rabbit α glt1  (Alomone Labs)


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    Alomone Labs rabbit α glt1
    <t>GLT1</t> HET mice have higher ipsilateral eye responses, lower contralateral eye bias and disrupted experience-dependent binocular matching of orientation-selective responses. A) Schematic of experimental design. Top: Visual gratings were separately presented to the contra (green) and ipsi (blue) eyes in P28 mice and neuronal responses recorded. Bottom: schematic of measures. Ocular dominance index (ODI) was calculated as (max Contra − max Ipsi ) / max Contra +max Ipsi . Orientation Selectivity Index (OSI) was calculated as described previously ( Banerjee et al., 2016 ). Difference in preferred orientation (ΔPO) was calculated as the difference between preferred orientations of the max contralateral and ipsilateral responses. B) Example cells in GLT1 WT (top) and GLT1 HET (bottom) animals. Left: in vivo images of neuronal somas measured in binocular visual cortex using the calcium indicator, GCaMP6f. Right: Tuning curves of three cells (white circles in left) to contra (green) and ipsi (blue) stimulation. Note the matched tuning and contralateral bias in WT animals and the mismatched tuning curves and lack of contralateral bias in GLT1 HETs. C) Quantification of the average response to PO in GLT1 WT and HET mice. WT mice have a significantly higher contralateral response than ipsilateral response while HET mice have approximately equal contralateral and ipsilateral responses (n=4-6 animals, 23-52 cells per animal, two-way ANOVA, genotype F(1,674)=7.72, p=0.0056, interaction F(1,674)=4.243, p=0.040). D) Quantification of ocular dominance index showing that GLT1 HET mice have significantly decreased ODI (n=4-6 animals, 23-52 cells per animal, t-test, p=0.0018). E) Quantification of OSI showing that GLT1 HET mice have a significantly decreased OSI of ipsilateral responses compared to both contra and ipsi responses in GLT1 WT animals (n=4-6 animals, 23-52 cells per animal, two-way ANOVA, genotype F(1,674)=12.46, p=4.5×10 −4 ). F) Quantification of ΔPO showing an increased difference in the preferred orientations between contralateral and ipsilateral inputs to neurons in GLT1 HET animals (n=4-6 animals, 23-52 cells per animal, t-test, p=1.0×10 −4 ). *p
    Rabbit α Glt1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Price from $9.99 to $1999.99
    rabbit α glt1 - by Bioz Stars, 2022-12
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    1) Product Images from "Astrocytic glutamate uptake coordinates experience-dependent, eye-specific refinement in developing visual cortex"

    Article Title: Astrocytic glutamate uptake coordinates experience-dependent, eye-specific refinement in developing visual cortex

    Journal: bioRxiv

    doi: 10.1101/2020.05.25.113613

    GLT1 HET mice have higher ipsilateral eye responses, lower contralateral eye bias and disrupted experience-dependent binocular matching of orientation-selective responses. A) Schematic of experimental design. Top: Visual gratings were separately presented to the contra (green) and ipsi (blue) eyes in P28 mice and neuronal responses recorded. Bottom: schematic of measures. Ocular dominance index (ODI) was calculated as (max Contra − max Ipsi ) / max Contra +max Ipsi . Orientation Selectivity Index (OSI) was calculated as described previously ( Banerjee et al., 2016 ). Difference in preferred orientation (ΔPO) was calculated as the difference between preferred orientations of the max contralateral and ipsilateral responses. B) Example cells in GLT1 WT (top) and GLT1 HET (bottom) animals. Left: in vivo images of neuronal somas measured in binocular visual cortex using the calcium indicator, GCaMP6f. Right: Tuning curves of three cells (white circles in left) to contra (green) and ipsi (blue) stimulation. Note the matched tuning and contralateral bias in WT animals and the mismatched tuning curves and lack of contralateral bias in GLT1 HETs. C) Quantification of the average response to PO in GLT1 WT and HET mice. WT mice have a significantly higher contralateral response than ipsilateral response while HET mice have approximately equal contralateral and ipsilateral responses (n=4-6 animals, 23-52 cells per animal, two-way ANOVA, genotype F(1,674)=7.72, p=0.0056, interaction F(1,674)=4.243, p=0.040). D) Quantification of ocular dominance index showing that GLT1 HET mice have significantly decreased ODI (n=4-6 animals, 23-52 cells per animal, t-test, p=0.0018). E) Quantification of OSI showing that GLT1 HET mice have a significantly decreased OSI of ipsilateral responses compared to both contra and ipsi responses in GLT1 WT animals (n=4-6 animals, 23-52 cells per animal, two-way ANOVA, genotype F(1,674)=12.46, p=4.5×10 −4 ). F) Quantification of ΔPO showing an increased difference in the preferred orientations between contralateral and ipsilateral inputs to neurons in GLT1 HET animals (n=4-6 animals, 23-52 cells per animal, t-test, p=1.0×10 −4 ). *p
    Figure Legend Snippet: GLT1 HET mice have higher ipsilateral eye responses, lower contralateral eye bias and disrupted experience-dependent binocular matching of orientation-selective responses. A) Schematic of experimental design. Top: Visual gratings were separately presented to the contra (green) and ipsi (blue) eyes in P28 mice and neuronal responses recorded. Bottom: schematic of measures. Ocular dominance index (ODI) was calculated as (max Contra − max Ipsi ) / max Contra +max Ipsi . Orientation Selectivity Index (OSI) was calculated as described previously ( Banerjee et al., 2016 ). Difference in preferred orientation (ΔPO) was calculated as the difference between preferred orientations of the max contralateral and ipsilateral responses. B) Example cells in GLT1 WT (top) and GLT1 HET (bottom) animals. Left: in vivo images of neuronal somas measured in binocular visual cortex using the calcium indicator, GCaMP6f. Right: Tuning curves of three cells (white circles in left) to contra (green) and ipsi (blue) stimulation. Note the matched tuning and contralateral bias in WT animals and the mismatched tuning curves and lack of contralateral bias in GLT1 HETs. C) Quantification of the average response to PO in GLT1 WT and HET mice. WT mice have a significantly higher contralateral response than ipsilateral response while HET mice have approximately equal contralateral and ipsilateral responses (n=4-6 animals, 23-52 cells per animal, two-way ANOVA, genotype F(1,674)=7.72, p=0.0056, interaction F(1,674)=4.243, p=0.040). D) Quantification of ocular dominance index showing that GLT1 HET mice have significantly decreased ODI (n=4-6 animals, 23-52 cells per animal, t-test, p=0.0018). E) Quantification of OSI showing that GLT1 HET mice have a significantly decreased OSI of ipsilateral responses compared to both contra and ipsi responses in GLT1 WT animals (n=4-6 animals, 23-52 cells per animal, two-way ANOVA, genotype F(1,674)=12.46, p=4.5×10 −4 ). F) Quantification of ΔPO showing an increased difference in the preferred orientations between contralateral and ipsilateral inputs to neurons in GLT1 HET animals (n=4-6 animals, 23-52 cells per animal, t-test, p=1.0×10 −4 ). *p

    Techniques Used: Mouse Assay, In Vivo

    GLT1 is upregulated in the developing visual cortex concurrent with visual experience. A) Immunohistochemical stain of GLT1 (red), astrocytes (green), and DAPI (blue) at ~P28 in mouse visual cortex. GLT1 is expressed by astrocytes throughout cortical layers. B) High-magnification image of a single GFAP-labelled astrocyte with surrounding GLT1 expression. C) Quantification of GLT1-mRNA across developmental time points showing a significant increase from birth and peaking at P21. Levels are normalized to P28 (P0 (n=4), P7 (n=4), P14 (n=3), P21 (n=3), P28 (n=4), P42 (n=3), P60 (n=4); one-way ANOVA, F(6,18)=86.7, p=2.7×10 −12 ). D) Western blot quantification showing that transgenic mice with heterozygous expression of GLT1 (HET) have significantly less GLT1 expression compared to wildtype (WT) littermates (n=WT(11), HET(11); unpaired t-test, t=4.64, p=2.7×10 −4 ), but comparable expression of GLAST (WT (n=8), HET (n=4); unpaired t-test, t=0.62, p=0.55). E) Example images of GLT1 WT and HET astrocytes labeled using a custom GFAP-tdTomato transgenic mouse line (red). Astrocyte volume is reconstruction from imaged z-stacks (green). F) Quantification of astrocyte volume from 3D reconstructions show no difference between GLT1 WT and HET animals (WT (n=3), HET (n=13); unpaired t-test, t=0.53, p=0.60). G) Images of the lateral geniculate nucleus (LGN) after CTB-594 (red) and CTB-488 (green) injection into the contralateral and ipsilateral eyes respectively. Merged overlays from GLT1 WT and HET mice show normal retinothalamic axon segregation. H) Quantification of ipsilateral area in GLT1 WT and HETs across several binary thresholds (0, 5, 30%) showing no difference in absolute ipsilateral area (WT (n=3), HET (n=6); two-way ANOVA, genotype effect F(1,7)=1.93, p=0.21). I) Quantification of contra/ipsi projection overlap showing no difference in contra/ipsi segregation (WT (n=3), HET (n=6); two-way ANOVA, genotype effect, F(1,7)=0.43, p=0.53). *p
    Figure Legend Snippet: GLT1 is upregulated in the developing visual cortex concurrent with visual experience. A) Immunohistochemical stain of GLT1 (red), astrocytes (green), and DAPI (blue) at ~P28 in mouse visual cortex. GLT1 is expressed by astrocytes throughout cortical layers. B) High-magnification image of a single GFAP-labelled astrocyte with surrounding GLT1 expression. C) Quantification of GLT1-mRNA across developmental time points showing a significant increase from birth and peaking at P21. Levels are normalized to P28 (P0 (n=4), P7 (n=4), P14 (n=3), P21 (n=3), P28 (n=4), P42 (n=3), P60 (n=4); one-way ANOVA, F(6,18)=86.7, p=2.7×10 −12 ). D) Western blot quantification showing that transgenic mice with heterozygous expression of GLT1 (HET) have significantly less GLT1 expression compared to wildtype (WT) littermates (n=WT(11), HET(11); unpaired t-test, t=4.64, p=2.7×10 −4 ), but comparable expression of GLAST (WT (n=8), HET (n=4); unpaired t-test, t=0.62, p=0.55). E) Example images of GLT1 WT and HET astrocytes labeled using a custom GFAP-tdTomato transgenic mouse line (red). Astrocyte volume is reconstruction from imaged z-stacks (green). F) Quantification of astrocyte volume from 3D reconstructions show no difference between GLT1 WT and HET animals (WT (n=3), HET (n=13); unpaired t-test, t=0.53, p=0.60). G) Images of the lateral geniculate nucleus (LGN) after CTB-594 (red) and CTB-488 (green) injection into the contralateral and ipsilateral eyes respectively. Merged overlays from GLT1 WT and HET mice show normal retinothalamic axon segregation. H) Quantification of ipsilateral area in GLT1 WT and HETs across several binary thresholds (0, 5, 30%) showing no difference in absolute ipsilateral area (WT (n=3), HET (n=6); two-way ANOVA, genotype effect F(1,7)=1.93, p=0.21). I) Quantification of contra/ipsi projection overlap showing no difference in contra/ipsi segregation (WT (n=3), HET (n=6); two-way ANOVA, genotype effect, F(1,7)=0.43, p=0.53). *p

    Techniques Used: Immunohistochemistry, Staining, Expressing, Western Blot, Transgenic Assay, Mouse Assay, Labeling, CtB Assay, Injection

    GLT1 HET mice have upregulation of GLT1 expression during monocular deprivation. A) Quantification of GLT1 mRNA in WT (grays) and HET (reds) mice in ND, 4dMD, and 7dMD conditions. GLT1 HET mice have significantly less GLT1 mRNA in ND conditions, but no difference at 4dMD and 7dMD compared to WT mice (n=2-3 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). B) Example western blots for GLT1 protein in WT (top) and HET (bottom) mice in ND, 4dMD, and 7dMD conditions. C) Quantification of western blots showing significantly less GLT1 protein in HET mice in ND, but no difference in 4dMD and 7dMD compared to WT littermates (n=4-9 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). *p
    Figure Legend Snippet: GLT1 HET mice have upregulation of GLT1 expression during monocular deprivation. A) Quantification of GLT1 mRNA in WT (grays) and HET (reds) mice in ND, 4dMD, and 7dMD conditions. GLT1 HET mice have significantly less GLT1 mRNA in ND conditions, but no difference at 4dMD and 7dMD compared to WT mice (n=2-3 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). B) Example western blots for GLT1 protein in WT (top) and HET (bottom) mice in ND, 4dMD, and 7dMD conditions. C) Quantification of western blots showing significantly less GLT1 protein in HET mice in ND, but no difference in 4dMD and 7dMD compared to WT littermates (n=4-9 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). *p

    Techniques Used: Mouse Assay, Expressing, Western Blot

    GLT1 HET mice have abnormal maturation of excitatory and inhibitory circuits. A) Left: low-magnification image of neurons in visual cortex of GFP-M transgenic mice. Right: higher-magnification image of layer 2/3 neurons (dotted box in left). B) Images of basal dendrites of layer 2/3 neurons in GLT1 WT (top) and HET (bottom) mice. WT example is from dotted box in right panel of A. C) GLT1 HET mice have increased spine density on basal dendrites of layer 2/3 neurons in visual cortex (n=4 animals, 5 slices, 10 dendrites per animal, t-test, p=0.041). D) Example traces of miniature excitatory post-synaptic currents (mEPSCs) from voltage-clamped layer 2/3 neurons in the visual cortex of GLT1 WT and HET mice. E) Quantification of mEPSC amplitude showing no difference in magnitude of mEPSCs (n=8-13 cell, t-test, p=0.49). F) Neurons from GLT1 HET mice have a trend towards increased mEPSC frequency (n=8-13 cells, t-test, p=0.052). G) Example images of parvalbumin positive (PV+, green) and somatostatin positive (SST+, magenta) interneurons in visual cortex of GLT1 WT and HET mice). H) GLT1 HET mice have a trend towards decreased PV+ neuron density (n=4 animals, 5 slices per animal, t-test, p=0.12). I) GLT1 HET mice have a significant increase in SST+ cell density (n=4 animals, 5 slices per animal, t-test, p=0.0023). J) Example images of perineuronal nets (PNNs) visualized using wisteria floribunda agglutin (WFA) staining. K) GLT1 HET mice have significantly decreased PNN density compared to WT littermates (n=9 animals, 5 slices per animal, t-test, p=0.0068). L) Model of net decrease in cortical inhibition through increased SST+ cell density inhibiting PV+ interneurons yielding increase in excitatory pyramidal neuron responses (Pyr). *p
    Figure Legend Snippet: GLT1 HET mice have abnormal maturation of excitatory and inhibitory circuits. A) Left: low-magnification image of neurons in visual cortex of GFP-M transgenic mice. Right: higher-magnification image of layer 2/3 neurons (dotted box in left). B) Images of basal dendrites of layer 2/3 neurons in GLT1 WT (top) and HET (bottom) mice. WT example is from dotted box in right panel of A. C) GLT1 HET mice have increased spine density on basal dendrites of layer 2/3 neurons in visual cortex (n=4 animals, 5 slices, 10 dendrites per animal, t-test, p=0.041). D) Example traces of miniature excitatory post-synaptic currents (mEPSCs) from voltage-clamped layer 2/3 neurons in the visual cortex of GLT1 WT and HET mice. E) Quantification of mEPSC amplitude showing no difference in magnitude of mEPSCs (n=8-13 cell, t-test, p=0.49). F) Neurons from GLT1 HET mice have a trend towards increased mEPSC frequency (n=8-13 cells, t-test, p=0.052). G) Example images of parvalbumin positive (PV+, green) and somatostatin positive (SST+, magenta) interneurons in visual cortex of GLT1 WT and HET mice). H) GLT1 HET mice have a trend towards decreased PV+ neuron density (n=4 animals, 5 slices per animal, t-test, p=0.12). I) GLT1 HET mice have a significant increase in SST+ cell density (n=4 animals, 5 slices per animal, t-test, p=0.0023). J) Example images of perineuronal nets (PNNs) visualized using wisteria floribunda agglutin (WFA) staining. K) GLT1 HET mice have significantly decreased PNN density compared to WT littermates (n=9 animals, 5 slices per animal, t-test, p=0.0068). L) Model of net decrease in cortical inhibition through increased SST+ cell density inhibiting PV+ interneurons yielding increase in excitatory pyramidal neuron responses (Pyr). *p

    Techniques Used: Mouse Assay, Transgenic Assay, Staining, Inhibition

    GLT1 HET mice have disrupted ocular dominance plasticity. A) Schematic of experimental setup for intrinsic signal optical imaging. Drifting bars are presented to each eye individually and phase maps are generated by the retinotopic activity in visual cortex. Averaged responses of multiple sweeps yield an amplitude map. Ocular Dominance Index (ODI) is calculated as the contralateral response − ipsilateral response / contralateral + ipsilateral responses. B) ODI for GLT1 WT (grays) and GLT1 HET (reds) mice that were either non-deprived (ND), or had the contralateral eye monocularly deprived for 4 days (4dMD) or 7 days (7dMD). GLT1 WT mice display a typical contralateral bias in ND conditions. After 4dMD and 7dMD the ODI significantly decreases demonstrating intact ocular dominance plasticity. GLT1 HET mice display an abnormal lack of contralateral ODI bias under ND conditions, a significant decrease in ODI at 4dMD, and a return to no bias at 7dMD (n=4 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). C) Comparison of eye-specific amplitudes for GLT1 WT and HET mice. ND GLT1 HET mice have approximately equal responses to contralateral and ipsilateral inputs. After 4dMD, GLT1 HET mice have a significant decrease in contralateral responses and at 7dMD, significant decrease in both contralateral and ipsilateral responses (n=4 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). *p
    Figure Legend Snippet: GLT1 HET mice have disrupted ocular dominance plasticity. A) Schematic of experimental setup for intrinsic signal optical imaging. Drifting bars are presented to each eye individually and phase maps are generated by the retinotopic activity in visual cortex. Averaged responses of multiple sweeps yield an amplitude map. Ocular Dominance Index (ODI) is calculated as the contralateral response − ipsilateral response / contralateral + ipsilateral responses. B) ODI for GLT1 WT (grays) and GLT1 HET (reds) mice that were either non-deprived (ND), or had the contralateral eye monocularly deprived for 4 days (4dMD) or 7 days (7dMD). GLT1 WT mice display a typical contralateral bias in ND conditions. After 4dMD and 7dMD the ODI significantly decreases demonstrating intact ocular dominance plasticity. GLT1 HET mice display an abnormal lack of contralateral ODI bias under ND conditions, a significant decrease in ODI at 4dMD, and a return to no bias at 7dMD (n=4 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). C) Comparison of eye-specific amplitudes for GLT1 WT and HET mice. ND GLT1 HET mice have approximately equal responses to contralateral and ipsilateral inputs. After 4dMD, GLT1 HET mice have a significant decrease in contralateral responses and at 7dMD, significant decrease in both contralateral and ipsilateral responses (n=4 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). *p

    Techniques Used: Mouse Assay, Optical Imaging, Generated, Activity Assay

    Summary and model of experimental results. A) Schematic showing the contralateral (contra, green) and ipsilateral (ipsi, blue) inputs to binocular visual cortex synapsing onto layer 2/3 pyramidal cells (L2/3, Pyr, gray). Astrocytes (Ast, orange) have fine processes that surround excitatory synapses. Dashed box is magnified below, with details. B) At eye-opening, GLT1 WT and HET animals have similar astrocyte volume and LGN refinement. With visual experience, GLT1-WT animals undergo activity-dependent plasticity resulting in decreased spine density, binocular matching of preferred orientation, and a contralaterally biased ocular dominance. GLT1-HET animals have comparatively decreased GLT1 protein resulting in increased dendritic spines, increased ipsi responses, reduced contra bias and orientation tuning, and decreased binocular matching of orientation preference. Following 4 days of MD in GLT1-WT mice, contralateral responses decrease and after 7 days of MD, ipsilateral responses increase. In GLT1-HET animals, after 4 days of MD contralateral responses decrease resulting in a negative ODI. However, after 7 days of MD, increased GLT1 expression also decreases ipsilateral inputs resulting in no ocular dominance bias. These results are reasonably explained by a selective influence of GLT1 on ipsilateral inputs and responses during development.
    Figure Legend Snippet: Summary and model of experimental results. A) Schematic showing the contralateral (contra, green) and ipsilateral (ipsi, blue) inputs to binocular visual cortex synapsing onto layer 2/3 pyramidal cells (L2/3, Pyr, gray). Astrocytes (Ast, orange) have fine processes that surround excitatory synapses. Dashed box is magnified below, with details. B) At eye-opening, GLT1 WT and HET animals have similar astrocyte volume and LGN refinement. With visual experience, GLT1-WT animals undergo activity-dependent plasticity resulting in decreased spine density, binocular matching of preferred orientation, and a contralaterally biased ocular dominance. GLT1-HET animals have comparatively decreased GLT1 protein resulting in increased dendritic spines, increased ipsi responses, reduced contra bias and orientation tuning, and decreased binocular matching of orientation preference. Following 4 days of MD in GLT1-WT mice, contralateral responses decrease and after 7 days of MD, ipsilateral responses increase. In GLT1-HET animals, after 4 days of MD contralateral responses decrease resulting in a negative ODI. However, after 7 days of MD, increased GLT1 expression also decreases ipsilateral inputs resulting in no ocular dominance bias. These results are reasonably explained by a selective influence of GLT1 on ipsilateral inputs and responses during development.

    Techniques Used: AST Assay, Activity Assay, Mouse Assay, Expressing

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    Alomone Labs rabbit α glt1
    <t>GLT1</t> HET mice have higher ipsilateral eye responses, lower contralateral eye bias and disrupted experience-dependent binocular matching of orientation-selective responses. A) Schematic of experimental design. Top: Visual gratings were separately presented to the contra (green) and ipsi (blue) eyes in P28 mice and neuronal responses recorded. Bottom: schematic of measures. Ocular dominance index (ODI) was calculated as (max Contra − max Ipsi ) / max Contra +max Ipsi . Orientation Selectivity Index (OSI) was calculated as described previously ( Banerjee et al., 2016 ). Difference in preferred orientation (ΔPO) was calculated as the difference between preferred orientations of the max contralateral and ipsilateral responses. B) Example cells in GLT1 WT (top) and GLT1 HET (bottom) animals. Left: in vivo images of neuronal somas measured in binocular visual cortex using the calcium indicator, GCaMP6f. Right: Tuning curves of three cells (white circles in left) to contra (green) and ipsi (blue) stimulation. Note the matched tuning and contralateral bias in WT animals and the mismatched tuning curves and lack of contralateral bias in GLT1 HETs. C) Quantification of the average response to PO in GLT1 WT and HET mice. WT mice have a significantly higher contralateral response than ipsilateral response while HET mice have approximately equal contralateral and ipsilateral responses (n=4-6 animals, 23-52 cells per animal, two-way ANOVA, genotype F(1,674)=7.72, p=0.0056, interaction F(1,674)=4.243, p=0.040). D) Quantification of ocular dominance index showing that GLT1 HET mice have significantly decreased ODI (n=4-6 animals, 23-52 cells per animal, t-test, p=0.0018). E) Quantification of OSI showing that GLT1 HET mice have a significantly decreased OSI of ipsilateral responses compared to both contra and ipsi responses in GLT1 WT animals (n=4-6 animals, 23-52 cells per animal, two-way ANOVA, genotype F(1,674)=12.46, p=4.5×10 −4 ). F) Quantification of ΔPO showing an increased difference in the preferred orientations between contralateral and ipsilateral inputs to neurons in GLT1 HET animals (n=4-6 animals, 23-52 cells per animal, t-test, p=1.0×10 −4 ). *p
    Rabbit α Glt1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit α glt1/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit α glt1 - by Bioz Stars, 2022-12
    94/100 stars
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    GLT1 HET mice have higher ipsilateral eye responses, lower contralateral eye bias and disrupted experience-dependent binocular matching of orientation-selective responses. A) Schematic of experimental design. Top: Visual gratings were separately presented to the contra (green) and ipsi (blue) eyes in P28 mice and neuronal responses recorded. Bottom: schematic of measures. Ocular dominance index (ODI) was calculated as (max Contra − max Ipsi ) / max Contra +max Ipsi . Orientation Selectivity Index (OSI) was calculated as described previously ( Banerjee et al., 2016 ). Difference in preferred orientation (ΔPO) was calculated as the difference between preferred orientations of the max contralateral and ipsilateral responses. B) Example cells in GLT1 WT (top) and GLT1 HET (bottom) animals. Left: in vivo images of neuronal somas measured in binocular visual cortex using the calcium indicator, GCaMP6f. Right: Tuning curves of three cells (white circles in left) to contra (green) and ipsi (blue) stimulation. Note the matched tuning and contralateral bias in WT animals and the mismatched tuning curves and lack of contralateral bias in GLT1 HETs. C) Quantification of the average response to PO in GLT1 WT and HET mice. WT mice have a significantly higher contralateral response than ipsilateral response while HET mice have approximately equal contralateral and ipsilateral responses (n=4-6 animals, 23-52 cells per animal, two-way ANOVA, genotype F(1,674)=7.72, p=0.0056, interaction F(1,674)=4.243, p=0.040). D) Quantification of ocular dominance index showing that GLT1 HET mice have significantly decreased ODI (n=4-6 animals, 23-52 cells per animal, t-test, p=0.0018). E) Quantification of OSI showing that GLT1 HET mice have a significantly decreased OSI of ipsilateral responses compared to both contra and ipsi responses in GLT1 WT animals (n=4-6 animals, 23-52 cells per animal, two-way ANOVA, genotype F(1,674)=12.46, p=4.5×10 −4 ). F) Quantification of ΔPO showing an increased difference in the preferred orientations between contralateral and ipsilateral inputs to neurons in GLT1 HET animals (n=4-6 animals, 23-52 cells per animal, t-test, p=1.0×10 −4 ). *p

    Journal: bioRxiv

    Article Title: Astrocytic glutamate uptake coordinates experience-dependent, eye-specific refinement in developing visual cortex

    doi: 10.1101/2020.05.25.113613

    Figure Lengend Snippet: GLT1 HET mice have higher ipsilateral eye responses, lower contralateral eye bias and disrupted experience-dependent binocular matching of orientation-selective responses. A) Schematic of experimental design. Top: Visual gratings were separately presented to the contra (green) and ipsi (blue) eyes in P28 mice and neuronal responses recorded. Bottom: schematic of measures. Ocular dominance index (ODI) was calculated as (max Contra − max Ipsi ) / max Contra +max Ipsi . Orientation Selectivity Index (OSI) was calculated as described previously ( Banerjee et al., 2016 ). Difference in preferred orientation (ΔPO) was calculated as the difference between preferred orientations of the max contralateral and ipsilateral responses. B) Example cells in GLT1 WT (top) and GLT1 HET (bottom) animals. Left: in vivo images of neuronal somas measured in binocular visual cortex using the calcium indicator, GCaMP6f. Right: Tuning curves of three cells (white circles in left) to contra (green) and ipsi (blue) stimulation. Note the matched tuning and contralateral bias in WT animals and the mismatched tuning curves and lack of contralateral bias in GLT1 HETs. C) Quantification of the average response to PO in GLT1 WT and HET mice. WT mice have a significantly higher contralateral response than ipsilateral response while HET mice have approximately equal contralateral and ipsilateral responses (n=4-6 animals, 23-52 cells per animal, two-way ANOVA, genotype F(1,674)=7.72, p=0.0056, interaction F(1,674)=4.243, p=0.040). D) Quantification of ocular dominance index showing that GLT1 HET mice have significantly decreased ODI (n=4-6 animals, 23-52 cells per animal, t-test, p=0.0018). E) Quantification of OSI showing that GLT1 HET mice have a significantly decreased OSI of ipsilateral responses compared to both contra and ipsi responses in GLT1 WT animals (n=4-6 animals, 23-52 cells per animal, two-way ANOVA, genotype F(1,674)=12.46, p=4.5×10 −4 ). F) Quantification of ΔPO showing an increased difference in the preferred orientations between contralateral and ipsilateral inputs to neurons in GLT1 HET animals (n=4-6 animals, 23-52 cells per animal, t-test, p=1.0×10 −4 ). *p

    Article Snippet: The following primary antibodies and dilutions were used: rabbit α-GLT1 (1:500, AGC-022, Alamone Labs), guinea pig α-GFAP (1:2000, 173 004, Synaptic Systems), chicken α-GFP (1:1000, GFP-1020, Aves Labs), α-rabbit α-SST-14 (1:500, T-4103, Peninsula Labs), mouse α-PV (1:500, MAB1572, EMD Millipore).

    Techniques: Mouse Assay, In Vivo

    GLT1 is upregulated in the developing visual cortex concurrent with visual experience. A) Immunohistochemical stain of GLT1 (red), astrocytes (green), and DAPI (blue) at ~P28 in mouse visual cortex. GLT1 is expressed by astrocytes throughout cortical layers. B) High-magnification image of a single GFAP-labelled astrocyte with surrounding GLT1 expression. C) Quantification of GLT1-mRNA across developmental time points showing a significant increase from birth and peaking at P21. Levels are normalized to P28 (P0 (n=4), P7 (n=4), P14 (n=3), P21 (n=3), P28 (n=4), P42 (n=3), P60 (n=4); one-way ANOVA, F(6,18)=86.7, p=2.7×10 −12 ). D) Western blot quantification showing that transgenic mice with heterozygous expression of GLT1 (HET) have significantly less GLT1 expression compared to wildtype (WT) littermates (n=WT(11), HET(11); unpaired t-test, t=4.64, p=2.7×10 −4 ), but comparable expression of GLAST (WT (n=8), HET (n=4); unpaired t-test, t=0.62, p=0.55). E) Example images of GLT1 WT and HET astrocytes labeled using a custom GFAP-tdTomato transgenic mouse line (red). Astrocyte volume is reconstruction from imaged z-stacks (green). F) Quantification of astrocyte volume from 3D reconstructions show no difference between GLT1 WT and HET animals (WT (n=3), HET (n=13); unpaired t-test, t=0.53, p=0.60). G) Images of the lateral geniculate nucleus (LGN) after CTB-594 (red) and CTB-488 (green) injection into the contralateral and ipsilateral eyes respectively. Merged overlays from GLT1 WT and HET mice show normal retinothalamic axon segregation. H) Quantification of ipsilateral area in GLT1 WT and HETs across several binary thresholds (0, 5, 30%) showing no difference in absolute ipsilateral area (WT (n=3), HET (n=6); two-way ANOVA, genotype effect F(1,7)=1.93, p=0.21). I) Quantification of contra/ipsi projection overlap showing no difference in contra/ipsi segregation (WT (n=3), HET (n=6); two-way ANOVA, genotype effect, F(1,7)=0.43, p=0.53). *p

    Journal: bioRxiv

    Article Title: Astrocytic glutamate uptake coordinates experience-dependent, eye-specific refinement in developing visual cortex

    doi: 10.1101/2020.05.25.113613

    Figure Lengend Snippet: GLT1 is upregulated in the developing visual cortex concurrent with visual experience. A) Immunohistochemical stain of GLT1 (red), astrocytes (green), and DAPI (blue) at ~P28 in mouse visual cortex. GLT1 is expressed by astrocytes throughout cortical layers. B) High-magnification image of a single GFAP-labelled astrocyte with surrounding GLT1 expression. C) Quantification of GLT1-mRNA across developmental time points showing a significant increase from birth and peaking at P21. Levels are normalized to P28 (P0 (n=4), P7 (n=4), P14 (n=3), P21 (n=3), P28 (n=4), P42 (n=3), P60 (n=4); one-way ANOVA, F(6,18)=86.7, p=2.7×10 −12 ). D) Western blot quantification showing that transgenic mice with heterozygous expression of GLT1 (HET) have significantly less GLT1 expression compared to wildtype (WT) littermates (n=WT(11), HET(11); unpaired t-test, t=4.64, p=2.7×10 −4 ), but comparable expression of GLAST (WT (n=8), HET (n=4); unpaired t-test, t=0.62, p=0.55). E) Example images of GLT1 WT and HET astrocytes labeled using a custom GFAP-tdTomato transgenic mouse line (red). Astrocyte volume is reconstruction from imaged z-stacks (green). F) Quantification of astrocyte volume from 3D reconstructions show no difference between GLT1 WT and HET animals (WT (n=3), HET (n=13); unpaired t-test, t=0.53, p=0.60). G) Images of the lateral geniculate nucleus (LGN) after CTB-594 (red) and CTB-488 (green) injection into the contralateral and ipsilateral eyes respectively. Merged overlays from GLT1 WT and HET mice show normal retinothalamic axon segregation. H) Quantification of ipsilateral area in GLT1 WT and HETs across several binary thresholds (0, 5, 30%) showing no difference in absolute ipsilateral area (WT (n=3), HET (n=6); two-way ANOVA, genotype effect F(1,7)=1.93, p=0.21). I) Quantification of contra/ipsi projection overlap showing no difference in contra/ipsi segregation (WT (n=3), HET (n=6); two-way ANOVA, genotype effect, F(1,7)=0.43, p=0.53). *p

    Article Snippet: The following primary antibodies and dilutions were used: rabbit α-GLT1 (1:500, AGC-022, Alamone Labs), guinea pig α-GFAP (1:2000, 173 004, Synaptic Systems), chicken α-GFP (1:1000, GFP-1020, Aves Labs), α-rabbit α-SST-14 (1:500, T-4103, Peninsula Labs), mouse α-PV (1:500, MAB1572, EMD Millipore).

    Techniques: Immunohistochemistry, Staining, Expressing, Western Blot, Transgenic Assay, Mouse Assay, Labeling, CtB Assay, Injection

    GLT1 HET mice have upregulation of GLT1 expression during monocular deprivation. A) Quantification of GLT1 mRNA in WT (grays) and HET (reds) mice in ND, 4dMD, and 7dMD conditions. GLT1 HET mice have significantly less GLT1 mRNA in ND conditions, but no difference at 4dMD and 7dMD compared to WT mice (n=2-3 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). B) Example western blots for GLT1 protein in WT (top) and HET (bottom) mice in ND, 4dMD, and 7dMD conditions. C) Quantification of western blots showing significantly less GLT1 protein in HET mice in ND, but no difference in 4dMD and 7dMD compared to WT littermates (n=4-9 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). *p

    Journal: bioRxiv

    Article Title: Astrocytic glutamate uptake coordinates experience-dependent, eye-specific refinement in developing visual cortex

    doi: 10.1101/2020.05.25.113613

    Figure Lengend Snippet: GLT1 HET mice have upregulation of GLT1 expression during monocular deprivation. A) Quantification of GLT1 mRNA in WT (grays) and HET (reds) mice in ND, 4dMD, and 7dMD conditions. GLT1 HET mice have significantly less GLT1 mRNA in ND conditions, but no difference at 4dMD and 7dMD compared to WT mice (n=2-3 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). B) Example western blots for GLT1 protein in WT (top) and HET (bottom) mice in ND, 4dMD, and 7dMD conditions. C) Quantification of western blots showing significantly less GLT1 protein in HET mice in ND, but no difference in 4dMD and 7dMD compared to WT littermates (n=4-9 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). *p

    Article Snippet: The following primary antibodies and dilutions were used: rabbit α-GLT1 (1:500, AGC-022, Alamone Labs), guinea pig α-GFAP (1:2000, 173 004, Synaptic Systems), chicken α-GFP (1:1000, GFP-1020, Aves Labs), α-rabbit α-SST-14 (1:500, T-4103, Peninsula Labs), mouse α-PV (1:500, MAB1572, EMD Millipore).

    Techniques: Mouse Assay, Expressing, Western Blot

    GLT1 HET mice have abnormal maturation of excitatory and inhibitory circuits. A) Left: low-magnification image of neurons in visual cortex of GFP-M transgenic mice. Right: higher-magnification image of layer 2/3 neurons (dotted box in left). B) Images of basal dendrites of layer 2/3 neurons in GLT1 WT (top) and HET (bottom) mice. WT example is from dotted box in right panel of A. C) GLT1 HET mice have increased spine density on basal dendrites of layer 2/3 neurons in visual cortex (n=4 animals, 5 slices, 10 dendrites per animal, t-test, p=0.041). D) Example traces of miniature excitatory post-synaptic currents (mEPSCs) from voltage-clamped layer 2/3 neurons in the visual cortex of GLT1 WT and HET mice. E) Quantification of mEPSC amplitude showing no difference in magnitude of mEPSCs (n=8-13 cell, t-test, p=0.49). F) Neurons from GLT1 HET mice have a trend towards increased mEPSC frequency (n=8-13 cells, t-test, p=0.052). G) Example images of parvalbumin positive (PV+, green) and somatostatin positive (SST+, magenta) interneurons in visual cortex of GLT1 WT and HET mice). H) GLT1 HET mice have a trend towards decreased PV+ neuron density (n=4 animals, 5 slices per animal, t-test, p=0.12). I) GLT1 HET mice have a significant increase in SST+ cell density (n=4 animals, 5 slices per animal, t-test, p=0.0023). J) Example images of perineuronal nets (PNNs) visualized using wisteria floribunda agglutin (WFA) staining. K) GLT1 HET mice have significantly decreased PNN density compared to WT littermates (n=9 animals, 5 slices per animal, t-test, p=0.0068). L) Model of net decrease in cortical inhibition through increased SST+ cell density inhibiting PV+ interneurons yielding increase in excitatory pyramidal neuron responses (Pyr). *p

    Journal: bioRxiv

    Article Title: Astrocytic glutamate uptake coordinates experience-dependent, eye-specific refinement in developing visual cortex

    doi: 10.1101/2020.05.25.113613

    Figure Lengend Snippet: GLT1 HET mice have abnormal maturation of excitatory and inhibitory circuits. A) Left: low-magnification image of neurons in visual cortex of GFP-M transgenic mice. Right: higher-magnification image of layer 2/3 neurons (dotted box in left). B) Images of basal dendrites of layer 2/3 neurons in GLT1 WT (top) and HET (bottom) mice. WT example is from dotted box in right panel of A. C) GLT1 HET mice have increased spine density on basal dendrites of layer 2/3 neurons in visual cortex (n=4 animals, 5 slices, 10 dendrites per animal, t-test, p=0.041). D) Example traces of miniature excitatory post-synaptic currents (mEPSCs) from voltage-clamped layer 2/3 neurons in the visual cortex of GLT1 WT and HET mice. E) Quantification of mEPSC amplitude showing no difference in magnitude of mEPSCs (n=8-13 cell, t-test, p=0.49). F) Neurons from GLT1 HET mice have a trend towards increased mEPSC frequency (n=8-13 cells, t-test, p=0.052). G) Example images of parvalbumin positive (PV+, green) and somatostatin positive (SST+, magenta) interneurons in visual cortex of GLT1 WT and HET mice). H) GLT1 HET mice have a trend towards decreased PV+ neuron density (n=4 animals, 5 slices per animal, t-test, p=0.12). I) GLT1 HET mice have a significant increase in SST+ cell density (n=4 animals, 5 slices per animal, t-test, p=0.0023). J) Example images of perineuronal nets (PNNs) visualized using wisteria floribunda agglutin (WFA) staining. K) GLT1 HET mice have significantly decreased PNN density compared to WT littermates (n=9 animals, 5 slices per animal, t-test, p=0.0068). L) Model of net decrease in cortical inhibition through increased SST+ cell density inhibiting PV+ interneurons yielding increase in excitatory pyramidal neuron responses (Pyr). *p

    Article Snippet: The following primary antibodies and dilutions were used: rabbit α-GLT1 (1:500, AGC-022, Alamone Labs), guinea pig α-GFAP (1:2000, 173 004, Synaptic Systems), chicken α-GFP (1:1000, GFP-1020, Aves Labs), α-rabbit α-SST-14 (1:500, T-4103, Peninsula Labs), mouse α-PV (1:500, MAB1572, EMD Millipore).

    Techniques: Mouse Assay, Transgenic Assay, Staining, Inhibition

    GLT1 HET mice have disrupted ocular dominance plasticity. A) Schematic of experimental setup for intrinsic signal optical imaging. Drifting bars are presented to each eye individually and phase maps are generated by the retinotopic activity in visual cortex. Averaged responses of multiple sweeps yield an amplitude map. Ocular Dominance Index (ODI) is calculated as the contralateral response − ipsilateral response / contralateral + ipsilateral responses. B) ODI for GLT1 WT (grays) and GLT1 HET (reds) mice that were either non-deprived (ND), or had the contralateral eye monocularly deprived for 4 days (4dMD) or 7 days (7dMD). GLT1 WT mice display a typical contralateral bias in ND conditions. After 4dMD and 7dMD the ODI significantly decreases demonstrating intact ocular dominance plasticity. GLT1 HET mice display an abnormal lack of contralateral ODI bias under ND conditions, a significant decrease in ODI at 4dMD, and a return to no bias at 7dMD (n=4 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). C) Comparison of eye-specific amplitudes for GLT1 WT and HET mice. ND GLT1 HET mice have approximately equal responses to contralateral and ipsilateral inputs. After 4dMD, GLT1 HET mice have a significant decrease in contralateral responses and at 7dMD, significant decrease in both contralateral and ipsilateral responses (n=4 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). *p

    Journal: bioRxiv

    Article Title: Astrocytic glutamate uptake coordinates experience-dependent, eye-specific refinement in developing visual cortex

    doi: 10.1101/2020.05.25.113613

    Figure Lengend Snippet: GLT1 HET mice have disrupted ocular dominance plasticity. A) Schematic of experimental setup for intrinsic signal optical imaging. Drifting bars are presented to each eye individually and phase maps are generated by the retinotopic activity in visual cortex. Averaged responses of multiple sweeps yield an amplitude map. Ocular Dominance Index (ODI) is calculated as the contralateral response − ipsilateral response / contralateral + ipsilateral responses. B) ODI for GLT1 WT (grays) and GLT1 HET (reds) mice that were either non-deprived (ND), or had the contralateral eye monocularly deprived for 4 days (4dMD) or 7 days (7dMD). GLT1 WT mice display a typical contralateral bias in ND conditions. After 4dMD and 7dMD the ODI significantly decreases demonstrating intact ocular dominance plasticity. GLT1 HET mice display an abnormal lack of contralateral ODI bias under ND conditions, a significant decrease in ODI at 4dMD, and a return to no bias at 7dMD (n=4 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). C) Comparison of eye-specific amplitudes for GLT1 WT and HET mice. ND GLT1 HET mice have approximately equal responses to contralateral and ipsilateral inputs. After 4dMD, GLT1 HET mice have a significant decrease in contralateral responses and at 7dMD, significant decrease in both contralateral and ipsilateral responses (n=4 animals per group, two-way ANOVA, Holm-Sidak post-hoc comparisons). *p

    Article Snippet: The following primary antibodies and dilutions were used: rabbit α-GLT1 (1:500, AGC-022, Alamone Labs), guinea pig α-GFAP (1:2000, 173 004, Synaptic Systems), chicken α-GFP (1:1000, GFP-1020, Aves Labs), α-rabbit α-SST-14 (1:500, T-4103, Peninsula Labs), mouse α-PV (1:500, MAB1572, EMD Millipore).

    Techniques: Mouse Assay, Optical Imaging, Generated, Activity Assay

    Summary and model of experimental results. A) Schematic showing the contralateral (contra, green) and ipsilateral (ipsi, blue) inputs to binocular visual cortex synapsing onto layer 2/3 pyramidal cells (L2/3, Pyr, gray). Astrocytes (Ast, orange) have fine processes that surround excitatory synapses. Dashed box is magnified below, with details. B) At eye-opening, GLT1 WT and HET animals have similar astrocyte volume and LGN refinement. With visual experience, GLT1-WT animals undergo activity-dependent plasticity resulting in decreased spine density, binocular matching of preferred orientation, and a contralaterally biased ocular dominance. GLT1-HET animals have comparatively decreased GLT1 protein resulting in increased dendritic spines, increased ipsi responses, reduced contra bias and orientation tuning, and decreased binocular matching of orientation preference. Following 4 days of MD in GLT1-WT mice, contralateral responses decrease and after 7 days of MD, ipsilateral responses increase. In GLT1-HET animals, after 4 days of MD contralateral responses decrease resulting in a negative ODI. However, after 7 days of MD, increased GLT1 expression also decreases ipsilateral inputs resulting in no ocular dominance bias. These results are reasonably explained by a selective influence of GLT1 on ipsilateral inputs and responses during development.

    Journal: bioRxiv

    Article Title: Astrocytic glutamate uptake coordinates experience-dependent, eye-specific refinement in developing visual cortex

    doi: 10.1101/2020.05.25.113613

    Figure Lengend Snippet: Summary and model of experimental results. A) Schematic showing the contralateral (contra, green) and ipsilateral (ipsi, blue) inputs to binocular visual cortex synapsing onto layer 2/3 pyramidal cells (L2/3, Pyr, gray). Astrocytes (Ast, orange) have fine processes that surround excitatory synapses. Dashed box is magnified below, with details. B) At eye-opening, GLT1 WT and HET animals have similar astrocyte volume and LGN refinement. With visual experience, GLT1-WT animals undergo activity-dependent plasticity resulting in decreased spine density, binocular matching of preferred orientation, and a contralaterally biased ocular dominance. GLT1-HET animals have comparatively decreased GLT1 protein resulting in increased dendritic spines, increased ipsi responses, reduced contra bias and orientation tuning, and decreased binocular matching of orientation preference. Following 4 days of MD in GLT1-WT mice, contralateral responses decrease and after 7 days of MD, ipsilateral responses increase. In GLT1-HET animals, after 4 days of MD contralateral responses decrease resulting in a negative ODI. However, after 7 days of MD, increased GLT1 expression also decreases ipsilateral inputs resulting in no ocular dominance bias. These results are reasonably explained by a selective influence of GLT1 on ipsilateral inputs and responses during development.

    Article Snippet: The following primary antibodies and dilutions were used: rabbit α-GLT1 (1:500, AGC-022, Alamone Labs), guinea pig α-GFAP (1:2000, 173 004, Synaptic Systems), chicken α-GFP (1:1000, GFP-1020, Aves Labs), α-rabbit α-SST-14 (1:500, T-4103, Peninsula Labs), mouse α-PV (1:500, MAB1572, EMD Millipore).

    Techniques: AST Assay, Activity Assay, Mouse Assay, Expressing