neurobiotin tracer  (Vector Laboratories)


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    Vector Laboratories neurobiotin tracer
    Neurobiotin Tracer, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    neurobiotin tracer  (Vector Laboratories)


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    Vector Laboratories neurobiotin tracer
    Neurobiotin Tracer, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    neurobiotin  (Vector Laboratories)


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    Vector Laboratories neurobiotin
    KEY RESOURCES TABLE
    Neurobiotin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Unified classification of mouse retinal ganglion cells using function, morphology, and gene expression"

    Article Title: Unified classification of mouse retinal ganglion cells using function, morphology, and gene expression

    Journal: Cell reports

    doi: 10.1016/j.celrep.2022.111040

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Plasmid Preparation, Software

    neurobiotin  (Vector Laboratories)


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    Vector Laboratories neurobiotin
    KEY RESOURCES TABLE
    Neurobiotin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Unified classification of mouse retinal ganglion cells using function, morphology, and gene expression"

    Article Title: Unified classification of mouse retinal ganglion cells using function, morphology, and gene expression

    Journal: Cell reports

    doi: 10.1016/j.celrep.2022.111040

    KEY RESOURCES TABLE
    Figure Legend Snippet: KEY RESOURCES TABLE

    Techniques Used: Recombinant, Plasmid Preparation, Software

    neurobiotin  (Vector Laboratories)


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    Vector Laboratories neurobiotin
    a & b . Experimental design of in vivo Nrxn3 deletions and rescues using stereotactic infections of the OB with AAVs expressing ΔCre (control), Cre, or Cre with additional AAVs expressing Nrxn3-LNS2 rescue constructs (a, schematic of stereotactic injections; b, representative fluorescence image of an OB section that was infected with AAVs expressing tdTomato fused to Cre, with subsequent patching of mitral cells that were filled with <t>neurobiotin</t> (blue)). Note that, as illustrated in b, the AAVs infect granule cells more efficiently than mitral cells. c . The minimal Nrxn3α-LNS2 rescue constructs are localized to synaptic layers in the OB after expression via AAVs, as visualized by immunocytochemistry for the N-terminal HA-epitope contained in the constructs (white, HA-Nrxn3α-LNS2 proteins; red, tdTomato). d & e . Conditional deletion of Nrxn3 and rescue with minimal Nrxn3α-LNS2 constructs does not alter inhibitory synapse numbers in vivo . Sections from mice infected as shown in A were analyzed by quantitative immunocytochemistry for the presynaptic marker synaptoporin (a.k.a. synaptophysin-2; light blue) that is specific for granule cell→mitral cell synapses in the OB (Bergmann et al., 1993), and for the postsynaptic inhibitory synapse marker gephyrin (green) (d, sample images; e, summary graphs of puncta densities). Data are means ± SEM; n’s (cells/experiments) indicated in the summary graph bars apply to all graphs in an experimental series. Statistical analyses using one-way ANOVA with Dunnett’s multiple comparison test failed to uncover significant differences. Note that puncta densities in e are plotted both as analyzed per animal (top) and per region-of-interest (bottom); statistical significance is observed for gephyrin staining but not the other parameters when regions-of-interest are used as n’s because the pseudo-replicates in this analysis boost statistical significance independent of the actual number of experiments.
    Neurobiotin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "A combinatorial code of neurexin-3 alternative splicing controls inhibitory synapses via a trans-synaptic dystroglycan signaling loop"

    Article Title: A combinatorial code of neurexin-3 alternative splicing controls inhibitory synapses via a trans-synaptic dystroglycan signaling loop

    Journal: bioRxiv

    doi: 10.1101/2022.05.09.491206

    a & b . Experimental design of in vivo Nrxn3 deletions and rescues using stereotactic infections of the OB with AAVs expressing ΔCre (control), Cre, or Cre with additional AAVs expressing Nrxn3-LNS2 rescue constructs (a, schematic of stereotactic injections; b, representative fluorescence image of an OB section that was infected with AAVs expressing tdTomato fused to Cre, with subsequent patching of mitral cells that were filled with neurobiotin (blue)). Note that, as illustrated in b, the AAVs infect granule cells more efficiently than mitral cells. c . The minimal Nrxn3α-LNS2 rescue constructs are localized to synaptic layers in the OB after expression via AAVs, as visualized by immunocytochemistry for the N-terminal HA-epitope contained in the constructs (white, HA-Nrxn3α-LNS2 proteins; red, tdTomato). d & e . Conditional deletion of Nrxn3 and rescue with minimal Nrxn3α-LNS2 constructs does not alter inhibitory synapse numbers in vivo . Sections from mice infected as shown in A were analyzed by quantitative immunocytochemistry for the presynaptic marker synaptoporin (a.k.a. synaptophysin-2; light blue) that is specific for granule cell→mitral cell synapses in the OB (Bergmann et al., 1993), and for the postsynaptic inhibitory synapse marker gephyrin (green) (d, sample images; e, summary graphs of puncta densities). Data are means ± SEM; n’s (cells/experiments) indicated in the summary graph bars apply to all graphs in an experimental series. Statistical analyses using one-way ANOVA with Dunnett’s multiple comparison test failed to uncover significant differences. Note that puncta densities in e are plotted both as analyzed per animal (top) and per region-of-interest (bottom); statistical significance is observed for gephyrin staining but not the other parameters when regions-of-interest are used as n’s because the pseudo-replicates in this analysis boost statistical significance independent of the actual number of experiments.
    Figure Legend Snippet: a & b . Experimental design of in vivo Nrxn3 deletions and rescues using stereotactic infections of the OB with AAVs expressing ΔCre (control), Cre, or Cre with additional AAVs expressing Nrxn3-LNS2 rescue constructs (a, schematic of stereotactic injections; b, representative fluorescence image of an OB section that was infected with AAVs expressing tdTomato fused to Cre, with subsequent patching of mitral cells that were filled with neurobiotin (blue)). Note that, as illustrated in b, the AAVs infect granule cells more efficiently than mitral cells. c . The minimal Nrxn3α-LNS2 rescue constructs are localized to synaptic layers in the OB after expression via AAVs, as visualized by immunocytochemistry for the N-terminal HA-epitope contained in the constructs (white, HA-Nrxn3α-LNS2 proteins; red, tdTomato). d & e . Conditional deletion of Nrxn3 and rescue with minimal Nrxn3α-LNS2 constructs does not alter inhibitory synapse numbers in vivo . Sections from mice infected as shown in A were analyzed by quantitative immunocytochemistry for the presynaptic marker synaptoporin (a.k.a. synaptophysin-2; light blue) that is specific for granule cell→mitral cell synapses in the OB (Bergmann et al., 1993), and for the postsynaptic inhibitory synapse marker gephyrin (green) (d, sample images; e, summary graphs of puncta densities). Data are means ± SEM; n’s (cells/experiments) indicated in the summary graph bars apply to all graphs in an experimental series. Statistical analyses using one-way ANOVA with Dunnett’s multiple comparison test failed to uncover significant differences. Note that puncta densities in e are plotted both as analyzed per animal (top) and per region-of-interest (bottom); statistical significance is observed for gephyrin staining but not the other parameters when regions-of-interest are used as n’s because the pseudo-replicates in this analysis boost statistical significance independent of the actual number of experiments.

    Techniques Used: In Vivo, Expressing, Construct, Fluorescence, Infection, Immunocytochemistry, Marker, Staining

    a. Design of in vivo Nrxn3 deletions using stereotactic infections of the mPFC with AAVs expressing ΔCre (control), Cre, or Cre with the minimal Nrxn3α-LNS2 rescue constructs. b. Representative fluorescence image of an mPFC section from a mouse infected with AAVs expressing Cre-tdTomato (red) in which a layer 5 pyramidal neuron was patched and filled with neurobiotin (expanded right image; blue). c - e . The Nrxn3 deletion decreases the mIPSC frequency in vivo ; this decrease is rescued by the minimal Nrxn3α-LNS2 construct lacking an insert in SS2 (c, representative mIPSC traces recorded in the presence of TTX; d, cumulative probability plots of the interevent interval and summary graph of the mIPSC frequency; e, cumulative probability plots and summary graph of the mIPSC amplitudes). f - h . The Nrxn3 deletion greatly decreases the amplitude of IPSCs evoked by extracellular stimulation in layer 5 and recorded from pyramidal neurons in layer 5; again, this phenotype is rescued by the minimal Nrxn3α-LNS2 construct lacking an insert in SS2, as documented by input/output curves to control for possible variations in stimulating electrode placement (f, representative IPSC traces; g, summary plot of input/output amplitude measurements; h, summary graph of the slope of the input/output curves). Note that the input/output curves are not linear above 20 μA stimuli, suggesting a saturable number of axons from inhibitory neurons that form synapses on layer 5 neurons. i . The Nrxn3 deletion increases the rise time of evoked IPSCs (left) in a manner that can be rescued by the minimal Nrxn3α-LNS2 rescue constructs. In contrast, Nrxn3 deletion and rescue construct expression have no effect on decay times (right) of evoked IPSCs. j . The Nrxn3 deletion causes a substantial elevation in the coefficient of variation of IPSCs, suggesting a decrease in release probability, in a manner that can be rescued by the minimal Nrxn3α-LNS2 construct lacking an insert in SS2. Numerical data are means ± SEM; n’s (cells/experiments) are indicated above the sample traces and apply to all graphs in an experimental series. Statistical analyses were performed using two-way ANOVA in g and one-way ANOVA in d, e, h-j with Dunnett’s multiple comparison test with regards to the ΔCre group, with * = p<0.05, ** = p<0.01, and *** = p<0.0001.
    Figure Legend Snippet: a. Design of in vivo Nrxn3 deletions using stereotactic infections of the mPFC with AAVs expressing ΔCre (control), Cre, or Cre with the minimal Nrxn3α-LNS2 rescue constructs. b. Representative fluorescence image of an mPFC section from a mouse infected with AAVs expressing Cre-tdTomato (red) in which a layer 5 pyramidal neuron was patched and filled with neurobiotin (expanded right image; blue). c - e . The Nrxn3 deletion decreases the mIPSC frequency in vivo ; this decrease is rescued by the minimal Nrxn3α-LNS2 construct lacking an insert in SS2 (c, representative mIPSC traces recorded in the presence of TTX; d, cumulative probability plots of the interevent interval and summary graph of the mIPSC frequency; e, cumulative probability plots and summary graph of the mIPSC amplitudes). f - h . The Nrxn3 deletion greatly decreases the amplitude of IPSCs evoked by extracellular stimulation in layer 5 and recorded from pyramidal neurons in layer 5; again, this phenotype is rescued by the minimal Nrxn3α-LNS2 construct lacking an insert in SS2, as documented by input/output curves to control for possible variations in stimulating electrode placement (f, representative IPSC traces; g, summary plot of input/output amplitude measurements; h, summary graph of the slope of the input/output curves). Note that the input/output curves are not linear above 20 μA stimuli, suggesting a saturable number of axons from inhibitory neurons that form synapses on layer 5 neurons. i . The Nrxn3 deletion increases the rise time of evoked IPSCs (left) in a manner that can be rescued by the minimal Nrxn3α-LNS2 rescue constructs. In contrast, Nrxn3 deletion and rescue construct expression have no effect on decay times (right) of evoked IPSCs. j . The Nrxn3 deletion causes a substantial elevation in the coefficient of variation of IPSCs, suggesting a decrease in release probability, in a manner that can be rescued by the minimal Nrxn3α-LNS2 construct lacking an insert in SS2. Numerical data are means ± SEM; n’s (cells/experiments) are indicated above the sample traces and apply to all graphs in an experimental series. Statistical analyses were performed using two-way ANOVA in g and one-way ANOVA in d, e, h-j with Dunnett’s multiple comparison test with regards to the ΔCre group, with * = p<0.05, ** = p<0.01, and *** = p<0.0001.

    Techniques Used: In Vivo, Expressing, Construct, Fluorescence, Infection

    a. Experimental design. The OB of constitutive CAS9-expressing mice was infected stereotactically with AAVs encoding control or Dag1 gRNAs together with tdTomato at P15-18, and mice were analyzed 2-3 weeks later. b. Representative fluorescence images of OB sections stained for gephyrin as an inhibitory synapse marker (green) and tdTomato expressed by the AAVs (red). c & d . Dystroglycan ( Dag1 ) deletion in the OB in vivo does not change the density or size of gephyrin-positive synaptic puncta. Sections from mice infected as shown in a and b were analyzed by quantitative immunocytochemistry for the postsynaptic inhibitory synapse marker gephyrin (green) (c, sample images; d, summary graphs of puncta densities (top) and size (bottom)). Puncta densities and sizes are plotted as analyzed per animal, not per region-of-interest. e . Representative fluorescence image of a mitral cell filled with neurobiotin (blue) via the patch pipette in an OB section from a mouse with CRISPR-induced dystroglycan deletion in the OB (tdTomato expressed by the AAVs is shown in red, and EGFP expressed via the CAS9 knockin in green). f - h . Dystroglycan deletion in the OB in vivo decreases the mIPSC frequency monitored in mitral cells (f, representative mIPSC traces recorded in the presence of TTX; g, cumulative probability plots of the interevent interval and summary graph of the mIPSC frequency; h, cumulative probability plots and summary graph of the mIPSC amplitudes). i - k . Dystroglycan deletion in the OB in vivo suppresses inhibitory GC→MC synaptic transmission evoked by extracellular stimulation, as documented by input/output curves to control for variations in stimulating electrode placement (i, representative IPSC traces; j, summary plot of input/output amplitude measurements; k, summary graph of the slope of the input/output curves). l . Dystroglycan deletion in the OB in vivo causes a massive increase in the coefficient of variation of evoked IPSCs at GC→MC synapses, suggesting a decrease in release probability. m & n . Consistent with a decreased release probability, the Dag1 deletion induces a large increase in the paired-pulse ratio (m, representative traces; n, summary plot of the paired-pulse ratio). Numerical data are means ± SEM; n’s (cells/experiments) are indicated in the summary graph bars (d) or above the sample traces (f, i and m) and apply to all graphs in an experimental series. Statistical analyses were performed using Student’s t-test in d, g, h, k, l and two-way ANOVA in j & n, with * = p<0.05, ** = p<0.01, *** = p<0.001, and **** = p<0.0001.
    Figure Legend Snippet: a. Experimental design. The OB of constitutive CAS9-expressing mice was infected stereotactically with AAVs encoding control or Dag1 gRNAs together with tdTomato at P15-18, and mice were analyzed 2-3 weeks later. b. Representative fluorescence images of OB sections stained for gephyrin as an inhibitory synapse marker (green) and tdTomato expressed by the AAVs (red). c & d . Dystroglycan ( Dag1 ) deletion in the OB in vivo does not change the density or size of gephyrin-positive synaptic puncta. Sections from mice infected as shown in a and b were analyzed by quantitative immunocytochemistry for the postsynaptic inhibitory synapse marker gephyrin (green) (c, sample images; d, summary graphs of puncta densities (top) and size (bottom)). Puncta densities and sizes are plotted as analyzed per animal, not per region-of-interest. e . Representative fluorescence image of a mitral cell filled with neurobiotin (blue) via the patch pipette in an OB section from a mouse with CRISPR-induced dystroglycan deletion in the OB (tdTomato expressed by the AAVs is shown in red, and EGFP expressed via the CAS9 knockin in green). f - h . Dystroglycan deletion in the OB in vivo decreases the mIPSC frequency monitored in mitral cells (f, representative mIPSC traces recorded in the presence of TTX; g, cumulative probability plots of the interevent interval and summary graph of the mIPSC frequency; h, cumulative probability plots and summary graph of the mIPSC amplitudes). i - k . Dystroglycan deletion in the OB in vivo suppresses inhibitory GC→MC synaptic transmission evoked by extracellular stimulation, as documented by input/output curves to control for variations in stimulating electrode placement (i, representative IPSC traces; j, summary plot of input/output amplitude measurements; k, summary graph of the slope of the input/output curves). l . Dystroglycan deletion in the OB in vivo causes a massive increase in the coefficient of variation of evoked IPSCs at GC→MC synapses, suggesting a decrease in release probability. m & n . Consistent with a decreased release probability, the Dag1 deletion induces a large increase in the paired-pulse ratio (m, representative traces; n, summary plot of the paired-pulse ratio). Numerical data are means ± SEM; n’s (cells/experiments) are indicated in the summary graph bars (d) or above the sample traces (f, i and m) and apply to all graphs in an experimental series. Statistical analyses were performed using Student’s t-test in d, g, h, k, l and two-way ANOVA in j & n, with * = p<0.05, ** = p<0.01, *** = p<0.001, and **** = p<0.0001.

    Techniques Used: Expressing, Infection, Fluorescence, Staining, Marker, In Vivo, Immunocytochemistry, Transferring, CRISPR, Knock-In, Transmission Assay

    a. Experimental design for mitral cell-specific dystroglycan deletions in the OB using retro-AAVs encoding a control gRNA or the dystroglycan ( Dag1 ) gRNA together with tdTomato that are injected into the piriform cortex of constitutively CAS9 expressing mice. The retro-AAVs infect axons projecting from OB mitral/tufted cells and thereby delete dystroglycan specifically in mitral/tufted cells of the OB. b. Representative fluorescence images of OB sections stained for gephyrin as an inhibitory synapse marker (green) and tdTomato expressed by the retro-AAVs (red). c & d . Dystroglycan deletion in vivo does not change the density or size of gephyrin-positive synaptic puncta in the OB. Sections from mice infected as shown in a and b were analyzed by quantitative immunocytochemistry for the postsynaptic inhibitory synapse marker gephyrin (green) (c, sample images; d, summary graphs of puncta densities (top) and size (bottom)). Only gephyrin puncta that co-localized with tdTomato-expressing mitral cell somata and dendritic segments were quantified. Puncta densities and sizes are plotted as analyzed per animal, not per region-of-interest. e . Representative fluorescence image of a mitral cell filled with neurobiotin (blue) via the patch pipette in an OB section from a mouse with CRISPR-induced dystroglycan ( Dag1 ) deletion in mitral cells (tdTomato expressed by the retro-AAVs is shown in red). f - h . The dystroglycan ( Dag1 ) deletion decreases the mIPSC frequency in vivo in mitral cells (f, representative mIPSC traces recorded in the presence of TTX; g, cumulative probability plots of the interevent interval and summary graph of the mIPSC frequency; h, cumulative probability plots and summary graph of the mIPSC amplitudes). i - k . The dystroglycan ( Dag1 ) deletion greatly decreases the amplitude of IPSCs at GC→MC synapses evoked by extracellular stimulation, as documented by input/output curves to control for possible variations in stimulating electrode placement (i, representative IPSC traces; j, summary plot of input/output amplitude measurements; k, summary graph of the slope of the input/output curves). l . The mitral cell-specific dystroglycan ( Dag1 ) deletion causes a massive increase in the coefficient of variation of evoked IPSCs at GC→MC synapses, suggesting a decrease in release probability. m & n . Consistent with a decreased release probability, the dystroglycan ( Dag1 ) deletion induces a large increase in the paired-pulse ratio (m, representative traces; n, summary plot of the paired-pulse ratio). o - r . Dystroglycan deletion in the mPFC in vivo decreases the mIPSC frequency monitored in Layer 5 pyramidal neurons (o, representative image of an mPFC section; p, representative mIPSC traces recorded in the presence of TTX; q, cumulative probability plots of the interevent interval and summary graph of the mIPSC frequency; r, cumulative probability plots and summary graph of the mIPSC amplitudes). Numerical data are means ± SEM; n’s (cells/experiments) are indicated in the summary graph bars (d) or above the sample traces (f, i, m, and p, and apply to all graphs in an experimental series. Statistical analyses were performed using Student’s t-test in d, g, h, k, l, q, r, and by two-way ANOVA in j & n, with * = p<0.05, ** = p<0.01, *** = p<0.001, and *** = p<0.0001.
    Figure Legend Snippet: a. Experimental design for mitral cell-specific dystroglycan deletions in the OB using retro-AAVs encoding a control gRNA or the dystroglycan ( Dag1 ) gRNA together with tdTomato that are injected into the piriform cortex of constitutively CAS9 expressing mice. The retro-AAVs infect axons projecting from OB mitral/tufted cells and thereby delete dystroglycan specifically in mitral/tufted cells of the OB. b. Representative fluorescence images of OB sections stained for gephyrin as an inhibitory synapse marker (green) and tdTomato expressed by the retro-AAVs (red). c & d . Dystroglycan deletion in vivo does not change the density or size of gephyrin-positive synaptic puncta in the OB. Sections from mice infected as shown in a and b were analyzed by quantitative immunocytochemistry for the postsynaptic inhibitory synapse marker gephyrin (green) (c, sample images; d, summary graphs of puncta densities (top) and size (bottom)). Only gephyrin puncta that co-localized with tdTomato-expressing mitral cell somata and dendritic segments were quantified. Puncta densities and sizes are plotted as analyzed per animal, not per region-of-interest. e . Representative fluorescence image of a mitral cell filled with neurobiotin (blue) via the patch pipette in an OB section from a mouse with CRISPR-induced dystroglycan ( Dag1 ) deletion in mitral cells (tdTomato expressed by the retro-AAVs is shown in red). f - h . The dystroglycan ( Dag1 ) deletion decreases the mIPSC frequency in vivo in mitral cells (f, representative mIPSC traces recorded in the presence of TTX; g, cumulative probability plots of the interevent interval and summary graph of the mIPSC frequency; h, cumulative probability plots and summary graph of the mIPSC amplitudes). i - k . The dystroglycan ( Dag1 ) deletion greatly decreases the amplitude of IPSCs at GC→MC synapses evoked by extracellular stimulation, as documented by input/output curves to control for possible variations in stimulating electrode placement (i, representative IPSC traces; j, summary plot of input/output amplitude measurements; k, summary graph of the slope of the input/output curves). l . The mitral cell-specific dystroglycan ( Dag1 ) deletion causes a massive increase in the coefficient of variation of evoked IPSCs at GC→MC synapses, suggesting a decrease in release probability. m & n . Consistent with a decreased release probability, the dystroglycan ( Dag1 ) deletion induces a large increase in the paired-pulse ratio (m, representative traces; n, summary plot of the paired-pulse ratio). o - r . Dystroglycan deletion in the mPFC in vivo decreases the mIPSC frequency monitored in Layer 5 pyramidal neurons (o, representative image of an mPFC section; p, representative mIPSC traces recorded in the presence of TTX; q, cumulative probability plots of the interevent interval and summary graph of the mIPSC frequency; r, cumulative probability plots and summary graph of the mIPSC amplitudes). Numerical data are means ± SEM; n’s (cells/experiments) are indicated in the summary graph bars (d) or above the sample traces (f, i, m, and p, and apply to all graphs in an experimental series. Statistical analyses were performed using Student’s t-test in d, g, h, k, l, q, r, and by two-way ANOVA in j & n, with * = p<0.05, ** = p<0.01, *** = p<0.001, and *** = p<0.0001.

    Techniques Used: Injection, Expressing, Fluorescence, Staining, Marker, In Vivo, Infection, Immunocytochemistry, Transferring, CRISPR

    neurobiotin  (Vector Laboratories)


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    Vector Laboratories neurobiotin
    Neurobiotin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    neurobiotin  (Vector Laboratories)


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    Vector Laboratories neurobiotin
    Neurobiotin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    neurobiotin  (Vector Laboratories)


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    Vector Laboratories neurobiotin
    Identified CA2 Pyr neurons. A–E, Epifluorescence pictures showing PCP4 staining (green) and juxtacellularly labeled CA2 Pyr neurons (red), recorded in freely-moving mice (dataset related to Fig. 4M,N). Left panels, Low-magnification pictures (Pyr layer outlined as dotted lines). Right panels, High-magnification pictures, showing the location of the Pyr cell somata within CA2. A, Two neurons were sequentially labeled on the same electrode penetration: one FS cell (empty arrowhead) and one Pyr neuron (full arrowhead). The FS interneuron was negative for PCP4 expression, in line with previous observations (Kohara et al., 2014). B, Two CA2 Pyr neurons were labeled: one cell on a medial electrode penetration (dendrites indicated by the empty arrowhead), and another one on a more lateral penetration (full arrowhead). GCL, Granule cell layer; Nb, <t>neurobiotin.</t> Scale bars: A, 200 µm, Inset, 25 µm; B–E, 200 µm, Insets, 50 µm.
    Neurobiotin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Structural Correlates of CA2 and CA3 Pyramidal Cell Activity in Freely-Moving Mice"

    Article Title: Structural Correlates of CA2 and CA3 Pyramidal Cell Activity in Freely-Moving Mice

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.0099-20.2020

    Identified CA2 Pyr neurons. A–E, Epifluorescence pictures showing PCP4 staining (green) and juxtacellularly labeled CA2 Pyr neurons (red), recorded in freely-moving mice (dataset related to Fig. 4M,N). Left panels, Low-magnification pictures (Pyr layer outlined as dotted lines). Right panels, High-magnification pictures, showing the location of the Pyr cell somata within CA2. A, Two neurons were sequentially labeled on the same electrode penetration: one FS cell (empty arrowhead) and one Pyr neuron (full arrowhead). The FS interneuron was negative for PCP4 expression, in line with previous observations (Kohara et al., 2014). B, Two CA2 Pyr neurons were labeled: one cell on a medial electrode penetration (dendrites indicated by the empty arrowhead), and another one on a more lateral penetration (full arrowhead). GCL, Granule cell layer; Nb, neurobiotin. Scale bars: A, 200 µm, Inset, 25 µm; B–E, 200 µm, Insets, 50 µm.
    Figure Legend Snippet: Identified CA2 Pyr neurons. A–E, Epifluorescence pictures showing PCP4 staining (green) and juxtacellularly labeled CA2 Pyr neurons (red), recorded in freely-moving mice (dataset related to Fig. 4M,N). Left panels, Low-magnification pictures (Pyr layer outlined as dotted lines). Right panels, High-magnification pictures, showing the location of the Pyr cell somata within CA2. A, Two neurons were sequentially labeled on the same electrode penetration: one FS cell (empty arrowhead) and one Pyr neuron (full arrowhead). The FS interneuron was negative for PCP4 expression, in line with previous observations (Kohara et al., 2014). B, Two CA2 Pyr neurons were labeled: one cell on a medial electrode penetration (dendrites indicated by the empty arrowhead), and another one on a more lateral penetration (full arrowhead). GCL, Granule cell layer; Nb, neurobiotin. Scale bars: A, 200 µm, Inset, 25 µm; B–E, 200 µm, Insets, 50 µm.

    Techniques Used: Staining, Labeling, Expressing

    Identified PCP4-positive CA3a neurons. A, Epifluorescence pictures showing PCP4 staining (green) and a PCP4-positive CA3a Pyr neuron (Nb, neurobiotin, in red; see arrowhead), recorded in freely-moving mice (indicated by 1 in Fig. 4M,N). Bottom left, High-magnification view of the Pyr cell soma. Scale bars: 100 µm; Inset, 20 µm. B, Maximum intensity z-stack projections of the neuron in A after DAB conversion (see Materials and Methods), showing the soma (left) and proximal apical dendritic segments (right). Note the absence of thorny excrescences. Scale bars, 15 µm. C, Reconstruction of the dendritic morphology of the neuron shown in A, B. Scale bar, 50 µm. D–F, Same as in A–C, but for another PCP4-positive Pyr located in CA3a (indicated by 2 in Fig. 4M,N). Scale bars, same as in A–C. G, Maximum intensity z-stack projections of two representative CA3 cells, showing the presence of thorny excrescences on proximal dendritic segments (some are indicated by white arrowheads). Scale bars, 15 µm.
    Figure Legend Snippet: Identified PCP4-positive CA3a neurons. A, Epifluorescence pictures showing PCP4 staining (green) and a PCP4-positive CA3a Pyr neuron (Nb, neurobiotin, in red; see arrowhead), recorded in freely-moving mice (indicated by 1 in Fig. 4M,N). Bottom left, High-magnification view of the Pyr cell soma. Scale bars: 100 µm; Inset, 20 µm. B, Maximum intensity z-stack projections of the neuron in A after DAB conversion (see Materials and Methods), showing the soma (left) and proximal apical dendritic segments (right). Note the absence of thorny excrescences. Scale bars, 15 µm. C, Reconstruction of the dendritic morphology of the neuron shown in A, B. Scale bar, 50 µm. D–F, Same as in A–C, but for another PCP4-positive Pyr located in CA3a (indicated by 2 in Fig. 4M,N). Scale bars, same as in A–C. G, Maximum intensity z-stack projections of two representative CA3 cells, showing the presence of thorny excrescences on proximal dendritic segments (some are indicated by white arrowheads). Scale bars, 15 µm.

    Techniques Used: Staining

    neurobiotin  (Vector Laboratories)


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    Vector Laboratories neurobiotin
    Neurobiotin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    neurobiotin  (Vector Laboratories)


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    Vector Laboratories neurobiotin
    Images of the ganglion cell layer in a patch of retina in which a single F-mini-ON RGC was filled with <t>Neurobiotin</t> (magenta arrowhead). Left panel shows the Neurobiotin channel, with three brightly labelled coupled cells (white arrowheads) and three dimly labelled cells that likely represent second-order connections (magenta asterisks). Middle panel shows the same region with immunoreactivity for FOXP1, which labels F-mini-OFF RGCs, but does not label F-mini-ON RGCs . Right panel shows immunoreactivity for FOXP2, which labels both F-mini RGC types. This experiment was performed on five F-mini RGC networks in four retinas: four F-mini-ON RGCs and one F-mini-OFF RGC injected. Three networks were stained for FOXP2 and FOXP1; two networks for FoOXP2 only. Neurobiotin labeled 9.0 ± 6.4 somas per retina, and was found in varying amounts in neurons; indicating first and second order connectivity. FOXP2 was present in 43 of 45 RGCs that were labeled with Neurobiotin. Coupled cells from these networks that could be morphologically identified by using the visible primary dendrites, and all showed the expected patterns of FoxP1 expression. 8/8 F-mini-ON RGCs were FOXP1 negative and 14/14 F-mini-OFF RGCs were FOXP1 positive.
    Neurobiotin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "An offset ON-OFF receptive field is created by gap junctions between distinct types of retinal ganglion cells"

    Article Title: An offset ON-OFF receptive field is created by gap junctions between distinct types of retinal ganglion cells

    Journal: Nature neuroscience

    doi: 10.1038/s41593-020-00747-8

    Images of the ganglion cell layer in a patch of retina in which a single F-mini-ON RGC was filled with Neurobiotin (magenta arrowhead). Left panel shows the Neurobiotin channel, with three brightly labelled coupled cells (white arrowheads) and three dimly labelled cells that likely represent second-order connections (magenta asterisks). Middle panel shows the same region with immunoreactivity for FOXP1, which labels F-mini-OFF RGCs, but does not label F-mini-ON RGCs . Right panel shows immunoreactivity for FOXP2, which labels both F-mini RGC types. This experiment was performed on five F-mini RGC networks in four retinas: four F-mini-ON RGCs and one F-mini-OFF RGC injected. Three networks were stained for FOXP2 and FOXP1; two networks for FoOXP2 only. Neurobiotin labeled 9.0 ± 6.4 somas per retina, and was found in varying amounts in neurons; indicating first and second order connectivity. FOXP2 was present in 43 of 45 RGCs that were labeled with Neurobiotin. Coupled cells from these networks that could be morphologically identified by using the visible primary dendrites, and all showed the expected patterns of FoxP1 expression. 8/8 F-mini-ON RGCs were FOXP1 negative and 14/14 F-mini-OFF RGCs were FOXP1 positive.
    Figure Legend Snippet: Images of the ganglion cell layer in a patch of retina in which a single F-mini-ON RGC was filled with Neurobiotin (magenta arrowhead). Left panel shows the Neurobiotin channel, with three brightly labelled coupled cells (white arrowheads) and three dimly labelled cells that likely represent second-order connections (magenta asterisks). Middle panel shows the same region with immunoreactivity for FOXP1, which labels F-mini-OFF RGCs, but does not label F-mini-ON RGCs . Right panel shows immunoreactivity for FOXP2, which labels both F-mini RGC types. This experiment was performed on five F-mini RGC networks in four retinas: four F-mini-ON RGCs and one F-mini-OFF RGC injected. Three networks were stained for FOXP2 and FOXP1; two networks for FoOXP2 only. Neurobiotin labeled 9.0 ± 6.4 somas per retina, and was found in varying amounts in neurons; indicating first and second order connectivity. FOXP2 was present in 43 of 45 RGCs that were labeled with Neurobiotin. Coupled cells from these networks that could be morphologically identified by using the visible primary dendrites, and all showed the expected patterns of FoxP1 expression. 8/8 F-mini-ON RGCs were FOXP1 negative and 14/14 F-mini-OFF RGCs were FOXP1 positive.

    Techniques Used: Injection, Staining, Labeling, Expressing

    Image of the ganglion cell layer in a patch of retina in which a single F-mini-ON RGC was filled with Neurobiotin (arrow). RGC somas labeled by the dye are circled: cyan F-mini-OFF (first degree connections) and magenta F-mini-ON (filled RGC and second degree connections). The transcription factor FOXP2 is found in all F-mini RGCs (red); FOXP1 is found in F-mini-OFF RGCs (blue). Results were consistent over several images ( n = 5 filled networks on 4 retinas).
    Figure Legend Snippet: Image of the ganglion cell layer in a patch of retina in which a single F-mini-ON RGC was filled with Neurobiotin (arrow). RGC somas labeled by the dye are circled: cyan F-mini-OFF (first degree connections) and magenta F-mini-ON (filled RGC and second degree connections). The transcription factor FOXP2 is found in all F-mini RGCs (red); FOXP1 is found in F-mini-OFF RGCs (blue). Results were consistent over several images ( n = 5 filled networks on 4 retinas).

    Techniques Used: Labeling

    a, An example illustrating heterotypic network connectivity. The F-mini-OFF RGC labeled ‘0’ was filled with Alexa Fluor 488 (cyan), revealing 7 coupled somas (white). b, Cell-attached recordings from each of the labeled somas shown in a . Cells 1 through 6 show clear signs of being neighboring F-mini-ON RGCs. The soma of Cell 7 is dimmer, and is likely a second-order connected F-mini-OFF RGC. c, Distribution of the number of dye coupled cells observed in Neurobiotin and Alexa Fluor 488 cell fills of F-mini-ON and F-mini-OFF RGCs, n = 13, 3 cells. d, Average voltage traces from an RGC pair in which one F-mini RGC was injected with current (top row) and the coupled F-mini RGC of the opposite type (bottom row) showed a response. Current injections were +50 pA (lighter traces) and −50 pA (darker traces). e, Voltage change relationship across the electrical synapse in both directions for the pair in d . f, Distribution of the coupling coefficient (slope of line in e ) for all recorded pairs, in control conditions (top) and in the presence of MFA (bottom). g, Example of MFA abolishing voltage deflection, showing voltage in F-mini RGCs (for −50 pA injection in coupled cell) in control conditions and in the presence of MFA (green). Image in a is a composite of a maximum projection image of the F-mini-OFF dendrites in cyan with a maximum projection image of the ganglion cell layer in white. Cell ‘0’ in b was recorded in current clamp mode. Box plots in c show maximum, 75th percentile, median, 25th percentile, and minimum.
    Figure Legend Snippet: a, An example illustrating heterotypic network connectivity. The F-mini-OFF RGC labeled ‘0’ was filled with Alexa Fluor 488 (cyan), revealing 7 coupled somas (white). b, Cell-attached recordings from each of the labeled somas shown in a . Cells 1 through 6 show clear signs of being neighboring F-mini-ON RGCs. The soma of Cell 7 is dimmer, and is likely a second-order connected F-mini-OFF RGC. c, Distribution of the number of dye coupled cells observed in Neurobiotin and Alexa Fluor 488 cell fills of F-mini-ON and F-mini-OFF RGCs, n = 13, 3 cells. d, Average voltage traces from an RGC pair in which one F-mini RGC was injected with current (top row) and the coupled F-mini RGC of the opposite type (bottom row) showed a response. Current injections were +50 pA (lighter traces) and −50 pA (darker traces). e, Voltage change relationship across the electrical synapse in both directions for the pair in d . f, Distribution of the coupling coefficient (slope of line in e ) for all recorded pairs, in control conditions (top) and in the presence of MFA (bottom). g, Example of MFA abolishing voltage deflection, showing voltage in F-mini RGCs (for −50 pA injection in coupled cell) in control conditions and in the presence of MFA (green). Image in a is a composite of a maximum projection image of the F-mini-OFF dendrites in cyan with a maximum projection image of the ganglion cell layer in white. Cell ‘0’ in b was recorded in current clamp mode. Box plots in c show maximum, 75th percentile, median, 25th percentile, and minimum.

    Techniques Used: Labeling, Injection

    Three connexins were evaluated for presence at the regions of contact between an F-mini-ON and multiple F-mini-OFF RGCs, n = 1 of each experiment. a,b, Full depth maximum intensity projection images of a Neurobiotin-filled F-mini-ON RGC (magenta),the connected F-mini-OFF RGCs (cyan), and a cell of unclassified type due to insufficiently filled dendrites (yellow). Tracing, segmentation, and masking were performed manually. Image brightness was scaled separately by cell type for illustration here but not for analysis. c,d Thin projection images of regions in orange squares in a,b showing an example RGC crossing point with yellow square for spatial reference. Stack depth is 3.5 μm. e-g, The same region and depth as in c,d, showing the IHC channels for the three connexin proteins. h, Quantification of overlap between connexin images and RGC contact region masks. Values are similar before and after a 90 degree rotation of the connexin image. Points mark the overlap of the single F-mini-ON RGC with each F-mini-OFF RGC in the image.
    Figure Legend Snippet: Three connexins were evaluated for presence at the regions of contact between an F-mini-ON and multiple F-mini-OFF RGCs, n = 1 of each experiment. a,b, Full depth maximum intensity projection images of a Neurobiotin-filled F-mini-ON RGC (magenta),the connected F-mini-OFF RGCs (cyan), and a cell of unclassified type due to insufficiently filled dendrites (yellow). Tracing, segmentation, and masking were performed manually. Image brightness was scaled separately by cell type for illustration here but not for analysis. c,d Thin projection images of regions in orange squares in a,b showing an example RGC crossing point with yellow square for spatial reference. Stack depth is 3.5 μm. e-g, The same region and depth as in c,d, showing the IHC channels for the three connexin proteins. h, Quantification of overlap between connexin images and RGC contact region masks. Values are similar before and after a 90 degree rotation of the connexin image. Points mark the overlap of the single F-mini-ON RGC with each F-mini-OFF RGC in the image.

    Techniques Used:

    a, Traced microscope image from an experiment in which a single F-mini-ON RGC (magenta) was filled with Neurobiotin. Four coupled F-mini-OFF RGCs are in cyan. A spatial offset is apparent in the dendritic arbors of the two cell types relative to the soma of the F-mini-ON. b, Diagram of the imaging datasets used in the morphological model, described in d and e . c, RF model diagram (see ). Measurements of F-mini-ON coupled soma positions in d were randomly combined with convex polygon fits to the dendrites of F-mini-ON and F-mini-OFF RGCs in e , to create a purely anatomical prediction of the center-of-mass (COM) offset of ON and OFF RFs (red and blue crosses). d, Locations of labeled somas relative to the injected F-mini-ON RGCs (magenta circle) included both confirmed F-mini-OFF RGCs (squares) and RGC somas that were not further characterized (triangles) ( n = 50 somas, 13 injected cells, colored by injected cell) Each point represents the position of a gap-junction labeled soma relative to the position of the filled F-mini-ON RGC. Results above suggest that all coupled cells were in fact F-mini-OFF RGCs, but only some of them (squares) were confirmed via electrophysiology or IHC. e, Area of dendrites relative to soma position for the measured population of F-mini-ON and F-mini-OFF RGCs ( n = 38, 12 cells). Soma locations are marked by 50 μm crosses. f, Result from the model. Colored surface is mean OFF RF relative to the centered ON RF. True F-mini-ON RF offset data (black crosses) and format are from .
    Figure Legend Snippet: a, Traced microscope image from an experiment in which a single F-mini-ON RGC (magenta) was filled with Neurobiotin. Four coupled F-mini-OFF RGCs are in cyan. A spatial offset is apparent in the dendritic arbors of the two cell types relative to the soma of the F-mini-ON. b, Diagram of the imaging datasets used in the morphological model, described in d and e . c, RF model diagram (see ). Measurements of F-mini-ON coupled soma positions in d were randomly combined with convex polygon fits to the dendrites of F-mini-ON and F-mini-OFF RGCs in e , to create a purely anatomical prediction of the center-of-mass (COM) offset of ON and OFF RFs (red and blue crosses). d, Locations of labeled somas relative to the injected F-mini-ON RGCs (magenta circle) included both confirmed F-mini-OFF RGCs (squares) and RGC somas that were not further characterized (triangles) ( n = 50 somas, 13 injected cells, colored by injected cell) Each point represents the position of a gap-junction labeled soma relative to the position of the filled F-mini-ON RGC. Results above suggest that all coupled cells were in fact F-mini-OFF RGCs, but only some of them (squares) were confirmed via electrophysiology or IHC. e, Area of dendrites relative to soma position for the measured population of F-mini-ON and F-mini-OFF RGCs ( n = 38, 12 cells). Soma locations are marked by 50 μm crosses. f, Result from the model. Colored surface is mean OFF RF relative to the centered ON RF. True F-mini-ON RF offset data (black crosses) and format are from .

    Techniques Used: Microscopy, Imaging, Labeling, Injection

    neurobiotin  (Vector Laboratories)


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    Vector Laboratories neurobiotin tracer
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    KEY RESOURCES TABLE
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    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: Unified classification of mouse retinal ganglion cells using function, morphology, and gene expression

    doi: 10.1016/j.celrep.2022.111040

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: A subset of recorded RGCs were injected with Neurobiotin (Vector Laboratories, SP-1150, ~3% w/v and ~280 mOsm in potassium aspartate internal solution) using patch pipettes.

    Techniques: Recombinant, Plasmid Preparation, Software