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(a) Scheme of the experimental design for tau aggregate treatment in mixed cultures. (b) Micrographs of a large section of culture depicting neurospheroids in bighfield. Examples of stainings for nucleis, glial and neuronal markers (Dapi, GFAP, NeuN, NFL). (c-e) Bar plots of the quantification of neuropheroid density (c), as well as NeuN (d) and neurofilament (e) coverage of neurospheroids. (f) Illustration of neurospheroid stained for synaptic markers with example of an analyzed volume depicted with a white rectangle and illustrated in the center. Overlayed white objects (e.g. white arrow) correspond to the identified active zones. On the right, a small diagram illustrating how the active zone object were reconstructed from PSD-95 and vGlut1 signals. (g) Detailed examples of pre and post synaptic markers images (top) and associated 3D reconstruction (bottom). (h-j) Bar plots of the total PSD-95 (h) and vGlut1 (i) volumes (medians), and active zone volume per surface of <t>PSD95.</t> (N_=_3 cultures, total of 25 neurospheroids scanned, and 5 neurites VOIs analyzed per image; *** p <0.001).
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(a) Scheme of the experimental design for tau aggregate treatment in mixed cultures. (b) Micrographs of a large section of culture depicting neurospheroids in bighfield. Examples of stainings for nucleis, glial and neuronal markers (Dapi, GFAP, NeuN, NFL). (c-e) Bar plots of the quantification of neuropheroid density (c), as well as NeuN (d) and neurofilament (e) coverage of neurospheroids. (f) Illustration of neurospheroid stained for synaptic markers with example of an analyzed volume depicted with a white rectangle and illustrated in the center. Overlayed white objects (e.g. white arrow) correspond to the identified active zones. On the right, a small diagram illustrating how the active zone object were reconstructed from PSD-95 and vGlut1 signals. (g) Detailed examples of pre and post synaptic markers images (top) and associated 3D reconstruction (bottom). (h-j) Bar plots of the total PSD-95 (h) and vGlut1 (i) volumes (medians), and active zone volume per surface of <t>PSD95.</t> (N_=_3 cultures, total of 25 neurospheroids scanned, and 5 neurites VOIs analyzed per image; *** p <0.001).
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(a) Scheme of the experimental design for tau aggregate treatment in mixed cultures. (b) Micrographs of a large section of culture depicting neurospheroids in bighfield. Examples of stainings for nucleis, glial and neuronal markers (Dapi, GFAP, NeuN, NFL). (c-e) Bar plots of the quantification of neuropheroid density (c), as well as NeuN (d) and neurofilament (e) coverage of neurospheroids. (f) Illustration of neurospheroid stained for synaptic markers with example of an analyzed volume depicted with a white rectangle and illustrated in the center. Overlayed white objects (e.g. white arrow) correspond to the identified active zones. On the right, a small diagram illustrating how the active zone object were reconstructed from PSD-95 and vGlut1 signals. (g) Detailed examples of pre and post synaptic markers images (top) and associated 3D reconstruction (bottom). (h-j) Bar plots of the total PSD-95 (h) and vGlut1 (i) volumes (medians), and active zone volume per surface of <t>PSD95.</t> (N_=_3 cultures, total of 25 neurospheroids scanned, and 5 neurites VOIs analyzed per image; *** p <0.001).
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(a) Scheme of the experimental design for tau aggregate treatment in mixed cultures. (b) Micrographs of a large section of culture depicting neurospheroids in bighfield. Examples of stainings for nucleis, glial and neuronal markers (Dapi, GFAP, NeuN, NFL). (c-e) Bar plots of the quantification of neuropheroid density (c), as well as NeuN (d) and neurofilament (e) coverage of neurospheroids. (f) Illustration of neurospheroid stained for synaptic markers with example of an analyzed volume depicted with a white rectangle and illustrated in the center. Overlayed white objects (e.g. white arrow) correspond to the identified active zones. On the right, a small diagram illustrating how the active zone object were reconstructed from PSD-95 and vGlut1 signals. (g) Detailed examples of pre and post synaptic markers images (top) and associated 3D reconstruction (bottom). (h-j) Bar plots of the total PSD-95 (h) and vGlut1 (i) volumes (medians), and active zone volume per surface of <t>PSD95.</t> (N_=_3 cultures, total of 25 neurospheroids scanned, and 5 neurites VOIs analyzed per image; *** p <0.001).
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(a) Scheme of the experimental design for tau aggregate treatment in mixed cultures. (b) Micrographs of a large section of culture depicting neurospheroids in bighfield. Examples of stainings for nucleis, glial and neuronal markers (Dapi, GFAP, NeuN, NFL). (c-e) Bar plots of the quantification of neuropheroid density (c), as well as NeuN (d) and neurofilament (e) coverage of neurospheroids. (f) Illustration of neurospheroid stained for synaptic markers with example of an analyzed volume depicted with a white rectangle and illustrated in the center. Overlayed white objects (e.g. white arrow) correspond to the identified active zones. On the right, a small diagram illustrating how the active zone object were reconstructed from PSD-95 and vGlut1 signals. (g) Detailed examples of pre and post synaptic markers images (top) and associated 3D reconstruction (bottom). (h-j) Bar plots of the total PSD-95 (h) and vGlut1 (i) volumes (medians), and active zone volume per surface of PSD95. (N_=_3 cultures, total of 25 neurospheroids scanned, and 5 neurites VOIs analyzed per image; *** p <0.001).

Journal: bioRxiv

Article Title: Inverse and Postponed Impacts of Extracellular Tau PHF on Astrocytes and Neurons’ Mitochondrial Function

doi: 10.1101/2024.03.19.585791

Figure Lengend Snippet: (a) Scheme of the experimental design for tau aggregate treatment in mixed cultures. (b) Micrographs of a large section of culture depicting neurospheroids in bighfield. Examples of stainings for nucleis, glial and neuronal markers (Dapi, GFAP, NeuN, NFL). (c-e) Bar plots of the quantification of neuropheroid density (c), as well as NeuN (d) and neurofilament (e) coverage of neurospheroids. (f) Illustration of neurospheroid stained for synaptic markers with example of an analyzed volume depicted with a white rectangle and illustrated in the center. Overlayed white objects (e.g. white arrow) correspond to the identified active zones. On the right, a small diagram illustrating how the active zone object were reconstructed from PSD-95 and vGlut1 signals. (g) Detailed examples of pre and post synaptic markers images (top) and associated 3D reconstruction (bottom). (h-j) Bar plots of the total PSD-95 (h) and vGlut1 (i) volumes (medians), and active zone volume per surface of PSD95. (N_=_3 cultures, total of 25 neurospheroids scanned, and 5 neurites VOIs analyzed per image; *** p <0.001).

Article Snippet: Primary antibody incubation was performed overnight with rabbit anti-VGlut1 (Cell Signaling Technology, 1230.31; 1:500), chicken anti MAP2 (Abcam, ab5392, 1:1’000), goat anti-PSD95 (Abcam, ab12093; 1:500), or mouse anti-NeuN (Chemicon, MAB337) and anti-GFAP (Dako, GA52461-2) antibodies.

Techniques: Staining