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A) Immunofluorescence staining of primary neurons derived from mouse cortex. NSE (green) and GFAP (red) coexistence indicates the presence of both glial cells and primary neurons (n=3 independent experiments). B) Primary neurons co-stained with MAP2 and Phalloidin to observe postsynaptic spines in neuronal dendrites. Scale bar 10 microns (n=3 independent experiments). C) MAP2 and Phalloidin staining shows the presence of mushroom (a), stubby (b) and filopodia (c) spines in B. D) Co-staining of the presynaptic marker synaptophysin (Syp) and the postsynaptic marker PSD95 indicates the presence of active glutamatergic synapses. (n=3 independent experiments). E) Overlapping profile of the co-staining synaptophysin and PSD95 shown in D. A.U. is arbitrary units. F) Timelapse microscopy showing the overexpression of the volume marker mCherry and the pH-sensitive <t>CD63-pHluorin</t> plasmid as an indicator of MVB fusion to the plasma membrane and indirectly suggesting the release of exosomes from cell bodies and dendrites of cortical neurons. Scale bar 10 microns. (n=3 independent experiments). G) Magnification of region indicated in F. The star indicates a permanent patch of CD63-pHluorin in the plasma membrane, the arrow a brief exosomal release event.
Cd63 Phluorin Construct, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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A) Immunofluorescence staining of primary neurons derived from mouse cortex. NSE (green) and GFAP (red) coexistence indicates the presence of both glial cells and primary neurons (n=3 independent experiments). B) Primary neurons co-stained with MAP2 and Phalloidin to observe postsynaptic spines in neuronal dendrites. Scale bar 10 microns (n=3 independent experiments). C) MAP2 and Phalloidin staining shows the presence of mushroom (a), stubby (b) and filopodia (c) spines in B. D) Co-staining of the presynaptic marker synaptophysin (Syp) and the postsynaptic marker PSD95 indicates the presence of active glutamatergic synapses. (n=3 independent experiments). E) Overlapping profile of the co-staining synaptophysin and PSD95 shown in D. A.U. is arbitrary units. F) Timelapse microscopy showing the overexpression of the volume marker mCherry and the pH-sensitive <t>CD63-pHluorin</t> plasmid as an indicator of MVB fusion to the plasma membrane and indirectly suggesting the release of exosomes from cell bodies and dendrites of cortical neurons. Scale bar 10 microns. (n=3 independent experiments). G) Magnification of region indicated in F. The star indicates a permanent patch of CD63-pHluorin in the plasma membrane, the arrow a brief exosomal release event.
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A) Immunofluorescence staining of primary neurons derived from mouse cortex. NSE (green) and GFAP (red) coexistence indicates the presence of both glial cells and primary neurons (n=3 independent experiments). B) Primary neurons co-stained with MAP2 and Phalloidin to observe postsynaptic spines in neuronal dendrites. Scale bar 10 microns (n=3 independent experiments). C) MAP2 and Phalloidin staining shows the presence of mushroom (a), stubby (b) and filopodia (c) spines in B. D) Co-staining of the presynaptic marker synaptophysin (Syp) and the postsynaptic marker PSD95 indicates the presence of active glutamatergic synapses. (n=3 independent experiments). E) Overlapping profile of the co-staining synaptophysin and PSD95 shown in D. A.U. is arbitrary units. F) Timelapse microscopy showing the overexpression of the volume marker mCherry and the pH-sensitive <t>CD63-pHluorin</t> plasmid as an indicator of MVB fusion to the plasma membrane and indirectly suggesting the release of exosomes from cell bodies and dendrites of cortical neurons. Scale bar 10 microns. (n=3 independent experiments). G) Magnification of region indicated in F. The star indicates a permanent patch of CD63-pHluorin in the plasma membrane, the arrow a brief exosomal release event.
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A) Immunofluorescence staining of primary neurons derived from mouse cortex. NSE (green) and GFAP (red) coexistence indicates the presence of both glial cells and primary neurons (n=3 independent experiments). B) Primary neurons co-stained with MAP2 and Phalloidin to observe postsynaptic spines in neuronal dendrites. Scale bar 10 microns (n=3 independent experiments). C) MAP2 and Phalloidin staining shows the presence of mushroom (a), stubby (b) and filopodia (c) spines in B. D) Co-staining of the presynaptic marker synaptophysin (Syp) and the postsynaptic marker PSD95 indicates the presence of active glutamatergic synapses. (n=3 independent experiments). E) Overlapping profile of the co-staining synaptophysin and PSD95 shown in D. A.U. is arbitrary units. F) Timelapse microscopy showing the overexpression of the volume marker mCherry and the pH-sensitive <t>CD63-pHluorin</t> plasmid as an indicator of MVB fusion to the plasma membrane and indirectly suggesting the release of exosomes from cell bodies and dendrites of cortical neurons. Scale bar 10 microns. (n=3 independent experiments). G) Magnification of region indicated in F. The star indicates a permanent patch of CD63-pHluorin in the plasma membrane, the arrow a brief exosomal release event.
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A) Immunofluorescence staining of primary neurons derived from mouse cortex. NSE (green) and GFAP (red) coexistence indicates the presence of both glial cells and primary neurons (n=3 independent experiments). B) Primary neurons co-stained with MAP2 and Phalloidin to observe postsynaptic spines in neuronal dendrites. Scale bar 10 microns (n=3 independent experiments). C) MAP2 and Phalloidin staining shows the presence of mushroom (a), stubby (b) and filopodia (c) spines in B. D) Co-staining of the presynaptic marker synaptophysin (Syp) and the postsynaptic marker PSD95 indicates the presence of active glutamatergic synapses. (n=3 independent experiments). E) Overlapping profile of the co-staining synaptophysin and PSD95 shown in D. A.U. is arbitrary units. F) Timelapse microscopy showing the overexpression of the volume marker mCherry and the pH-sensitive <t>CD63-pHluorin</t> plasmid as an indicator of MVB fusion to the plasma membrane and indirectly suggesting the release of exosomes from cell bodies and dendrites of cortical neurons. Scale bar 10 microns. (n=3 independent experiments). G) Magnification of region indicated in F. The star indicates a permanent patch of CD63-pHluorin in the plasma membrane, the arrow a brief exosomal release event.
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A) Immunofluorescence staining of primary neurons derived from mouse cortex. NSE (green) and GFAP (red) coexistence indicates the presence of both glial cells and primary neurons (n=3 independent experiments). B) Primary neurons co-stained with MAP2 and Phalloidin to observe postsynaptic spines in neuronal dendrites. Scale bar 10 microns (n=3 independent experiments). C) MAP2 and Phalloidin staining shows the presence of mushroom (a), stubby (b) and filopodia (c) spines in B. D) Co-staining of the presynaptic marker synaptophysin (Syp) and the postsynaptic marker PSD95 indicates the presence of active glutamatergic synapses. (n=3 independent experiments). E) Overlapping profile of the co-staining synaptophysin and PSD95 shown in D. A.U. is arbitrary units. F) Timelapse microscopy showing the overexpression of the volume marker mCherry and the pH-sensitive <t>CD63-pHluorin</t> plasmid as an indicator of MVB fusion to the plasma membrane and indirectly suggesting the release of exosomes from cell bodies and dendrites of cortical neurons. Scale bar 10 microns. (n=3 independent experiments). G) Magnification of region indicated in F. The star indicates a permanent patch of CD63-pHluorin in the plasma membrane, the arrow a brief exosomal release event.
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A) Immunofluorescence staining of primary neurons derived from mouse cortex. NSE (green) and GFAP (red) coexistence indicates the presence of both glial cells and primary neurons (n=3 independent experiments). B) Primary neurons co-stained with MAP2 and Phalloidin to observe postsynaptic spines in neuronal dendrites. Scale bar 10 microns (n=3 independent experiments). C) MAP2 and Phalloidin staining shows the presence of mushroom (a), stubby (b) and filopodia (c) spines in B. D) Co-staining of the presynaptic marker synaptophysin (Syp) and the postsynaptic marker PSD95 indicates the presence of active glutamatergic synapses. (n=3 independent experiments). E) Overlapping profile of the co-staining synaptophysin and PSD95 shown in D. A.U. is arbitrary units. F) Timelapse microscopy showing the overexpression of the volume marker mCherry and the pH-sensitive CD63-pHluorin plasmid as an indicator of MVB fusion to the plasma membrane and indirectly suggesting the release of exosomes from cell bodies and dendrites of cortical neurons. Scale bar 10 microns. (n=3 independent experiments). G) Magnification of region indicated in F. The star indicates a permanent patch of CD63-pHluorin in the plasma membrane, the arrow a brief exosomal release event.

Journal: bioRxiv

Article Title: Extracellular vesicles released from cortical neurons influence spontaneous activity of recipient neurons

doi: 10.1101/2025.04.09.647922

Figure Lengend Snippet: A) Immunofluorescence staining of primary neurons derived from mouse cortex. NSE (green) and GFAP (red) coexistence indicates the presence of both glial cells and primary neurons (n=3 independent experiments). B) Primary neurons co-stained with MAP2 and Phalloidin to observe postsynaptic spines in neuronal dendrites. Scale bar 10 microns (n=3 independent experiments). C) MAP2 and Phalloidin staining shows the presence of mushroom (a), stubby (b) and filopodia (c) spines in B. D) Co-staining of the presynaptic marker synaptophysin (Syp) and the postsynaptic marker PSD95 indicates the presence of active glutamatergic synapses. (n=3 independent experiments). E) Overlapping profile of the co-staining synaptophysin and PSD95 shown in D. A.U. is arbitrary units. F) Timelapse microscopy showing the overexpression of the volume marker mCherry and the pH-sensitive CD63-pHluorin plasmid as an indicator of MVB fusion to the plasma membrane and indirectly suggesting the release of exosomes from cell bodies and dendrites of cortical neurons. Scale bar 10 microns. (n=3 independent experiments). G) Magnification of region indicated in F. The star indicates a permanent patch of CD63-pHluorin in the plasma membrane, the arrow a brief exosomal release event.

Article Snippet: Cells were transfected at DIV 6-7 with Calcium Phosphate method using CD63-pHluorin construct from addgene (#130901) and mCherry as a volume marker.

Techniques: Immunofluorescence, Staining, Derivative Assay, Marker, Microscopy, Over Expression, Plasmid Preparation, Membrane

A) Glutamate receptor subunits are present in EVs isolated from primary cultures, in combination with the EV-marker CD63. B) Immunoelectron microscopy of EVs isolated from cultured neurons support the presence of the glutamate receptor subunit GluA2. C) EVs isolated from mouse brains and validated qualitatively by the presence of the EV markers Flotillin-1 and CD81, contain both AMPA receptor subunits and Nr2b subunit of NMDA receptors.

Journal: bioRxiv

Article Title: Extracellular vesicles released from cortical neurons influence spontaneous activity of recipient neurons

doi: 10.1101/2025.04.09.647922

Figure Lengend Snippet: A) Glutamate receptor subunits are present in EVs isolated from primary cultures, in combination with the EV-marker CD63. B) Immunoelectron microscopy of EVs isolated from cultured neurons support the presence of the glutamate receptor subunit GluA2. C) EVs isolated from mouse brains and validated qualitatively by the presence of the EV markers Flotillin-1 and CD81, contain both AMPA receptor subunits and Nr2b subunit of NMDA receptors.

Article Snippet: Cells were transfected at DIV 6-7 with Calcium Phosphate method using CD63-pHluorin construct from addgene (#130901) and mCherry as a volume marker.

Techniques: Isolation, Marker, Immuno-Electron Microscopy, Cell Culture