Effects of Cocaine on Human Glial-Derived Extracellular Vesicles

Abstract

BackgroundMicroglia are important myeloid cells present in the brain parenchyma that serve a surveillance function in the central nervous system. Microglial cell activation results in neuroinflammation that, when prolonged, can disrupt immune homeostasis and neurogenesis. Activated microglia-derived extracellular vesicles (EVs) may be involved in the propagation of inflammatory responses and modulation of cell-to-cell communication. However, a complete understanding of how EVs are regulated by drugs of abuse, such as cocaine, is still lacking.FindingsCocaine exposure reduced human microglial cell (HMC3) viability, decreased expression of CD63 and dectin-1 in HMC3-derived EVs, and increased expression of the apoptotic marker histone H2A.x in HMC3-derived EVs.ConclusionCocaine impacts HMC3 cell viability and specific EV protein expression, which could disrupt cellular signaling and cell-to-cell communication.

Human Adenovirus Serotype 3 Infection Modulates the Biogenesis and Composition of Lung Cell-Derived Extracellular Vesicles

“…EVs isolated from untreated cells and infected cells at MOI 300 were morphologically similar ( Figure 2(a) ). A549-derived EVs were in the size range (30-150 nm) of the previously documented size array of exosomes and were consistent with previous studies (Figures 2(b) and 2(c) ) [ 42 ]. Results showed a reduction in EVs derived from infected cells (particle per mL) relative to the uninfected control EVs, although sizes were not significantly altered (Figures 2(d) and 2(e) ).…”

“…Plates were incubated for 24, 48, and 72 hours (h), and each time point was evaluated as an independent experiment. Infected cell supernatant was collected and stored for EV isolation at the end of incubation period [ 42 ].…”

“…The color formation was read at 570 nm and a reference wavelength of 630 nm and stored at 4°C. All samples were evaluated in triplicate [ 24 , 42 ].…”

“…EVs were isolated and purified from DMEM/F12 exosome-free cell culture media. In brief, EVs were isolated as previously described [ 24 , 25 , 42 , 43 ]. The media was collected after infection and spun down at 1,300 revolutions per minute (rpm) at 4°C for 10 min, using a Sorvall RT 6000 refrigerated centrifuge.…”

“…Approximately 500 μ L of purified EVs was collected below the meniscus of the centrifuge tube. Collected EVs were quantified using the Bradford–Lowry quantitation method [ 24 , 42 , 44 ].…”