Journal: Journal of Neurovirology

Article Title: SARS-CoV and SARS-CoV-2 display limited neuronal infection and lack the ability to transmit within synaptically connected axons in stem cell–derived human neurons

doi: 10.1007/s13365-023-01187-3

Figure Lengend Snippet: Sarbecovirus infection of ex vivo model of interconnected human neural network to assess trans-synaptic transmission. A Stem cell–derived NPCs were seeded in both panels of microfluidic device separated by microchannels which allows only the neurites to pass through. After seeding, NPCs were differentiated for 20 (RABV) to 21 days (sarbecoviruses) to generate neurons in both panels interconnected by neurites through the microchannels. The neuronal network is identified by TUJ1 staining (red) and F-actin stain to visualise the neural architecture (magenta). The ex vivo models were infected MOI 1 with RABV for 48 h, which served as a positive control for trans-synaptic transmission ( B ) or different strains of sarbecoviruses ( C, D, E ) at for 24 h. To prevent passive diffusion of virus from the infected panel into the non-infected panel, a higher volume of media is maintained in the non-infected panel. After 24 h of infection, neural cultures were fixed, stained, and imaged in the microfluidic device. Tile images were taken with 20 × objective with Z-stacks and stitched together. Image B shows trans-synaptic spread of RABV from the infected to non-infected panel identified by staining with a rabies anti-nucleoprotein antibody (green). Images C, D, and E show SARS and SARS-CoV-2 infections stained with anti-SARS2-S1 antibody (green) in a subset of neurons in the infected panel (boxed area) but viral antigen was not observed in the non-infected panel

Article Snippet: Xona microfluidic devices (Xona Microfluidics, Cat#SND450) were sterilised and plasma bonded to glass coverslips (24 × 40 mm; Menzel Glaser) using a plasma cleaner (PDC-32G-2, Harrick Plasma).

Techniques: Infection, Ex Vivo, Transmission Assay, Derivative Assay, Staining, Positive Control, Diffusion-based Assay, Virus

Journal: Journal of Extracellular Vesicles

Article Title: Small extracellular vesicles ameliorate peripheral neuropathy and enhance chemotherapy of oxaliplatin on ovarian cancer

doi: 10.1002/jev2.12073

Figure Lengend Snippet: CEC‐sEVs promote axonal growth of DRG neurons with the presence of oxaliplatin. [Representative confocal microscopic images (a) show CECs are CD31 (green, CD31) and ZO1 positive (red, ZO‐1). Characterization of CEC‐sEVs by TEM (b), NTA (c) and Western blots (d), respectively. Schematic figure of the standard microfluidic device (SND150) along with an immunofluorescent image captured in the box area (e) shows DRG neurons grown in the cell body compartment (DRG soma) and their axons in 150 μm long microgrooves and in the axonal compartment (Axon). Representative time‐lapse microscopic images (f) of growth cone extension within 60 min and corresponding quantitative data of growth cone extension during a 24 h period (g), respectively, under control (con), CEC‐sEVs (sEV), oxaliplatin (oxa) and CEC‐sEVs in combination with oxaliplatin (sEV+oxa) conditions. Yellow and red arrows in panel G indicate the start (0′) and end positions (60′), respectively. One‐way ANOVA with Tukey's multiple comparisons test was used. *** P < 0.001 vs. control. N indicates the number of axonal growth cones. In panel D, NCM = the particles isolated from non‐conditioned medium, sEVs = CEC‐sEVs, Ept = the intentionally empty lanes, Cell = CEC lysate, K = the molecular weight Kda. Error bars indicate the standard error of the mean (SEM)]

Article Snippet: To separate axons from neuronal soma and examine the effects of oxaliplatin/CEC‐sEVs on axons of DRG neurons, a microfluidic device (Standard Neuron Device, Cat# SND150, Xona Microfluidics, Temecula, CA) was employed (Figure ) (Jia et al., ; Wang et al., ; Zhang et al., ).

Techniques: Western Blot, Control, Isolation, Molecular Weight

Journal: eLife

Article Title: Axonal T3 uptake and transport can trigger thyroid hormone signaling in the brain

doi: 10.7554/eLife.82683

Figure Lengend Snippet: ( A ) Microfluidic device showing the MC-CS in blue and the MC-AS in orange. ( B ). Typical neuronal growth at DIV ~10, the MC-AS is densely populated by axons ( C ). Applying Alexa Fluor 594 in the MC-AS demonstrate fluidic isolation. Inset shows calcein fluorescent axons. ( D, E ). In the absence of cells (no cells), T3 I125 is applied in the MC-AS, and only background radioactivity was detected in the MC-CS. Chromatograms from the MC-AS (orange) show typical peaks of T3 I125 T2 I125 , and I I125 .

Article Snippet: We used a microfluidic compartmentalized culture device that contains 450 μM long microchannels connecting MC-CS and MC-AS and permits only distal axons to grow into the MC-AS (XonaChip, Cat# XC450, Xona Microfluidics Temecula, CA, USA).

Techniques: Isolation, Radioactivity