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psd95 puncta  (Oxford Instruments)


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    Structured Review

    Oxford Instruments psd95 puncta
    ( A to C ) Representative images (A), quantification of protrusion density (B), and mushroom spine, stubby spine, thin spine, and filopodia density (C) of DIV-18 primary hippocampal neurons transfected with scramble control or circHomer1 shRNA ( n = 10 neurons per group from three independent experiments; protrusion density, mushroom, stubby, and thin: Student’s t test, filopodia: Mann-Whitney test). Arrow colors indicate mushroom (white), stubby (yellow), thin (green), and filopodia (blue), respectively. Scale bars, 50 μm (up) and 5 μm (down). ( D and E ) Representative images (D) and quantification (E) of <t>PSD95.FingR</t> expression in DIV-18 primary hippocampal neurons transfected with scramble control or circHomer1 shRNA ( n = 10 neurons per group from three independent experiments; Student’s t test). White arrows indicate PSD95.FingR signals at dendritic spines. Scale bar, 5 μm. ( F to H ) Representative images (F), quantification of protrusion density (G), and mushroom spine, stubby spine, thin spine, and filopodia density (H) of DIV-18 primary hippocampal neurons transfected with vector control or circHomer1 overexpression vector ( n = 11 neurons per group from three independent experiments; protrusion density, mushroom, stubby, and thin: Student’s t test, filopodia: Mann-Whitney test). Arrow colors indicate mushroom (white), stubby (yellow), thin (green), and filopodia (blue), respectively. Scale bars, 50 μm (up) and 5 μm (down). O.E., overexpression. ( I and J ) Representative images (I) and quantification (J) of PSD95.FingR expression in DIV-18 primary hippocampal neurons transfected with vector control or circHomer1 overexpression vector ( n = 9 to 10 per group from three independent experiments; Student’s t test). White arrows indicate PSD95.FingR signals at dendritic spines. Scale bar, 5 μm. Data are mean ± SEM. * P < 0.05, ** P < 0.01, and *** P < 0.001.
    Psd95 Puncta, supplied by Oxford Instruments, used in various techniques. Bioz Stars score: 99/100, based on 44266 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/psd95+puncta/Imaris/pmc12893322-324-6-11
    Average 99 stars, based on 44266 article reviews
    psd95 puncta - by Bioz Stars, 2026-09
    99/100 stars

    Images

    1) Product Images from "Circular RNA circHomer1 mediates hippocampal functions via ribonucleoprotein granule transport and dendritic targeting of synaptic RNAs"

    Article Title: Circular RNA circHomer1 mediates hippocampal functions via ribonucleoprotein granule transport and dendritic targeting of synaptic RNAs

    Journal: Science Advances

    doi: 10.1126/sciadv.ads7509

    ( A to C ) Representative images (A), quantification of protrusion density (B), and mushroom spine, stubby spine, thin spine, and filopodia density (C) of DIV-18 primary hippocampal neurons transfected with scramble control or circHomer1 shRNA ( n = 10 neurons per group from three independent experiments; protrusion density, mushroom, stubby, and thin: Student’s t test, filopodia: Mann-Whitney test). Arrow colors indicate mushroom (white), stubby (yellow), thin (green), and filopodia (blue), respectively. Scale bars, 50 μm (up) and 5 μm (down). ( D and E ) Representative images (D) and quantification (E) of PSD95.FingR expression in DIV-18 primary hippocampal neurons transfected with scramble control or circHomer1 shRNA ( n = 10 neurons per group from three independent experiments; Student’s t test). White arrows indicate PSD95.FingR signals at dendritic spines. Scale bar, 5 μm. ( F to H ) Representative images (F), quantification of protrusion density (G), and mushroom spine, stubby spine, thin spine, and filopodia density (H) of DIV-18 primary hippocampal neurons transfected with vector control or circHomer1 overexpression vector ( n = 11 neurons per group from three independent experiments; protrusion density, mushroom, stubby, and thin: Student’s t test, filopodia: Mann-Whitney test). Arrow colors indicate mushroom (white), stubby (yellow), thin (green), and filopodia (blue), respectively. Scale bars, 50 μm (up) and 5 μm (down). O.E., overexpression. ( I and J ) Representative images (I) and quantification (J) of PSD95.FingR expression in DIV-18 primary hippocampal neurons transfected with vector control or circHomer1 overexpression vector ( n = 9 to 10 per group from three independent experiments; Student’s t test). White arrows indicate PSD95.FingR signals at dendritic spines. Scale bar, 5 μm. Data are mean ± SEM. * P < 0.05, ** P < 0.01, and *** P < 0.001.
    Figure Legend Snippet: ( A to C ) Representative images (A), quantification of protrusion density (B), and mushroom spine, stubby spine, thin spine, and filopodia density (C) of DIV-18 primary hippocampal neurons transfected with scramble control or circHomer1 shRNA ( n = 10 neurons per group from three independent experiments; protrusion density, mushroom, stubby, and thin: Student’s t test, filopodia: Mann-Whitney test). Arrow colors indicate mushroom (white), stubby (yellow), thin (green), and filopodia (blue), respectively. Scale bars, 50 μm (up) and 5 μm (down). ( D and E ) Representative images (D) and quantification (E) of PSD95.FingR expression in DIV-18 primary hippocampal neurons transfected with scramble control or circHomer1 shRNA ( n = 10 neurons per group from three independent experiments; Student’s t test). White arrows indicate PSD95.FingR signals at dendritic spines. Scale bar, 5 μm. ( F to H ) Representative images (F), quantification of protrusion density (G), and mushroom spine, stubby spine, thin spine, and filopodia density (H) of DIV-18 primary hippocampal neurons transfected with vector control or circHomer1 overexpression vector ( n = 11 neurons per group from three independent experiments; protrusion density, mushroom, stubby, and thin: Student’s t test, filopodia: Mann-Whitney test). Arrow colors indicate mushroom (white), stubby (yellow), thin (green), and filopodia (blue), respectively. Scale bars, 50 μm (up) and 5 μm (down). O.E., overexpression. ( I and J ) Representative images (I) and quantification (J) of PSD95.FingR expression in DIV-18 primary hippocampal neurons transfected with vector control or circHomer1 overexpression vector ( n = 9 to 10 per group from three independent experiments; Student’s t test). White arrows indicate PSD95.FingR signals at dendritic spines. Scale bar, 5 μm. Data are mean ± SEM. * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Techniques Used: Transfection, Control, shRNA, MANN-WHITNEY, Expressing, Plasmid Preparation, Over Expression

    ( A ) eMAP workflow. ( B ) Representative images showing the dendrites of pyramidal neurons in the CA1 region of the 11-week-old mouse hippocampus. Arrow colors indicate different types of dendritic spines. Scale bars, 100 μm (left) and 10 μm (right), estimated to be 33.33- and 3.33-μm preexpansion, respectively. ( C and D ) Quantification of protrusion density (C), mushroom spine, stubby spine, thin spine, and filopodia density (D) at dendrites ( n = 22 to 29 dendrites from five mice per group; protrusion density, mushroom, and thin: Student’s t test, stubby and filopodia: Mann-Whitney test). ( E ) Representative images showing 3D reconstructions of dendrites. Scale bar, 10 μm, estimated to be 3.33-μm preexpansion. ( F and G ) Quantification (F) and distribution (G) of dendritic spine volume ( n = 699 to 716 dendritic spines from five mice per group; 25th, 50th, and 75th percentiles of the data are presented; Mann-Whitney test). ( H ) Representative images showing fluorescence images and 3D reconstructions of PSD95 puncta (red) at dendritic spines. Scale bar, 5 μm, estimated to be 1.67-μm preexpansion. ( I and J ) Quantification (I) and distribution (J) of PSD95 puncta volume at dendritic spines ( n = 280 to 304 PSD95 puncta from five mice per group; 25th, 50th, and 75th percentiles of the data are presented; Mann-Whitney test). ( K ) A representative image illustrating whole-cell patch clamp recordings from CA1 stratum radiatum neurons in acute hippocampal brain slices. ( L ) Representative traces of spontaneous excitatory postsynaptic currents (sEPSCs). ( M ) Quantification of the frequency sEPSCs ( n = 9 to 11 cells per group; average sEPSC frequency: Student’s t test, cumulative fraction: Kolmogorov-Smirnov test). Values in (C) and (D), (F) and (G), and (I) and (J) were normalized to 3× expansion factor. Data are mean ± SEM unless otherwise specified. * P < 0.05 and *** P < 0.001.
    Figure Legend Snippet: ( A ) eMAP workflow. ( B ) Representative images showing the dendrites of pyramidal neurons in the CA1 region of the 11-week-old mouse hippocampus. Arrow colors indicate different types of dendritic spines. Scale bars, 100 μm (left) and 10 μm (right), estimated to be 33.33- and 3.33-μm preexpansion, respectively. ( C and D ) Quantification of protrusion density (C), mushroom spine, stubby spine, thin spine, and filopodia density (D) at dendrites ( n = 22 to 29 dendrites from five mice per group; protrusion density, mushroom, and thin: Student’s t test, stubby and filopodia: Mann-Whitney test). ( E ) Representative images showing 3D reconstructions of dendrites. Scale bar, 10 μm, estimated to be 3.33-μm preexpansion. ( F and G ) Quantification (F) and distribution (G) of dendritic spine volume ( n = 699 to 716 dendritic spines from five mice per group; 25th, 50th, and 75th percentiles of the data are presented; Mann-Whitney test). ( H ) Representative images showing fluorescence images and 3D reconstructions of PSD95 puncta (red) at dendritic spines. Scale bar, 5 μm, estimated to be 1.67-μm preexpansion. ( I and J ) Quantification (I) and distribution (J) of PSD95 puncta volume at dendritic spines ( n = 280 to 304 PSD95 puncta from five mice per group; 25th, 50th, and 75th percentiles of the data are presented; Mann-Whitney test). ( K ) A representative image illustrating whole-cell patch clamp recordings from CA1 stratum radiatum neurons in acute hippocampal brain slices. ( L ) Representative traces of spontaneous excitatory postsynaptic currents (sEPSCs). ( M ) Quantification of the frequency sEPSCs ( n = 9 to 11 cells per group; average sEPSC frequency: Student’s t test, cumulative fraction: Kolmogorov-Smirnov test). Values in (C) and (D), (F) and (G), and (I) and (J) were normalized to 3× expansion factor. Data are mean ± SEM unless otherwise specified. * P < 0.05 and *** P < 0.001.

    Techniques Used: MANN-WHITNEY, Fluorescence, Patch Clamp

    Related Articles

    Transfection:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    Control:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    shRNA:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    MANN-WHITNEY:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    Expressing:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    Plasmid Preparation:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    Over Expression:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    Fluorescence:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    Patch Clamp:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    Labeling:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    Isolation:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    Comparison:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    Immunohistochemistry:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    Staining:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    Saline:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    Virus:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.

    Eye Drops:

    Article Title: Triggering Receptor Expressed on Myeloid Cells 2 Alleviated Sevoflurane-Induced Developmental Neurotoxicity via Microglial Pruning of Dendritic Spines in the CA1 Region of the Hippocampus.
    Article Snippet: Sevoflurane induces developmental neurotoxicity in mice; however, the underlying mechanisms remain unclear.. Triggering receptor expressed on myeloid cells 2 (TREM2) is essential for microglia-mediated synaptic refinement during the early stages of brain development.. We explored the effects of TREM2 on dendritic spine pruning during sevoflurane-induced developmental neurotoxicity in mice.



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    ( A to C ) Representative images (A), quantification of protrusion density (B), and mushroom spine, stubby spine, thin spine, and filopodia density (C) of DIV-18 primary hippocampal neurons transfected with scramble control or circHomer1 shRNA ( n = 10 neurons per group from three independent experiments; protrusion density, mushroom, stubby, and thin: Student’s t test, filopodia: Mann-Whitney test). Arrow colors indicate mushroom (white), stubby (yellow), thin (green), and filopodia (blue), respectively. Scale bars, 50 μm (up) and 5 μm (down). ( D and E ) Representative images (D) and quantification (E) of <t>PSD95.FingR</t> expression in DIV-18 primary hippocampal neurons transfected with scramble control or circHomer1 shRNA ( n = 10 neurons per group from three independent experiments; Student’s t test). White arrows indicate PSD95.FingR signals at dendritic spines. Scale bar, 5 μm. ( F to H ) Representative images (F), quantification of protrusion density (G), and mushroom spine, stubby spine, thin spine, and filopodia density (H) of DIV-18 primary hippocampal neurons transfected with vector control or circHomer1 overexpression vector ( n = 11 neurons per group from three independent experiments; protrusion density, mushroom, stubby, and thin: Student’s t test, filopodia: Mann-Whitney test). Arrow colors indicate mushroom (white), stubby (yellow), thin (green), and filopodia (blue), respectively. Scale bars, 50 μm (up) and 5 μm (down). O.E., overexpression. ( I and J ) Representative images (I) and quantification (J) of PSD95.FingR expression in DIV-18 primary hippocampal neurons transfected with vector control or circHomer1 overexpression vector ( n = 9 to 10 per group from three independent experiments; Student’s t test). White arrows indicate PSD95.FingR signals at dendritic spines. Scale bar, 5 μm. Data are mean ± SEM. * P < 0.05, ** P < 0.01, and *** P < 0.001.
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    ( A ) <t>DiO-PSD95.FingR-Citrine</t> was injected in barrel cortex of SST-Cre mice. ( B ) Schematic outlining injection and training timeline. Animals in fixed tissue analysis were time matched to control for viral expression time. ( C ) 10x image of SST-Cre mice expressing PSD95.FingR across the cortical column. Scale=100µm ( D ) 63x volumetric image of PSD95.FingR labeled somatostatin neurons. Scale=10µm ( E ) (Left) Zoomed image of PSD95.FingR puncta along a dendrite. (Right) Surface reconstruction of citrine signal. Scale=1µm ( F ) Immunohistochemical labeling of SST neurons (Left) PSD95.FingR-Citrine (Middle) calretinin expression (protein product of Calb2 gene) Scale=10µm (Right) merged channels. ( G ) Schematic indicating layer imaging field of view was captured (Layer 2/3). ( H ) Cumulative distribution of PSD95 puncta volume in layer 2/3 SST-O neurons. ( I ) Same as (H) for L2/3 SST-Calb2 neurons. ( J ) Within animal comparison of mean PSD95 puncta volume between SST-O (gray bars) and SST-Calb2 (pink bars) after experiencing acclimation (ACC), one, or five days of sensory association training (SAT). ( K-N ) Same as G-J but for L5 SST-O and SST-Calb2 SST neurons.
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    Image Search Results


    ( A to C ) Representative images (A), quantification of protrusion density (B), and mushroom spine, stubby spine, thin spine, and filopodia density (C) of DIV-18 primary hippocampal neurons transfected with scramble control or circHomer1 shRNA ( n = 10 neurons per group from three independent experiments; protrusion density, mushroom, stubby, and thin: Student’s t test, filopodia: Mann-Whitney test). Arrow colors indicate mushroom (white), stubby (yellow), thin (green), and filopodia (blue), respectively. Scale bars, 50 μm (up) and 5 μm (down). ( D and E ) Representative images (D) and quantification (E) of PSD95.FingR expression in DIV-18 primary hippocampal neurons transfected with scramble control or circHomer1 shRNA ( n = 10 neurons per group from three independent experiments; Student’s t test). White arrows indicate PSD95.FingR signals at dendritic spines. Scale bar, 5 μm. ( F to H ) Representative images (F), quantification of protrusion density (G), and mushroom spine, stubby spine, thin spine, and filopodia density (H) of DIV-18 primary hippocampal neurons transfected with vector control or circHomer1 overexpression vector ( n = 11 neurons per group from three independent experiments; protrusion density, mushroom, stubby, and thin: Student’s t test, filopodia: Mann-Whitney test). Arrow colors indicate mushroom (white), stubby (yellow), thin (green), and filopodia (blue), respectively. Scale bars, 50 μm (up) and 5 μm (down). O.E., overexpression. ( I and J ) Representative images (I) and quantification (J) of PSD95.FingR expression in DIV-18 primary hippocampal neurons transfected with vector control or circHomer1 overexpression vector ( n = 9 to 10 per group from three independent experiments; Student’s t test). White arrows indicate PSD95.FingR signals at dendritic spines. Scale bar, 5 μm. Data are mean ± SEM. * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Journal: Science Advances

    Article Title: Circular RNA circHomer1 mediates hippocampal functions via ribonucleoprotein granule transport and dendritic targeting of synaptic RNAs

    doi: 10.1126/sciadv.ads7509

    Figure Lengend Snippet: ( A to C ) Representative images (A), quantification of protrusion density (B), and mushroom spine, stubby spine, thin spine, and filopodia density (C) of DIV-18 primary hippocampal neurons transfected with scramble control or circHomer1 shRNA ( n = 10 neurons per group from three independent experiments; protrusion density, mushroom, stubby, and thin: Student’s t test, filopodia: Mann-Whitney test). Arrow colors indicate mushroom (white), stubby (yellow), thin (green), and filopodia (blue), respectively. Scale bars, 50 μm (up) and 5 μm (down). ( D and E ) Representative images (D) and quantification (E) of PSD95.FingR expression in DIV-18 primary hippocampal neurons transfected with scramble control or circHomer1 shRNA ( n = 10 neurons per group from three independent experiments; Student’s t test). White arrows indicate PSD95.FingR signals at dendritic spines. Scale bar, 5 μm. ( F to H ) Representative images (F), quantification of protrusion density (G), and mushroom spine, stubby spine, thin spine, and filopodia density (H) of DIV-18 primary hippocampal neurons transfected with vector control or circHomer1 overexpression vector ( n = 11 neurons per group from three independent experiments; protrusion density, mushroom, stubby, and thin: Student’s t test, filopodia: Mann-Whitney test). Arrow colors indicate mushroom (white), stubby (yellow), thin (green), and filopodia (blue), respectively. Scale bars, 50 μm (up) and 5 μm (down). O.E., overexpression. ( I and J ) Representative images (I) and quantification (J) of PSD95.FingR expression in DIV-18 primary hippocampal neurons transfected with vector control or circHomer1 overexpression vector ( n = 9 to 10 per group from three independent experiments; Student’s t test). White arrows indicate PSD95.FingR signals at dendritic spines. Scale bar, 5 μm. Data are mean ± SEM. * P < 0.05, ** P < 0.01, and *** P < 0.001.

    Article Snippet: Volumetric analysis of dendritic spines and PSD95 puncta was performed using Imaris (version 10.0).

    Techniques: Transfection, Control, shRNA, MANN-WHITNEY, Expressing, Plasmid Preparation, Over Expression

    ( A ) eMAP workflow. ( B ) Representative images showing the dendrites of pyramidal neurons in the CA1 region of the 11-week-old mouse hippocampus. Arrow colors indicate different types of dendritic spines. Scale bars, 100 μm (left) and 10 μm (right), estimated to be 33.33- and 3.33-μm preexpansion, respectively. ( C and D ) Quantification of protrusion density (C), mushroom spine, stubby spine, thin spine, and filopodia density (D) at dendrites ( n = 22 to 29 dendrites from five mice per group; protrusion density, mushroom, and thin: Student’s t test, stubby and filopodia: Mann-Whitney test). ( E ) Representative images showing 3D reconstructions of dendrites. Scale bar, 10 μm, estimated to be 3.33-μm preexpansion. ( F and G ) Quantification (F) and distribution (G) of dendritic spine volume ( n = 699 to 716 dendritic spines from five mice per group; 25th, 50th, and 75th percentiles of the data are presented; Mann-Whitney test). ( H ) Representative images showing fluorescence images and 3D reconstructions of PSD95 puncta (red) at dendritic spines. Scale bar, 5 μm, estimated to be 1.67-μm preexpansion. ( I and J ) Quantification (I) and distribution (J) of PSD95 puncta volume at dendritic spines ( n = 280 to 304 PSD95 puncta from five mice per group; 25th, 50th, and 75th percentiles of the data are presented; Mann-Whitney test). ( K ) A representative image illustrating whole-cell patch clamp recordings from CA1 stratum radiatum neurons in acute hippocampal brain slices. ( L ) Representative traces of spontaneous excitatory postsynaptic currents (sEPSCs). ( M ) Quantification of the frequency sEPSCs ( n = 9 to 11 cells per group; average sEPSC frequency: Student’s t test, cumulative fraction: Kolmogorov-Smirnov test). Values in (C) and (D), (F) and (G), and (I) and (J) were normalized to 3× expansion factor. Data are mean ± SEM unless otherwise specified. * P < 0.05 and *** P < 0.001.

    Journal: Science Advances

    Article Title: Circular RNA circHomer1 mediates hippocampal functions via ribonucleoprotein granule transport and dendritic targeting of synaptic RNAs

    doi: 10.1126/sciadv.ads7509

    Figure Lengend Snippet: ( A ) eMAP workflow. ( B ) Representative images showing the dendrites of pyramidal neurons in the CA1 region of the 11-week-old mouse hippocampus. Arrow colors indicate different types of dendritic spines. Scale bars, 100 μm (left) and 10 μm (right), estimated to be 33.33- and 3.33-μm preexpansion, respectively. ( C and D ) Quantification of protrusion density (C), mushroom spine, stubby spine, thin spine, and filopodia density (D) at dendrites ( n = 22 to 29 dendrites from five mice per group; protrusion density, mushroom, and thin: Student’s t test, stubby and filopodia: Mann-Whitney test). ( E ) Representative images showing 3D reconstructions of dendrites. Scale bar, 10 μm, estimated to be 3.33-μm preexpansion. ( F and G ) Quantification (F) and distribution (G) of dendritic spine volume ( n = 699 to 716 dendritic spines from five mice per group; 25th, 50th, and 75th percentiles of the data are presented; Mann-Whitney test). ( H ) Representative images showing fluorescence images and 3D reconstructions of PSD95 puncta (red) at dendritic spines. Scale bar, 5 μm, estimated to be 1.67-μm preexpansion. ( I and J ) Quantification (I) and distribution (J) of PSD95 puncta volume at dendritic spines ( n = 280 to 304 PSD95 puncta from five mice per group; 25th, 50th, and 75th percentiles of the data are presented; Mann-Whitney test). ( K ) A representative image illustrating whole-cell patch clamp recordings from CA1 stratum radiatum neurons in acute hippocampal brain slices. ( L ) Representative traces of spontaneous excitatory postsynaptic currents (sEPSCs). ( M ) Quantification of the frequency sEPSCs ( n = 9 to 11 cells per group; average sEPSC frequency: Student’s t test, cumulative fraction: Kolmogorov-Smirnov test). Values in (C) and (D), (F) and (G), and (I) and (J) were normalized to 3× expansion factor. Data are mean ± SEM unless otherwise specified. * P < 0.05 and *** P < 0.001.

    Article Snippet: Volumetric analysis of dendritic spines and PSD95 puncta was performed using Imaris (version 10.0).

    Techniques: MANN-WHITNEY, Fluorescence, Patch Clamp

    ( A ) DiO-PSD95.FingR-Citrine was injected in barrel cortex of SST-Cre mice. ( B ) Schematic outlining injection and training timeline. Animals in fixed tissue analysis were time matched to control for viral expression time. ( C ) 10x image of SST-Cre mice expressing PSD95.FingR across the cortical column. Scale=100µm ( D ) 63x volumetric image of PSD95.FingR labeled somatostatin neurons. Scale=10µm ( E ) (Left) Zoomed image of PSD95.FingR puncta along a dendrite. (Right) Surface reconstruction of citrine signal. Scale=1µm ( F ) Immunohistochemical labeling of SST neurons (Left) PSD95.FingR-Citrine (Middle) calretinin expression (protein product of Calb2 gene) Scale=10µm (Right) merged channels. ( G ) Schematic indicating layer imaging field of view was captured (Layer 2/3). ( H ) Cumulative distribution of PSD95 puncta volume in layer 2/3 SST-O neurons. ( I ) Same as (H) for L2/3 SST-Calb2 neurons. ( J ) Within animal comparison of mean PSD95 puncta volume between SST-O (gray bars) and SST-Calb2 (pink bars) after experiencing acclimation (ACC), one, or five days of sensory association training (SAT). ( K-N ) Same as G-J but for L5 SST-O and SST-Calb2 SST neurons.

    Journal: bioRxiv

    Article Title: Long-lasting, subtype-specific regulation of somatostatin interneurons during sensory learning

    doi: 10.1101/2024.11.19.624383

    Figure Lengend Snippet: ( A ) DiO-PSD95.FingR-Citrine was injected in barrel cortex of SST-Cre mice. ( B ) Schematic outlining injection and training timeline. Animals in fixed tissue analysis were time matched to control for viral expression time. ( C ) 10x image of SST-Cre mice expressing PSD95.FingR across the cortical column. Scale=100µm ( D ) 63x volumetric image of PSD95.FingR labeled somatostatin neurons. Scale=10µm ( E ) (Left) Zoomed image of PSD95.FingR puncta along a dendrite. (Right) Surface reconstruction of citrine signal. Scale=1µm ( F ) Immunohistochemical labeling of SST neurons (Left) PSD95.FingR-Citrine (Middle) calretinin expression (protein product of Calb2 gene) Scale=10µm (Right) merged channels. ( G ) Schematic indicating layer imaging field of view was captured (Layer 2/3). ( H ) Cumulative distribution of PSD95 puncta volume in layer 2/3 SST-O neurons. ( I ) Same as (H) for L2/3 SST-Calb2 neurons. ( J ) Within animal comparison of mean PSD95 puncta volume between SST-O (gray bars) and SST-Calb2 (pink bars) after experiencing acclimation (ACC), one, or five days of sensory association training (SAT). ( K-N ) Same as G-J but for L5 SST-O and SST-Calb2 SST neurons.

    Article Snippet: For chemogenetic experiments designed to monitor changes in PSD95 puncta after suppressing activity, we coinjected pAAV8-hSyn-DIO-hM4Di-mCherry (Adgene #44362) with AAV-PHP.eB-ZFN-hSyn-DIO-PSD95.FingR-Citrine-reg.WPRE into S1BF using the stereotaxic injection procedures outlined above.

    Techniques: Injection, Control, Expressing, Labeling, Immunohistochemical staining, Imaging, Comparison

    ( A ) Region of interest (ROI) containing raw PSD95.FingR signal in confocal image stack (63x). Scale=5µm ( B ) Background subtraction prior surface reconstructions. ( C ) Semi-automated thresholding of 3D surface masks (gray) to optimize signal coverage while minimizing noise contained in background voxels. ( D ) A quality filter designating seed points (punctate signal) for splitting fused 3D surface masks. ( E ) 3D reconstructed fluorescence signal (multi-colored) after filtering structures to be ≥ 3 voxels to visualize all PSD95.FingR surface objects. ( F ) 3D surface mask of somatic PSD95.FingR signal. ( G ) 3D surface masks of PSD95.FingR signal within 0.15 um from reconstructed soma were removed. Remaining punctate PSD95.FingR signal (multi-colored) were included for whole field puncta analysis. ( H ) Zoomed out ROI from the same confocal image stack containing punctate PSD95.FingR signal for single cell puncta localization and subsequent comparisons between SST subtypes. Scale=15µm ( I ) Manually selected 3D surfaces (multi-colored) belonging to traceable dendrites emanating from soma in the top left corner. ( J ) Same as (I) but for soma in the bottom right corner. ( K ) Calb2-IR (purple) revealed for classifying puncta belonging to SST-Calb2 or SST-O. ( L ) Designated puncta belonging to SST-O (yellow) and SST-Calb2 (purple) neurons

    Journal: bioRxiv

    Article Title: Long-lasting, subtype-specific regulation of somatostatin interneurons during sensory learning

    doi: 10.1101/2024.11.19.624383

    Figure Lengend Snippet: ( A ) Region of interest (ROI) containing raw PSD95.FingR signal in confocal image stack (63x). Scale=5µm ( B ) Background subtraction prior surface reconstructions. ( C ) Semi-automated thresholding of 3D surface masks (gray) to optimize signal coverage while minimizing noise contained in background voxels. ( D ) A quality filter designating seed points (punctate signal) for splitting fused 3D surface masks. ( E ) 3D reconstructed fluorescence signal (multi-colored) after filtering structures to be ≥ 3 voxels to visualize all PSD95.FingR surface objects. ( F ) 3D surface mask of somatic PSD95.FingR signal. ( G ) 3D surface masks of PSD95.FingR signal within 0.15 um from reconstructed soma were removed. Remaining punctate PSD95.FingR signal (multi-colored) were included for whole field puncta analysis. ( H ) Zoomed out ROI from the same confocal image stack containing punctate PSD95.FingR signal for single cell puncta localization and subsequent comparisons between SST subtypes. Scale=15µm ( I ) Manually selected 3D surfaces (multi-colored) belonging to traceable dendrites emanating from soma in the top left corner. ( J ) Same as (I) but for soma in the bottom right corner. ( K ) Calb2-IR (purple) revealed for classifying puncta belonging to SST-Calb2 or SST-O. ( L ) Designated puncta belonging to SST-O (yellow) and SST-Calb2 (purple) neurons

    Article Snippet: For chemogenetic experiments designed to monitor changes in PSD95 puncta after suppressing activity, we coinjected pAAV8-hSyn-DIO-hM4Di-mCherry (Adgene #44362) with AAV-PHP.eB-ZFN-hSyn-DIO-PSD95.FingR-Citrine-reg.WPRE into S1BF using the stereotaxic injection procedures outlined above.

    Techniques: Fluorescence

    ( A ) Representative field of view from PSD95.FingR labeled SST neurons in L2/3 of primary somatosensory cortex. Scale=20µm. ( B ) Cumulative distribution of reconstructed PSD95.FingR surface volume from whole field analysis in acclimated (black dashed line), one day (thin blue line), and five day trained (thick blue line) SST-Cre mice. ( C ) Mean puncta volume in individual animals across training conditions. (ACC N=11 mice, 60,500 puncta; SAT1 N=8 mice, 44,000 puncta; SAT5 N=9 mice, 49,500) puncta ( D-F ) Same as A-C but for L4 PSD95.FingR labeled SST neurons. (ACC N=11 mice, 79,420 puncta; SAT1 N=8 mice, 57,760 puncta; SAT5 N=9 mice, 64,980 puncta; ACC vs SAT1 p=.04, unpaired t-test) ( G-I ) Same as A-C but for L5 PSD95.FingR labeled SST neurons. (ACC N=11 mice, 98,900 puncta; SAT1 N=8 mice, 79,120 puncta; SAT5 N=9 mice, 89,010 puncta).

    Journal: bioRxiv

    Article Title: Long-lasting, subtype-specific regulation of somatostatin interneurons during sensory learning

    doi: 10.1101/2024.11.19.624383

    Figure Lengend Snippet: ( A ) Representative field of view from PSD95.FingR labeled SST neurons in L2/3 of primary somatosensory cortex. Scale=20µm. ( B ) Cumulative distribution of reconstructed PSD95.FingR surface volume from whole field analysis in acclimated (black dashed line), one day (thin blue line), and five day trained (thick blue line) SST-Cre mice. ( C ) Mean puncta volume in individual animals across training conditions. (ACC N=11 mice, 60,500 puncta; SAT1 N=8 mice, 44,000 puncta; SAT5 N=9 mice, 49,500) puncta ( D-F ) Same as A-C but for L4 PSD95.FingR labeled SST neurons. (ACC N=11 mice, 79,420 puncta; SAT1 N=8 mice, 57,760 puncta; SAT5 N=9 mice, 64,980 puncta; ACC vs SAT1 p=.04, unpaired t-test) ( G-I ) Same as A-C but for L5 PSD95.FingR labeled SST neurons. (ACC N=11 mice, 98,900 puncta; SAT1 N=8 mice, 79,120 puncta; SAT5 N=9 mice, 89,010 puncta).

    Article Snippet: For chemogenetic experiments designed to monitor changes in PSD95 puncta after suppressing activity, we coinjected pAAV8-hSyn-DIO-hM4Di-mCherry (Adgene #44362) with AAV-PHP.eB-ZFN-hSyn-DIO-PSD95.FingR-Citrine-reg.WPRE into S1BF using the stereotaxic injection procedures outlined above.

    Techniques: Labeling

    ( A ) Schematic outlining viral injection strategy. ( B ) Schematic outlining training timeline. ( C ) 10x image of PSD95.FingR labeled SST neurons merged with calretinin-IR. Scale = 100µm ( D ) 63x confocal image stack containing PSD95.FingR labeled SST neurons merged with calretinin-IR. Scale = 20µm ( E ) Zoomed image of SST-O neuron. PSD95.FingR-Citrine (left), calretinin-IR (middle), overlay, (right). Scale = 5µm ( F ) Same as (E) but for a SST-Calb2 neuron. ( G ) Schematic outlining stimulus and reward presentation probabilities after trial initiations in pseudo-training. ( H ) Within animal comparison between SST-O neurons (gray bars) and SST-Calb2 (pink bars) during acclimation or 5 days of pseudo-training (PSE5). (ACC N = 4mice, 800 puncta; PSE N = 5 mice, 1000 puncta). ( I ) Image showing naïve (left) and environmentally enriched cages (right). ( J ) Same as (H) but for naïve animals and animals experiencing five days of environmental enrichment (EE5). (Naive N = 5mice, 1000 puncta; EE5 N = 5 mice, 1000 puncta)

    Journal: bioRxiv

    Article Title: Long-lasting, subtype-specific regulation of somatostatin interneurons during sensory learning

    doi: 10.1101/2024.11.19.624383

    Figure Lengend Snippet: ( A ) Schematic outlining viral injection strategy. ( B ) Schematic outlining training timeline. ( C ) 10x image of PSD95.FingR labeled SST neurons merged with calretinin-IR. Scale = 100µm ( D ) 63x confocal image stack containing PSD95.FingR labeled SST neurons merged with calretinin-IR. Scale = 20µm ( E ) Zoomed image of SST-O neuron. PSD95.FingR-Citrine (left), calretinin-IR (middle), overlay, (right). Scale = 5µm ( F ) Same as (E) but for a SST-Calb2 neuron. ( G ) Schematic outlining stimulus and reward presentation probabilities after trial initiations in pseudo-training. ( H ) Within animal comparison between SST-O neurons (gray bars) and SST-Calb2 (pink bars) during acclimation or 5 days of pseudo-training (PSE5). (ACC N = 4mice, 800 puncta; PSE N = 5 mice, 1000 puncta). ( I ) Image showing naïve (left) and environmentally enriched cages (right). ( J ) Same as (H) but for naïve animals and animals experiencing five days of environmental enrichment (EE5). (Naive N = 5mice, 1000 puncta; EE5 N = 5 mice, 1000 puncta)

    Article Snippet: For chemogenetic experiments designed to monitor changes in PSD95 puncta after suppressing activity, we coinjected pAAV8-hSyn-DIO-hM4Di-mCherry (Adgene #44362) with AAV-PHP.eB-ZFN-hSyn-DIO-PSD95.FingR-Citrine-reg.WPRE into S1BF using the stereotaxic injection procedures outlined above.

    Techniques: Injection, Labeling, Comparison

    Chemogenetic suppression of SST neuron activity reduces PSD95 puncta size on both SST-O and SST-Calb2 neurons . ( A ) Schematic of Viral injection. ( B ) Schematic of experimental timeline. ( C ) 63x somatic ROI of volumetric confocal image stack containing from left to right, PSD95.FingR-Citrine (yellow), calretinin-IR (purple), and hM4Di-mCherry (pseudo-colored cyan) fluorescence channels. ( D ) Dendrite example from an SST-O neuron (left) overlay between PSD95.FingR (yellow) and calretinin-IR (purple). (right) overlay between PSD95.FingR and hM4Di-mCherry (cyan). ( E, F ) same as in (C, D) but for an SST-O neuron transduced with hM4Di-mCherry. ( G ) Cumulative distribution of PSD95 puncta volume in SST-O neurons with and without hM4Di expression (hM4Di-N=6 mice, 1200 puncta; hM4Di+ N=6 mice, 1200 puncta). ( H ) Within animal comparison of average PSD95 puncta size in SST-O cells with and without hM4Di expression. ( I, J ) same as in (C, D) but an SST-Calb2 neuron not transduced with hM4Di-mCherry. ( K, L ) same as in (C, D) but for an SST-Calb2 neuron transduced with hM4Di-mCherry. ( M, N ) Same as in (G,H) but for SST-Calb2 neurons (hM4Di-N=6 mice, 1200 puncta; hM4Di+ N=6 mice, 1200 puncta).

    Journal: bioRxiv

    Article Title: Long-lasting, subtype-specific regulation of somatostatin interneurons during sensory learning

    doi: 10.1101/2024.11.19.624383

    Figure Lengend Snippet: Chemogenetic suppression of SST neuron activity reduces PSD95 puncta size on both SST-O and SST-Calb2 neurons . ( A ) Schematic of Viral injection. ( B ) Schematic of experimental timeline. ( C ) 63x somatic ROI of volumetric confocal image stack containing from left to right, PSD95.FingR-Citrine (yellow), calretinin-IR (purple), and hM4Di-mCherry (pseudo-colored cyan) fluorescence channels. ( D ) Dendrite example from an SST-O neuron (left) overlay between PSD95.FingR (yellow) and calretinin-IR (purple). (right) overlay between PSD95.FingR and hM4Di-mCherry (cyan). ( E, F ) same as in (C, D) but for an SST-O neuron transduced with hM4Di-mCherry. ( G ) Cumulative distribution of PSD95 puncta volume in SST-O neurons with and without hM4Di expression (hM4Di-N=6 mice, 1200 puncta; hM4Di+ N=6 mice, 1200 puncta). ( H ) Within animal comparison of average PSD95 puncta size in SST-O cells with and without hM4Di expression. ( I, J ) same as in (C, D) but an SST-Calb2 neuron not transduced with hM4Di-mCherry. ( K, L ) same as in (C, D) but for an SST-Calb2 neuron transduced with hM4Di-mCherry. ( M, N ) Same as in (G,H) but for SST-Calb2 neurons (hM4Di-N=6 mice, 1200 puncta; hM4Di+ N=6 mice, 1200 puncta).

    Article Snippet: For chemogenetic experiments designed to monitor changes in PSD95 puncta after suppressing activity, we coinjected pAAV8-hSyn-DIO-hM4Di-mCherry (Adgene #44362) with AAV-PHP.eB-ZFN-hSyn-DIO-PSD95.FingR-Citrine-reg.WPRE into S1BF using the stereotaxic injection procedures outlined above.

    Techniques: Activity Assay, Injection, Fluorescence, Transduction, Expressing, Comparison