{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T19:58:06Z","timestamp":1790366286307,"version":"4.1.0"},"update-to":[{"DOI":"10.1371\/journal.ppat.1012733","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2025,6,25]],"date-time":"2025-06-25T00:00:00Z","timestamp":1750809600000}}],"reference-count":143,"publisher":"Public Library of Science (PLoS)","issue":"6","license":[{"start":{"date-parts":[[2025,6,16]],"date-time":"2025-06-16T00:00:00Z","timestamp":1750032000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH","award":["NS137603 and AI158417"],"award-info":[{"award-number":["NS137603 and AI158417"]}]},{"name":"Graduate Division at the University of California, Riverside"},{"name":"Division of Biomedical Sciences, UCR"}],"content-domain":{"domain":["www.plospathogens.org"],"crossmark-restriction":false},"short-container-title":["PLoS Pathog"],"abstract":"<jats:p><jats:italic>Toxoplasma gondii (T. gondii),<\/jats:italic> a prolific protozoan parasite, forms cysts within neurons of the central nervous system that maintain infection for the lifetime of the host. Astrocytes are fundamental to neuronal health by providing nutrients and structural support and help regulate neurotransmitters by continuous communication with neurons. It is not yet known how infection and the presence of intracellular cysts, disrupts the crucial relationship between these cells. Extracellular vesicles (EVs) function in intracellular communication and can contain proteins, lipids, DNA, miRNA, and other RNA subtypes. EVs are produced by all cells and play an important role in neuronal-astrocyte interactions, including the regulation of glutamate receptors on astrocytes. Previous work has demonstrated that Toxoplasma infection reduces astrocytic expression of the primary glutamate transporter, GLT-1. Here we tested if cyst infection of neurons alters the production and content of EVs. EVs were isolated from uninfected and infected primary murine cortical neurons and their size, concentration, and characterization were confirmed with nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), ELISA, western blot, liquid chromatography (LC)-mass spectrometry (MS)\/MS, and microRNA sequencing. Analysis reveals that infection of neurons reduced neuronal production of EVs and altered their protein and miRNA content. In addition to changes in host protein content, EVs from infected neurons contained the Toxoplasma proteins GRA1, GRA2, GRA7, MAG1 and MAG2. Following incubation of neuronal EVs with primary astrocytes, GRA7 protein could be observed within intracellular EVs and the nuclei of GRA7<jats:sup>\u2009+<\/jats:sup>\u2009EV-containing cells. EVs from infected neurons altered gene expression of astrocytes resulting in an increase in pro-inflammatory transcriptional signatures, along with a downregulation of GLT-1 protein expression with similar transcriptional changes found in astrocytes <jats:italic>in vivo<\/jats:italic>. These results demonstrate the ability of a parasitic infection in the brain to alter EV production and the fundamental communication between neurons and astrocytes.<\/jats:p>","DOI":"10.1371\/journal.ppat.1012733","type":"journal-article","created":{"date-parts":[[2025,6,16]],"date-time":"2025-06-16T17:43:38Z","timestamp":1750095818000},"page":"e1012733","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":7,"title":["Toxoplasma gondii infection of neurons alters the production and content of extracellular vesicles directing astrocyte phenotype and contributing to the loss of GLT-1 in the infected brain"],"prefix":"10.1371","volume":"21","author":[{"given":"Emily Z.","family":"Tabaie","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziting","family":"Gao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nala","family":"Kachour","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arzu","family":"Ulu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stacey","family":"Gomez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zoe A.","family":"Figueroa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kristina V.","family":"Bergersen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenwan","family":"Zhong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6054-9981","authenticated-orcid":true,"given":"Emma H.","family":"Wilson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2025,6,16]]},"reference":[{"issue":"1","key":"ppat.1012733.ref001","first-page":"48","article-title":"Toxoplasma and toxoplasmosis","volume":"142","author":"J el-On","year":"2003","journal-title":"Harefuah"},{"key":"ppat.1012733.ref002","doi-asserted-by":"crossref","first-page":"788303","DOI":"10.3389\/fcimb.2021.788303","article-title":"Depletion of intracellular glutamine pools triggers Toxoplasma gondii stage conversion in human glutamatergic neurons","volume":"11","author":"H Bando","year":"2022","journal-title":"Front Cell Infect Microbiol"},{"issue":"7","key":"ppat.1012733.ref003","doi-asserted-by":"crossref","DOI":"10.1371\/journal.ppat.1006351","article-title":"Toxoplasma gondii: Entry, association, and physiological influence on the central nervous system. 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