl (30mM, Sigma), dithiothreitol (DTT, 10 mM, Sigma) and EDTA-free protease inhibitors (Roche). Cell lysates were sonicated for three cycles of 15 second pulse with 5 second intervals on ice. Cell lysates were mixed gently on a rotary suspension mixer at 4°C for 16 h and centrifuged at 16,000 g for 30 min before supernatant was collected in protein lobind tubes. Protein concentration was quantified using by Pierce 660 nm Protein Assay (Thermo Fisher Scientific) according to the manufacturer's instructions. Proteins (20 µg/sample) were sequentially reduced overnight at 4oC using DTT (10 mM, Sigma) and alkylation with iodoacetamide (IAA, 50 mM, Sigma) for 2 hours at room temperature in the dark. The samples underwent single-pot, solid-phase-enhanced sample-preparation (SP3), followed by digestion with 1.2 µg proteomic-grade trypsin/rLysC (Promega) overnight at 37°C."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["The DIA MS raw files were processed using Spectronaut software (version 18, Biognosys AB). A project-specific library comprising 6,308 protein groups was generated using the Pulsar search engine to search the DIA MS1 spectra against the Rattus norvegicus (Rat), UniProt reference proteome UP000002494 concatenated with common contaminants (comprising 22,898 entries, downloaded in November 2023). With the exception that single-hit proteins were excluded, default (Biognosys factory) settings were used for both spectral library generation and DIA data extraction. For library generation, these included N-terminal acetylation and methionine oxidation as variable modifications and cysteine carbamidomethylation as a fixed modification, up to two missed cleavages allowed and peptide, protein and peptide spectrum match thresholds set to 0.01. Mass tolerances were based on first-pass calibration and extensive calibration, for the calibration and main searches, respectively, with correction factors set to 1 at the MS1 and MS2 levels. Targeted searching of the library based on ion current extraction deployed dynamic retention time alignment with a correction factor of 1"],"omics_type":["Proteomics"],"labhead":["Dr Richard Wilson"],"instrument_platform":[""],"labhead_affiliation":["Proteomics facility, Central Science Laboratory, University of Tasmania"],"submission_type":["PARTIAL"],"species":["Rattus Norvegicus (rat)"],"submitter_mail":["richard.wilson@utas.edu.au"],"publication":["42129138 Yap YC, Musgrove RE, Breadmore MC, Guijt RM, Wilson R, Wertheimer G, King AE, Dickson TC. Microfluidic co-culture system for synaptically segregated neural networks to explore astrocyte-driven neural pathology. Microsyst Nanoeng. 2026 12(1):181 10.1038/s41378-026-01187-3"],"submitter_affiliation":["University of Tasmania"],"submitter_country":["Australia"],"pubmed_abstract":["Investigating astrocyte-neuron communication in the absence of neuron-to-neuron signalling is challenging using traditional culture systems due to the complexity of synaptic networks. To address this, we designed a three-compartment microfluidic co-culture device that fluidically isolates two neuronal populations while permitting astrocyte growth throughout. This design enables assessment of astrocyte-specific contributions to neuropathology between synaptically segregated neurons. The device incorporates ten microchannel banks forming maze-like structures that restrict neurite extension and fluid exchange, while allowing an astrocyte monolayer to infiltrate all compartments. Using this platform, we exposed one neuron-astrocyte population to the excitotoxin kainic acid (KA) and observed neurite degeneration in the adjacent, fluidically isolated neurons connected only via astrocytes. Pre-treatment of astrocytes with the membrane-permeable chelator BAPTA-AM markedly attenuated this effect, implicating calcium in astrocyte-mediated excitotoxicity. This microfluidic system provides a controllable in vitro model of neuron-astrocyte networks, enabling directional connectivity and mechanistic studies of circuit behaviour. Our findings highlight the utility of this platform for exploring intercellular signalling pathways relevant to neurodegenerative disease."],"pubmed_title":["Microfluidic co-culture system for synaptically segregated neural networks to explore astrocyte-driven neural pathology."],"pubmed_authors":["Yap Yiing C YC, Musgrove Ruth E RE, Breadmore Michael C MC, Guijt Rosanne M RM, Wilson Richard R, Wertheimer Graeme G, King Anna E AE, Dickson Tracey C TC"],"additional_accession":[]},"is_claimable":false,"name":"A microfluidic co-culture platform to generate synaptically separated networks that uncover the role of astrocytes in the initiation and progression of neuron pathology","description":"Communication between astrocytes and neurons plays a pivotal role in the development, maintenance and dysfunction of cellular networks within the central nervous system. Further understanding the contribution of astrocytes to the initiation and progression of neuropathology has, however, been hindered by the complexity of cell networks in traditional in vivo and ex vivo systems. We have designed and constructed a novel three-compartment microfluidic cell culture device that enables us to overcome these limitations and uncouple the roles of astrocytes and neurons in the transfer of pathology through complex cell networks. Our microfluidic device integrates a novel maze-like structure that prevents synaptic connectivity between two fluidically isolated neuron populations, while allowing astrocyte infiltration and growth throughout. We use primary neuron/astrocyte co-cultures, proteomic analysis and immunocytochemistry to validate the application of this device. Using this device, we identify and describe a novel calcium-dependent role for astrocytes in the transfer of excitotoxic pathology between the segregated neuron 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