guinea pig polyclonal anti-gad2 Search Results


rabbit anti gad2  (Proteintech)
93
Proteintech rabbit anti gad2
Rabbit Anti Gad2, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mouse anti-bassoon  (BioLynx Inc)
90
BioLynx Inc mouse anti-bassoon
Mouse Anti Bassoon, supplied by BioLynx Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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sheep anti gfp  (Bio-Rad)
95
Bio-Rad sheep anti gfp
Sheep Anti Gfp, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti glutamate decarboxylase 65 gad2  (Cell Signaling Technology Inc)
93
Cell Signaling Technology Inc anti glutamate decarboxylase 65 gad2
Cortical deletion of SERT results in synaptic hyperinnervation of the DRN. a Diagram summarizing the excitatory glutamate and inhibitory GABAergic synaptic inputs received by the dorsal raphe nucleus (DRN) neurons. Synaptic inputs can be selectively identified in array tomography by the presence of specific synaptic markers including the vesicular glutamate transporter type 1 and 2 (VGLUT1 and VGLUT2, respectively) and the enzyme responsible for GABA synthesis, the <t>glutamate</t> <t>decarboxylase</t> <t>2</t> <t>(GAD2).</t> The prefrontal cortex (PFC), lateral habenula (LHb), laterodorsal tegmental nucleus (LDTg), ventral tegmental area (VTA), substantia nigra (SN), rostromedial tegmental nucleus (RMTg), periaqueductal gray (PAG), and hypothalamus (Hyp), have been noted as the main synaptic inputs to the DRN [ , , ]. b Immunolabeling against the 5-HT biosynthetic enzyme tryptophan hydroxylase (TPH) illustrating the distribution of 5-HT neurons in the midbrain DRN. The bracketed area shows the sampling region in the midline DRN used for array tomography quantitative analyses (at P28). c Array tomography projection image of three serial-ultrathin 70-nm-thick sections of the DRN immunolabeled against VGLUT1 (green) and synapsin (red) to specifically identify cortical synaptic boutons. The arrows indicate double-labeled boutons in control and SERT-KO mice. d-e Quantitative analysis of cortical glutamate synaptic boutons (VGLUT1+) ( d ), and subcortical glutamate (VGLUT2+) and GABAergic (GAD2+) synaptic boutons ( e ) in the DRN of control and SERT-KO mice (4 mice/genotype; for VGLUT1/Synapsin pairs: F 1,6 = 36.45, * p < 0.001; for VGLUT2/Synapsin pairs: F 1,6 = 0.24, p = 0.64; and for GAD2/Synapsin pairs: F 1,6 = 0.34, p = 0.58). f Analysis of cortical synaptic boutons in the DRN after fluoxetine-treatment (FLX) during the postnatal critical period (P2-14) (5 mice/group; F 1,8 = 9.94, * p < 0.02). g-h Density of cortical synaptic boutons in the DRN after conditional SERT invalidation using Emx1b Cre/+ :Sert fl/fl mice (SERT-KO CTX ) ( g ) and Pet1 Cre :Sert fl/fl mice (SERT-KO Raphe ) ( h ). ( g): 5 mice/genotype (Welch’s statistic = 11.20, * p < 0.03); ( h ): 3–4 mice/genotype (F 1,5 = 3.76, p = 0.11). i Cortical synaptic boutons in the mediodorsal thalamic nucleus (MD) of control and SERT-KO mice (3 mice/genotype; F 1,4 = 18.16, * p < 0.02). One-way ANOVA ( d, e, f, h, i ), and Welch’s t test ( g ). Error bars represent SEM
Anti Glutamate Decarboxylase 65 Gad2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/guinea+pig+polyclonal+anti-gad2/GAD2+XP+Rabbit+mAb/pmc06445781-87-28-34
Average 93 stars, based on 1 article reviews
anti glutamate decarboxylase 65 gad2 - by Bioz Stars, 2026-09
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rabbit anti-gfap  (EnCor Biotechnology)
90
EnCor Biotechnology rabbit anti-gfap
Cortical deletion of SERT results in synaptic hyperinnervation of the DRN. a Diagram summarizing the excitatory glutamate and inhibitory GABAergic synaptic inputs received by the dorsal raphe nucleus (DRN) neurons. Synaptic inputs can be selectively identified in array tomography by the presence of specific synaptic markers including the vesicular glutamate transporter type 1 and 2 (VGLUT1 and VGLUT2, respectively) and the enzyme responsible for GABA synthesis, the <t>glutamate</t> <t>decarboxylase</t> <t>2</t> <t>(GAD2).</t> The prefrontal cortex (PFC), lateral habenula (LHb), laterodorsal tegmental nucleus (LDTg), ventral tegmental area (VTA), substantia nigra (SN), rostromedial tegmental nucleus (RMTg), periaqueductal gray (PAG), and hypothalamus (Hyp), have been noted as the main synaptic inputs to the DRN [ , , ]. b Immunolabeling against the 5-HT biosynthetic enzyme tryptophan hydroxylase (TPH) illustrating the distribution of 5-HT neurons in the midbrain DRN. The bracketed area shows the sampling region in the midline DRN used for array tomography quantitative analyses (at P28). c Array tomography projection image of three serial-ultrathin 70-nm-thick sections of the DRN immunolabeled against VGLUT1 (green) and synapsin (red) to specifically identify cortical synaptic boutons. The arrows indicate double-labeled boutons in control and SERT-KO mice. d-e Quantitative analysis of cortical glutamate synaptic boutons (VGLUT1+) ( d ), and subcortical glutamate (VGLUT2+) and GABAergic (GAD2+) synaptic boutons ( e ) in the DRN of control and SERT-KO mice (4 mice/genotype; for VGLUT1/Synapsin pairs: F 1,6 = 36.45, * p < 0.001; for VGLUT2/Synapsin pairs: F 1,6 = 0.24, p = 0.64; and for GAD2/Synapsin pairs: F 1,6 = 0.34, p = 0.58). f Analysis of cortical synaptic boutons in the DRN after fluoxetine-treatment (FLX) during the postnatal critical period (P2-14) (5 mice/group; F 1,8 = 9.94, * p < 0.02). g-h Density of cortical synaptic boutons in the DRN after conditional SERT invalidation using Emx1b Cre/+ :Sert fl/fl mice (SERT-KO CTX ) ( g ) and Pet1 Cre :Sert fl/fl mice (SERT-KO Raphe ) ( h ). ( g): 5 mice/genotype (Welch’s statistic = 11.20, * p < 0.03); ( h ): 3–4 mice/genotype (F 1,5 = 3.76, p = 0.11). i Cortical synaptic boutons in the mediodorsal thalamic nucleus (MD) of control and SERT-KO mice (3 mice/genotype; F 1,4 = 18.16, * p < 0.02). One-way ANOVA ( d, e, f, h, i ), and Welch’s t test ( g ). Error bars represent SEM
Rabbit Anti Gfap, supplied by EnCor Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 90 stars, based on 1 article reviews
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mouse anti-βii spectrin antibody  (Becton Dickinson)
90
Becton Dickinson mouse anti-βii spectrin antibody
Immunolabeling of excitatory and inhibitory neurons with specific markers. (A-1 and A-2) Hippocampal neurons were fixed at DIV 10 and immunostained against vGlut1 (A-1: an excitatory neuron maker) as well as MAP2 (A-2: green) and <t>βII</t> <t>spectrin</t> (A-2: red). The specificity of vGlut1 staining is evident from the fluorescent intensity contrast observed between the arrow-marked neurons and other neurons in the field of view (A-1). (B-1 and B-2) Similar to A-1 and A-2 except that the neurons were immunostained against parvalbumin, an inhibitory neuron marker. (C-1 and C-2) Similar to A-1 and A-2 except the neurons were immunostained against GAD2, an inhibitory neuron marker. (Scale bar: 50 µm.)
Mouse Anti βii Spectrin Antibody, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/guinea+pig+polyclonal+anti-gad2/mouse+anti+%CE%B2ii+spectrin/pmc04889411-358-16-20
Average 90 stars, based on 1 article reviews
mouse anti-βii spectrin antibody - by Bioz Stars, 2026-09
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rabbit anti-rfp  (MBL International)
90
MBL International rabbit anti-rfp
Immunolabeling of excitatory and inhibitory neurons with specific markers. (A-1 and A-2) Hippocampal neurons were fixed at DIV 10 and immunostained against vGlut1 (A-1: an excitatory neuron maker) as well as MAP2 (A-2: green) and <t>βII</t> <t>spectrin</t> (A-2: red). The specificity of vGlut1 staining is evident from the fluorescent intensity contrast observed between the arrow-marked neurons and other neurons in the field of view (A-1). (B-1 and B-2) Similar to A-1 and A-2 except that the neurons were immunostained against parvalbumin, an inhibitory neuron marker. (C-1 and C-2) Similar to A-1 and A-2 except the neurons were immunostained against GAD2, an inhibitory neuron marker. (Scale bar: 50 µm.)
Rabbit Anti Rfp, supplied by MBL International, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/guinea+pig+polyclonal+anti-gad2/rabbit+anti+rfp/bio_rxiv__2022__07__04__498712-114-92-96
Average 90 stars, based on 1 article reviews
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rabbit anti omp antibody  (Santa Cruz Biotechnology)
94
Santa Cruz Biotechnology rabbit anti omp antibody
Immunolabeling of excitatory and inhibitory neurons with specific markers. (A-1 and A-2) Hippocampal neurons were fixed at DIV 10 and immunostained against vGlut1 (A-1: an excitatory neuron maker) as well as MAP2 (A-2: green) and <t>βII</t> <t>spectrin</t> (A-2: red). The specificity of vGlut1 staining is evident from the fluorescent intensity contrast observed between the arrow-marked neurons and other neurons in the field of view (A-1). (B-1 and B-2) Similar to A-1 and A-2 except that the neurons were immunostained against parvalbumin, an inhibitory neuron marker. (C-1 and C-2) Similar to A-1 and A-2 except the neurons were immunostained against GAD2, an inhibitory neuron marker. (Scale bar: 50 µm.)
Rabbit Anti Omp Antibody, supplied by Santa Cruz Biotechnology, 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/product/guinea+pig+polyclonal+anti-gad2/OMP+Antibody/pmc04889411-280-80-83
Average 94 stars, based on 1 article reviews
rabbit anti omp antibody - by Bioz Stars, 2026-09
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goat anti rfp  (Biorbyt)
93
Biorbyt goat anti rfp
Immunolabeling of excitatory and inhibitory neurons with specific markers. (A-1 and A-2) Hippocampal neurons were fixed at DIV 10 and immunostained against vGlut1 (A-1: an excitatory neuron maker) as well as MAP2 (A-2: green) and <t>βII</t> <t>spectrin</t> (A-2: red). The specificity of vGlut1 staining is evident from the fluorescent intensity contrast observed between the arrow-marked neurons and other neurons in the field of view (A-1). (B-1 and B-2) Similar to A-1 and A-2 except that the neurons were immunostained against parvalbumin, an inhibitory neuron marker. (C-1 and C-2) Similar to A-1 and A-2 except the neurons were immunostained against GAD2, an inhibitory neuron marker. (Scale bar: 50 µm.)
Goat Anti Rfp, supplied by Biorbyt, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/guinea+pig+polyclonal+anti-gad2/RFP+antibody/bio_rxiv__2022__07__04__498712-114-85-89
Average 93 stars, based on 1 article reviews
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rabbit anti eyfp  (Biorbyt)
93
Biorbyt rabbit anti eyfp
Immunolabeling of excitatory and inhibitory neurons with specific markers. (A-1 and A-2) Hippocampal neurons were fixed at DIV 10 and immunostained against vGlut1 (A-1: an excitatory neuron maker) as well as MAP2 (A-2: green) and <t>βII</t> <t>spectrin</t> (A-2: red). The specificity of vGlut1 staining is evident from the fluorescent intensity contrast observed between the arrow-marked neurons and other neurons in the field of view (A-1). (B-1 and B-2) Similar to A-1 and A-2 except that the neurons were immunostained against parvalbumin, an inhibitory neuron marker. (C-1 and C-2) Similar to A-1 and A-2 except the neurons were immunostained against GAD2, an inhibitory neuron marker. (Scale bar: 50 µm.)
Rabbit Anti Eyfp, supplied by Biorbyt, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/guinea+pig+polyclonal+anti-gad2/EYFP+antibody/bio_rxiv__2022__07__04__498712-114-70-74
Average 93 stars, based on 1 article reviews
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anti gad2  (Cell Signaling Technology Inc)
93
Cell Signaling Technology Inc anti gad2
Immunolabeling of excitatory and inhibitory neurons with specific markers. (A-1 and A-2) Hippocampal neurons were fixed at DIV 10 and immunostained against vGlut1 (A-1: an excitatory neuron maker) as well as MAP2 (A-2: green) and <t>βII</t> <t>spectrin</t> (A-2: red). The specificity of vGlut1 staining is evident from the fluorescent intensity contrast observed between the arrow-marked neurons and other neurons in the field of view (A-1). (B-1 and B-2) Similar to A-1 and A-2 except that the neurons were immunostained against parvalbumin, an inhibitory neuron marker. (C-1 and C-2) Similar to A-1 and A-2 except the neurons were immunostained against GAD2, an inhibitory neuron marker. (Scale bar: 50 µm.)
Anti Gad2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/guinea+pig+polyclonal+anti-gad2/GAD2+Antibody/pm31371378-256-77-81
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anti gad2 - by Bioz Stars, 2026-09
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rabbit anti c fos  (Cell Signaling Technology Inc)
98
Cell Signaling Technology Inc rabbit anti c fos
Immunolabeling of excitatory and inhibitory neurons with specific markers. (A-1 and A-2) Hippocampal neurons were fixed at DIV 10 and immunostained against vGlut1 (A-1: an excitatory neuron maker) as well as MAP2 (A-2: green) and <t>βII</t> <t>spectrin</t> (A-2: red). The specificity of vGlut1 staining is evident from the fluorescent intensity contrast observed between the arrow-marked neurons and other neurons in the field of view (A-1). (B-1 and B-2) Similar to A-1 and A-2 except that the neurons were immunostained against parvalbumin, an inhibitory neuron marker. (C-1 and C-2) Similar to A-1 and A-2 except the neurons were immunostained against GAD2, an inhibitory neuron marker. (Scale bar: 50 µm.)
Rabbit Anti C Fos, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/guinea+pig+polyclonal+anti-gad2/c-Fos+Rabbit+mAb/pmc12076219-234-16-19
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Image Search Results


Cortical deletion of SERT results in synaptic hyperinnervation of the DRN. a Diagram summarizing the excitatory glutamate and inhibitory GABAergic synaptic inputs received by the dorsal raphe nucleus (DRN) neurons. Synaptic inputs can be selectively identified in array tomography by the presence of specific synaptic markers including the vesicular glutamate transporter type 1 and 2 (VGLUT1 and VGLUT2, respectively) and the enzyme responsible for GABA synthesis, the glutamate decarboxylase 2 (GAD2). The prefrontal cortex (PFC), lateral habenula (LHb), laterodorsal tegmental nucleus (LDTg), ventral tegmental area (VTA), substantia nigra (SN), rostromedial tegmental nucleus (RMTg), periaqueductal gray (PAG), and hypothalamus (Hyp), have been noted as the main synaptic inputs to the DRN [ , , ]. b Immunolabeling against the 5-HT biosynthetic enzyme tryptophan hydroxylase (TPH) illustrating the distribution of 5-HT neurons in the midbrain DRN. The bracketed area shows the sampling region in the midline DRN used for array tomography quantitative analyses (at P28). c Array tomography projection image of three serial-ultrathin 70-nm-thick sections of the DRN immunolabeled against VGLUT1 (green) and synapsin (red) to specifically identify cortical synaptic boutons. The arrows indicate double-labeled boutons in control and SERT-KO mice. d-e Quantitative analysis of cortical glutamate synaptic boutons (VGLUT1+) ( d ), and subcortical glutamate (VGLUT2+) and GABAergic (GAD2+) synaptic boutons ( e ) in the DRN of control and SERT-KO mice (4 mice/genotype; for VGLUT1/Synapsin pairs: F 1,6 = 36.45, * p < 0.001; for VGLUT2/Synapsin pairs: F 1,6 = 0.24, p = 0.64; and for GAD2/Synapsin pairs: F 1,6 = 0.34, p = 0.58). f Analysis of cortical synaptic boutons in the DRN after fluoxetine-treatment (FLX) during the postnatal critical period (P2-14) (5 mice/group; F 1,8 = 9.94, * p < 0.02). g-h Density of cortical synaptic boutons in the DRN after conditional SERT invalidation using Emx1b Cre/+ :Sert fl/fl mice (SERT-KO CTX ) ( g ) and Pet1 Cre :Sert fl/fl mice (SERT-KO Raphe ) ( h ). ( g): 5 mice/genotype (Welch’s statistic = 11.20, * p < 0.03); ( h ): 3–4 mice/genotype (F 1,5 = 3.76, p = 0.11). i Cortical synaptic boutons in the mediodorsal thalamic nucleus (MD) of control and SERT-KO mice (3 mice/genotype; F 1,4 = 18.16, * p < 0.02). One-way ANOVA ( d, e, f, h, i ), and Welch’s t test ( g ). Error bars represent SEM

Journal: Molecular Psychiatry

Article Title: SSRIs target prefrontal to raphe circuits during development modulating synaptic connectivity and emotional behavior

doi: 10.1038/s41380-018-0260-9

Figure Lengend Snippet: Cortical deletion of SERT results in synaptic hyperinnervation of the DRN. a Diagram summarizing the excitatory glutamate and inhibitory GABAergic synaptic inputs received by the dorsal raphe nucleus (DRN) neurons. Synaptic inputs can be selectively identified in array tomography by the presence of specific synaptic markers including the vesicular glutamate transporter type 1 and 2 (VGLUT1 and VGLUT2, respectively) and the enzyme responsible for GABA synthesis, the glutamate decarboxylase 2 (GAD2). The prefrontal cortex (PFC), lateral habenula (LHb), laterodorsal tegmental nucleus (LDTg), ventral tegmental area (VTA), substantia nigra (SN), rostromedial tegmental nucleus (RMTg), periaqueductal gray (PAG), and hypothalamus (Hyp), have been noted as the main synaptic inputs to the DRN [ , , ]. b Immunolabeling against the 5-HT biosynthetic enzyme tryptophan hydroxylase (TPH) illustrating the distribution of 5-HT neurons in the midbrain DRN. The bracketed area shows the sampling region in the midline DRN used for array tomography quantitative analyses (at P28). c Array tomography projection image of three serial-ultrathin 70-nm-thick sections of the DRN immunolabeled against VGLUT1 (green) and synapsin (red) to specifically identify cortical synaptic boutons. The arrows indicate double-labeled boutons in control and SERT-KO mice. d-e Quantitative analysis of cortical glutamate synaptic boutons (VGLUT1+) ( d ), and subcortical glutamate (VGLUT2+) and GABAergic (GAD2+) synaptic boutons ( e ) in the DRN of control and SERT-KO mice (4 mice/genotype; for VGLUT1/Synapsin pairs: F 1,6 = 36.45, * p < 0.001; for VGLUT2/Synapsin pairs: F 1,6 = 0.24, p = 0.64; and for GAD2/Synapsin pairs: F 1,6 = 0.34, p = 0.58). f Analysis of cortical synaptic boutons in the DRN after fluoxetine-treatment (FLX) during the postnatal critical period (P2-14) (5 mice/group; F 1,8 = 9.94, * p < 0.02). g-h Density of cortical synaptic boutons in the DRN after conditional SERT invalidation using Emx1b Cre/+ :Sert fl/fl mice (SERT-KO CTX ) ( g ) and Pet1 Cre :Sert fl/fl mice (SERT-KO Raphe ) ( h ). ( g): 5 mice/genotype (Welch’s statistic = 11.20, * p < 0.03); ( h ): 3–4 mice/genotype (F 1,5 = 3.76, p = 0.11). i Cortical synaptic boutons in the mediodorsal thalamic nucleus (MD) of control and SERT-KO mice (3 mice/genotype; F 1,4 = 18.16, * p < 0.02). One-way ANOVA ( d, e, f, h, i ), and Welch’s t test ( g ). Error bars represent SEM

Article Snippet: The following antibodies were used: anti-DsRed (rabbit, 1:400; Clontech, USA; #632496) [ ], anti-VGLUT1 (guinea pig, 1:1000; Millipore, USA; AB5905), anti-VGLUT2 (guinea pig, 1:1000; Millipore, USA; AB2251), and anti-glutamate decarboxylase 65 (GAD2) (rabbit, 1:200; Cell Signaling Technology, USA; #5843) [ , ].

Techniques: Tomography, Immunolabeling, Sampling, Labeling, Control

Immunolabeling of excitatory and inhibitory neurons with specific markers. (A-1 and A-2) Hippocampal neurons were fixed at DIV 10 and immunostained against vGlut1 (A-1: an excitatory neuron maker) as well as MAP2 (A-2: green) and βII spectrin (A-2: red). The specificity of vGlut1 staining is evident from the fluorescent intensity contrast observed between the arrow-marked neurons and other neurons in the field of view (A-1). (B-1 and B-2) Similar to A-1 and A-2 except that the neurons were immunostained against parvalbumin, an inhibitory neuron marker. (C-1 and C-2) Similar to A-1 and A-2 except the neurons were immunostained against GAD2, an inhibitory neuron marker. (Scale bar: 50 µm.)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

doi: 10.1073/pnas.1605707113

Figure Lengend Snippet: Immunolabeling of excitatory and inhibitory neurons with specific markers. (A-1 and A-2) Hippocampal neurons were fixed at DIV 10 and immunostained against vGlut1 (A-1: an excitatory neuron maker) as well as MAP2 (A-2: green) and βII spectrin (A-2: red). The specificity of vGlut1 staining is evident from the fluorescent intensity contrast observed between the arrow-marked neurons and other neurons in the field of view (A-1). (B-1 and B-2) Similar to A-1 and A-2 except that the neurons were immunostained against parvalbumin, an inhibitory neuron marker. (C-1 and C-2) Similar to A-1 and A-2 except the neurons were immunostained against GAD2, an inhibitory neuron marker. (Scale bar: 50 µm.)

Article Snippet: The following primary antibodies were used in this study: guinea pig anti-MAP2 antibody (Synaptic Systems, 188002); mouse anti-βII spectrin antibody (BD Biosciences, 612563); rabbit anti-GFP antibody (Abcam, ab290); guinea pig anti-vGlut1 antibody (Synaptic Systems, 135304); rabbit anti-calbindin antibody (Swant, CB38); rabbit anti-tyrosine hydroxylase antibody (Abcam, ab6211); rabbit anti-GAD2 antibody (Synaptic Systems, 198103); rabbit anti-GAD1/GAD2 antibody (Abcam ab11070); goat anti-parvalbumin antibody (Santa Cruz, sc-7449); rabbit anti-parvalbumin antibody (Abcam, ab11427); rabbit anti-metabotropic glutamate receptor antibody (Abcam, ab6438); rat anti-MBP antibody (Millipore, Mab386); rabbit anti-OMP antibody (Santa Cruz, sc-67219); rabbit anti-NG2 antibody (Millipore, AB5320); rabbit anti-CD11b (Novus Biologicals, NB110-89474); rabbit anti-S100b (Abcam, ab41548); rabbit anti-GFAP (Encor, RPCA-GFAP), and rabbit anti-NeuN (Millipore, MABN140).

Techniques: Immunolabeling, Staining, Marker

The MPS structure is present in both excitatory and inhibitory neurons from mouse cortex, midbrain, and hippocampus. Cultured neurons were immunostained with vGlut1 or GAD2 to label the excitatory and inhibitory neurons, respectively. (A) Reconstructed conventional image of a vGlut1+ cortical neuron shown together with 3D STORM images of βII spectrin in an axonal (A-1) and dendritic (A-2) region. The axonal (A-1) and dendritic (A-2) regions correspond to the regions indicated by arrows in A. Dotted lines in A-2 indicate patches of periodic pattern in the dendritic shaft. In 3D STORM images, localizations at different z values are depicted in different colors. (B, B-1, and B-2) Similar to A, A-1, and A-2 but for a GAD2+ cortical neuron. (C) Averaged autocorrelation functions calculated from multiple, randomly selected axonal (black) and dendritic (red) regions in neurons cultured from cortex (n = 18 for axons and n = 9 for dendrites of GAD2+ neurons; n = 9 for axons and n = 8 for dendrites of vGlut1+ neurons), midbrain (n = 8 for axons and n = 7 for dendrites of GAD2+ neurons; n = 13 for axons and n = 11 for dendrites of vGlut1+ neurons), and hippocampus (n = 6 for axons and n = 8 for dendrites of GAD2+ neurons; n = 8 for axons and n = 9 for dendrites of vGlut1+ neurons), respectively. (Scale bar: 20 µm for conventional images and 2 µm for STORM images.)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

doi: 10.1073/pnas.1605707113

Figure Lengend Snippet: The MPS structure is present in both excitatory and inhibitory neurons from mouse cortex, midbrain, and hippocampus. Cultured neurons were immunostained with vGlut1 or GAD2 to label the excitatory and inhibitory neurons, respectively. (A) Reconstructed conventional image of a vGlut1+ cortical neuron shown together with 3D STORM images of βII spectrin in an axonal (A-1) and dendritic (A-2) region. The axonal (A-1) and dendritic (A-2) regions correspond to the regions indicated by arrows in A. Dotted lines in A-2 indicate patches of periodic pattern in the dendritic shaft. In 3D STORM images, localizations at different z values are depicted in different colors. (B, B-1, and B-2) Similar to A, A-1, and A-2 but for a GAD2+ cortical neuron. (C) Averaged autocorrelation functions calculated from multiple, randomly selected axonal (black) and dendritic (red) regions in neurons cultured from cortex (n = 18 for axons and n = 9 for dendrites of GAD2+ neurons; n = 9 for axons and n = 8 for dendrites of vGlut1+ neurons), midbrain (n = 8 for axons and n = 7 for dendrites of GAD2+ neurons; n = 13 for axons and n = 11 for dendrites of vGlut1+ neurons), and hippocampus (n = 6 for axons and n = 8 for dendrites of GAD2+ neurons; n = 8 for axons and n = 9 for dendrites of vGlut1+ neurons), respectively. (Scale bar: 20 µm for conventional images and 2 µm for STORM images.)

Article Snippet: The following primary antibodies were used in this study: guinea pig anti-MAP2 antibody (Synaptic Systems, 188002); mouse anti-βII spectrin antibody (BD Biosciences, 612563); rabbit anti-GFP antibody (Abcam, ab290); guinea pig anti-vGlut1 antibody (Synaptic Systems, 135304); rabbit anti-calbindin antibody (Swant, CB38); rabbit anti-tyrosine hydroxylase antibody (Abcam, ab6211); rabbit anti-GAD2 antibody (Synaptic Systems, 198103); rabbit anti-GAD1/GAD2 antibody (Abcam ab11070); goat anti-parvalbumin antibody (Santa Cruz, sc-7449); rabbit anti-parvalbumin antibody (Abcam, ab11427); rabbit anti-metabotropic glutamate receptor antibody (Abcam, ab6438); rat anti-MBP antibody (Millipore, Mab386); rabbit anti-OMP antibody (Santa Cruz, sc-67219); rabbit anti-NG2 antibody (Millipore, AB5320); rabbit anti-CD11b (Novus Biologicals, NB110-89474); rabbit anti-S100b (Abcam, ab41548); rabbit anti-GFAP (Encor, RPCA-GFAP), and rabbit anti-NeuN (Millipore, MABN140).

Techniques: Cell Culture

βII spectrin is organized into a periodic structure in both excitatory and inhibitory neurons from mouse cortex. Cultured neurons from mouse cortex were immunostained with vGlut1 or GAD2 to label the excitatory and inhibitory neurons, respectively. (A) Reconstructed conventional image of a vGlut1+ cortical neuron. (A-1 and A-2) 3D STORM images of βII spectrin in an axonal (A-1) and dendritic (A-2) region indicated by arrows in A. (Right panels) Autocorrelation functions of the selected boxed regions in the STORM images. (B, B-1, and B-2) Similar to A, A-1, and A-2 but for a GAD2+ inhibitory neuron. (Scale bar: 20 µm for conventional images and 2 µm for STORM images.)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

doi: 10.1073/pnas.1605707113

Figure Lengend Snippet: βII spectrin is organized into a periodic structure in both excitatory and inhibitory neurons from mouse cortex. Cultured neurons from mouse cortex were immunostained with vGlut1 or GAD2 to label the excitatory and inhibitory neurons, respectively. (A) Reconstructed conventional image of a vGlut1+ cortical neuron. (A-1 and A-2) 3D STORM images of βII spectrin in an axonal (A-1) and dendritic (A-2) region indicated by arrows in A. (Right panels) Autocorrelation functions of the selected boxed regions in the STORM images. (B, B-1, and B-2) Similar to A, A-1, and A-2 but for a GAD2+ inhibitory neuron. (Scale bar: 20 µm for conventional images and 2 µm for STORM images.)

Article Snippet: The following primary antibodies were used in this study: guinea pig anti-MAP2 antibody (Synaptic Systems, 188002); mouse anti-βII spectrin antibody (BD Biosciences, 612563); rabbit anti-GFP antibody (Abcam, ab290); guinea pig anti-vGlut1 antibody (Synaptic Systems, 135304); rabbit anti-calbindin antibody (Swant, CB38); rabbit anti-tyrosine hydroxylase antibody (Abcam, ab6211); rabbit anti-GAD2 antibody (Synaptic Systems, 198103); rabbit anti-GAD1/GAD2 antibody (Abcam ab11070); goat anti-parvalbumin antibody (Santa Cruz, sc-7449); rabbit anti-parvalbumin antibody (Abcam, ab11427); rabbit anti-metabotropic glutamate receptor antibody (Abcam, ab6438); rat anti-MBP antibody (Millipore, Mab386); rabbit anti-OMP antibody (Santa Cruz, sc-67219); rabbit anti-NG2 antibody (Millipore, AB5320); rabbit anti-CD11b (Novus Biologicals, NB110-89474); rabbit anti-S100b (Abcam, ab41548); rabbit anti-GFAP (Encor, RPCA-GFAP), and rabbit anti-NeuN (Millipore, MABN140).

Techniques: Cell Culture

βII spectrin is organized into a periodic structure in both excitatory and inhibitory neurons from mouse hippocampus. Cultured neurons from mouse hippocampus were immunostained with vGlut1 or GAD2 to label the excitatory and inhibitory neurons, respectively. (A) Reconstructed conventional image of a vGlut1+ hippocampal neuron. 3D STORM images of βII spectrin in an axonal (A-1) and dendritic (A-2) region indicated by arrows in A. (Right panels) Autocorrelation functions of the selected boxed regions in the A-1 and A-2 STORM images. (B, B-1, and B-2) Similar to A, A-1, and A-2, but for a GAD2+ inhibitory neuron. (Scale bar: 20 µm for conventional images and 2 µm for STORM images.)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

doi: 10.1073/pnas.1605707113

Figure Lengend Snippet: βII spectrin is organized into a periodic structure in both excitatory and inhibitory neurons from mouse hippocampus. Cultured neurons from mouse hippocampus were immunostained with vGlut1 or GAD2 to label the excitatory and inhibitory neurons, respectively. (A) Reconstructed conventional image of a vGlut1+ hippocampal neuron. 3D STORM images of βII spectrin in an axonal (A-1) and dendritic (A-2) region indicated by arrows in A. (Right panels) Autocorrelation functions of the selected boxed regions in the A-1 and A-2 STORM images. (B, B-1, and B-2) Similar to A, A-1, and A-2, but for a GAD2+ inhibitory neuron. (Scale bar: 20 µm for conventional images and 2 µm for STORM images.)

Article Snippet: The following primary antibodies were used in this study: guinea pig anti-MAP2 antibody (Synaptic Systems, 188002); mouse anti-βII spectrin antibody (BD Biosciences, 612563); rabbit anti-GFP antibody (Abcam, ab290); guinea pig anti-vGlut1 antibody (Synaptic Systems, 135304); rabbit anti-calbindin antibody (Swant, CB38); rabbit anti-tyrosine hydroxylase antibody (Abcam, ab6211); rabbit anti-GAD2 antibody (Synaptic Systems, 198103); rabbit anti-GAD1/GAD2 antibody (Abcam ab11070); goat anti-parvalbumin antibody (Santa Cruz, sc-7449); rabbit anti-parvalbumin antibody (Abcam, ab11427); rabbit anti-metabotropic glutamate receptor antibody (Abcam, ab6438); rat anti-MBP antibody (Millipore, Mab386); rabbit anti-OMP antibody (Santa Cruz, sc-67219); rabbit anti-NG2 antibody (Millipore, AB5320); rabbit anti-CD11b (Novus Biologicals, NB110-89474); rabbit anti-S100b (Abcam, ab41548); rabbit anti-GFAP (Encor, RPCA-GFAP), and rabbit anti-NeuN (Millipore, MABN140).

Techniques: Cell Culture

βII spectrin is organized into a periodic structure in both excitatory and inhibitory neurons from mouse midbrain. Cultured neurons from mouse midbrain were immunostained with vGlut1 or GAD2 to label the excitatory and inhibitory neurons, respectively. (A) Reconstructed conventional image of a vGlut1+ midbrain neuron. (A-1 and A-2) 3D STORM images of βII spectrin in an axonal (A-1) and dendritic (A-2) region indicated by arrows in A. (Right panels) Autocorrelation functions of the selected boxed regions in the A-1 and A-2 STORM images. (B, B-1, and B-2) Similar to A, A-1, and A-2 but for a GAD2+ inhibitory neuron. (Scale bar: 20 µm for conventional images and 2 µm for STORM images.)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

doi: 10.1073/pnas.1605707113

Figure Lengend Snippet: βII spectrin is organized into a periodic structure in both excitatory and inhibitory neurons from mouse midbrain. Cultured neurons from mouse midbrain were immunostained with vGlut1 or GAD2 to label the excitatory and inhibitory neurons, respectively. (A) Reconstructed conventional image of a vGlut1+ midbrain neuron. (A-1 and A-2) 3D STORM images of βII spectrin in an axonal (A-1) and dendritic (A-2) region indicated by arrows in A. (Right panels) Autocorrelation functions of the selected boxed regions in the A-1 and A-2 STORM images. (B, B-1, and B-2) Similar to A, A-1, and A-2 but for a GAD2+ inhibitory neuron. (Scale bar: 20 µm for conventional images and 2 µm for STORM images.)

Article Snippet: The following primary antibodies were used in this study: guinea pig anti-MAP2 antibody (Synaptic Systems, 188002); mouse anti-βII spectrin antibody (BD Biosciences, 612563); rabbit anti-GFP antibody (Abcam, ab290); guinea pig anti-vGlut1 antibody (Synaptic Systems, 135304); rabbit anti-calbindin antibody (Swant, CB38); rabbit anti-tyrosine hydroxylase antibody (Abcam, ab6211); rabbit anti-GAD2 antibody (Synaptic Systems, 198103); rabbit anti-GAD1/GAD2 antibody (Abcam ab11070); goat anti-parvalbumin antibody (Santa Cruz, sc-7449); rabbit anti-parvalbumin antibody (Abcam, ab11427); rabbit anti-metabotropic glutamate receptor antibody (Abcam, ab6438); rat anti-MBP antibody (Millipore, Mab386); rabbit anti-OMP antibody (Santa Cruz, sc-67219); rabbit anti-NG2 antibody (Millipore, AB5320); rabbit anti-CD11b (Novus Biologicals, NB110-89474); rabbit anti-S100b (Abcam, ab41548); rabbit anti-GFAP (Encor, RPCA-GFAP), and rabbit anti-NeuN (Millipore, MABN140).

Techniques: Cell Culture

Identification of some specific subtypes of neurons from mouse central nervous system using specific markers. Neurons from different regions of the central nervous system were cultured for at least 10 days in vitro and then immunostained with a specific neuronal marker (A-1, B-1, and C-1) as well as MAP2 (green; A-2, B-2, and C-2) and βII spectrin (red; A-2, B-2, and C-2). (A-1 and A-2) Golgi cells from cerebellum are marked by metabotropic glutamate receptor staining. (B-1 and B-2) Purkinje cells from cerebellum are marked by calbindin staining. (C-1 and C-2) Dopaminergic neurons from midbrain are marked by tyrosine hydroxylase staining. (Scale bar: 50 µm.)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

doi: 10.1073/pnas.1605707113

Figure Lengend Snippet: Identification of some specific subtypes of neurons from mouse central nervous system using specific markers. Neurons from different regions of the central nervous system were cultured for at least 10 days in vitro and then immunostained with a specific neuronal marker (A-1, B-1, and C-1) as well as MAP2 (green; A-2, B-2, and C-2) and βII spectrin (red; A-2, B-2, and C-2). (A-1 and A-2) Golgi cells from cerebellum are marked by metabotropic glutamate receptor staining. (B-1 and B-2) Purkinje cells from cerebellum are marked by calbindin staining. (C-1 and C-2) Dopaminergic neurons from midbrain are marked by tyrosine hydroxylase staining. (Scale bar: 50 µm.)

Article Snippet: The following primary antibodies were used in this study: guinea pig anti-MAP2 antibody (Synaptic Systems, 188002); mouse anti-βII spectrin antibody (BD Biosciences, 612563); rabbit anti-GFP antibody (Abcam, ab290); guinea pig anti-vGlut1 antibody (Synaptic Systems, 135304); rabbit anti-calbindin antibody (Swant, CB38); rabbit anti-tyrosine hydroxylase antibody (Abcam, ab6211); rabbit anti-GAD2 antibody (Synaptic Systems, 198103); rabbit anti-GAD1/GAD2 antibody (Abcam ab11070); goat anti-parvalbumin antibody (Santa Cruz, sc-7449); rabbit anti-parvalbumin antibody (Abcam, ab11427); rabbit anti-metabotropic glutamate receptor antibody (Abcam, ab6438); rat anti-MBP antibody (Millipore, Mab386); rabbit anti-OMP antibody (Santa Cruz, sc-67219); rabbit anti-NG2 antibody (Millipore, AB5320); rabbit anti-CD11b (Novus Biologicals, NB110-89474); rabbit anti-S100b (Abcam, ab41548); rabbit anti-GFAP (Encor, RPCA-GFAP), and rabbit anti-NeuN (Millipore, MABN140).

Techniques: Cell Culture, In Vitro, Marker, Staining

MPS is present in multiple subtypes of inhibitory and excitatory neurons in the mouse central nervous system. (A–I) Representative STORM images of βII spectrin for a typical axonal region (Left panels in A–I) and averaged autocorrelation functions from multiple axons (Right panels A–I) for cultured cerebellar granule cells (A; n = 7 for averaged autocorrelation calculation), Pontine nuclei neurons (B; n = 7), dopaminergic neurons (C; n = 9), olfactory neurons (D; n = 16), Parvalbumin neurons from cortex (E; n = 9), Parvalbumin neurons from midbrain (F; n = 6), Parvalbumin neurons from hippocampus (G; n = 13), Golgi cells (H; n = 8), and Purkinje cells (I; n = 7) from cerebellum. (J and K) Averaged autocorrelation amplitudes of axons (blue) versus dendrites (red) for the neuronal subtypes we imaged in the mouse central nervous system. Error bars are SEM. (Scale bar: 2 µm.)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

doi: 10.1073/pnas.1605707113

Figure Lengend Snippet: MPS is present in multiple subtypes of inhibitory and excitatory neurons in the mouse central nervous system. (A–I) Representative STORM images of βII spectrin for a typical axonal region (Left panels in A–I) and averaged autocorrelation functions from multiple axons (Right panels A–I) for cultured cerebellar granule cells (A; n = 7 for averaged autocorrelation calculation), Pontine nuclei neurons (B; n = 7), dopaminergic neurons (C; n = 9), olfactory neurons (D; n = 16), Parvalbumin neurons from cortex (E; n = 9), Parvalbumin neurons from midbrain (F; n = 6), Parvalbumin neurons from hippocampus (G; n = 13), Golgi cells (H; n = 8), and Purkinje cells (I; n = 7) from cerebellum. (J and K) Averaged autocorrelation amplitudes of axons (blue) versus dendrites (red) for the neuronal subtypes we imaged in the mouse central nervous system. Error bars are SEM. (Scale bar: 2 µm.)

Article Snippet: The following primary antibodies were used in this study: guinea pig anti-MAP2 antibody (Synaptic Systems, 188002); mouse anti-βII spectrin antibody (BD Biosciences, 612563); rabbit anti-GFP antibody (Abcam, ab290); guinea pig anti-vGlut1 antibody (Synaptic Systems, 135304); rabbit anti-calbindin antibody (Swant, CB38); rabbit anti-tyrosine hydroxylase antibody (Abcam, ab6211); rabbit anti-GAD2 antibody (Synaptic Systems, 198103); rabbit anti-GAD1/GAD2 antibody (Abcam ab11070); goat anti-parvalbumin antibody (Santa Cruz, sc-7449); rabbit anti-parvalbumin antibody (Abcam, ab11427); rabbit anti-metabotropic glutamate receptor antibody (Abcam, ab6438); rat anti-MBP antibody (Millipore, Mab386); rabbit anti-OMP antibody (Santa Cruz, sc-67219); rabbit anti-NG2 antibody (Millipore, AB5320); rabbit anti-CD11b (Novus Biologicals, NB110-89474); rabbit anti-S100b (Abcam, ab41548); rabbit anti-GFAP (Encor, RPCA-GFAP), and rabbit anti-NeuN (Millipore, MABN140).

Techniques: Cell Culture

In vitro myelination does not affect the formation of the MPS structure. Hippocampal neurons were cultured for 2 wk before adding the glia cells. After coculturing for 6 wk, the cells were fixed and immunostained with βII spectrin (green) and MBP (myelination marker, magenta). (A) Conventional image of neurons showing a myelinated axon (shown in magenta). (Scale bar: 10 µm.) (B) 3D STORM image of βII spectrin in the same area as in A. The myelin sheath was shown in magenta and imaged at the conventional resolution. βII spectrin adopts a highly periodic structure in both the myelinated and unmyelinated regions along the axon. (Scale bar: 10 µm.) (C) Zoom-in 3D STORM image of βII spectrin from the two boxed regions from B). (Scale bar: 2 µm.) (D) Averaged autocorrelation functions of βII-spectrin distribution calculated from multiple MBP+ and MBP− axons (n = 7 each).

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

doi: 10.1073/pnas.1605707113

Figure Lengend Snippet: In vitro myelination does not affect the formation of the MPS structure. Hippocampal neurons were cultured for 2 wk before adding the glia cells. After coculturing for 6 wk, the cells were fixed and immunostained with βII spectrin (green) and MBP (myelination marker, magenta). (A) Conventional image of neurons showing a myelinated axon (shown in magenta). (Scale bar: 10 µm.) (B) 3D STORM image of βII spectrin in the same area as in A. The myelin sheath was shown in magenta and imaged at the conventional resolution. βII spectrin adopts a highly periodic structure in both the myelinated and unmyelinated regions along the axon. (Scale bar: 10 µm.) (C) Zoom-in 3D STORM image of βII spectrin from the two boxed regions from B). (Scale bar: 2 µm.) (D) Averaged autocorrelation functions of βII-spectrin distribution calculated from multiple MBP+ and MBP− axons (n = 7 each).

Article Snippet: The following primary antibodies were used in this study: guinea pig anti-MAP2 antibody (Synaptic Systems, 188002); mouse anti-βII spectrin antibody (BD Biosciences, 612563); rabbit anti-GFP antibody (Abcam, ab290); guinea pig anti-vGlut1 antibody (Synaptic Systems, 135304); rabbit anti-calbindin antibody (Swant, CB38); rabbit anti-tyrosine hydroxylase antibody (Abcam, ab6211); rabbit anti-GAD2 antibody (Synaptic Systems, 198103); rabbit anti-GAD1/GAD2 antibody (Abcam ab11070); goat anti-parvalbumin antibody (Santa Cruz, sc-7449); rabbit anti-parvalbumin antibody (Abcam, ab11427); rabbit anti-metabotropic glutamate receptor antibody (Abcam, ab6438); rat anti-MBP antibody (Millipore, Mab386); rabbit anti-OMP antibody (Santa Cruz, sc-67219); rabbit anti-NG2 antibody (Millipore, AB5320); rabbit anti-CD11b (Novus Biologicals, NB110-89474); rabbit anti-S100b (Abcam, ab41548); rabbit anti-GFAP (Encor, RPCA-GFAP), and rabbit anti-NeuN (Millipore, MABN140).

Techniques: In Vitro, Cell Culture, Marker

MPS is present in mouse peripheral sensory and motor axons. (A) Representative STORM images of βII spectrin for a typical axonal region (Left) and averaged autocorrelation functions from multiple axons (Right, n = 12) for mES-derived motor neurons. (B) Same as in A but for dendrites in mES-derived motor neurons (n = 11). (C and D) Same as A but for axons in DRG neurons (C, explants, n = 13; D, dissociated DRG culture, n = 10). (Scale bar: 2 µm.)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

doi: 10.1073/pnas.1605707113

Figure Lengend Snippet: MPS is present in mouse peripheral sensory and motor axons. (A) Representative STORM images of βII spectrin for a typical axonal region (Left) and averaged autocorrelation functions from multiple axons (Right, n = 12) for mES-derived motor neurons. (B) Same as in A but for dendrites in mES-derived motor neurons (n = 11). (C and D) Same as A but for axons in DRG neurons (C, explants, n = 13; D, dissociated DRG culture, n = 10). (Scale bar: 2 µm.)

Article Snippet: The following primary antibodies were used in this study: guinea pig anti-MAP2 antibody (Synaptic Systems, 188002); mouse anti-βII spectrin antibody (BD Biosciences, 612563); rabbit anti-GFP antibody (Abcam, ab290); guinea pig anti-vGlut1 antibody (Synaptic Systems, 135304); rabbit anti-calbindin antibody (Swant, CB38); rabbit anti-tyrosine hydroxylase antibody (Abcam, ab6211); rabbit anti-GAD2 antibody (Synaptic Systems, 198103); rabbit anti-GAD1/GAD2 antibody (Abcam ab11070); goat anti-parvalbumin antibody (Santa Cruz, sc-7449); rabbit anti-parvalbumin antibody (Abcam, ab11427); rabbit anti-metabotropic glutamate receptor antibody (Abcam, ab6438); rat anti-MBP antibody (Millipore, Mab386); rabbit anti-OMP antibody (Santa Cruz, sc-67219); rabbit anti-NG2 antibody (Millipore, AB5320); rabbit anti-CD11b (Novus Biologicals, NB110-89474); rabbit anti-S100b (Abcam, ab41548); rabbit anti-GFAP (Encor, RPCA-GFAP), and rabbit anti-NeuN (Millipore, MABN140).

Techniques: Derivative Assay

The distribution of βII spectrin in axonal boutons and dendritic spines. (A) Representative STORM images of βII spectrin in axons that contain presynaptic boutons marked by a presynaptic marker, Bassoon (magenta). βII spectrin is labeled by expressing GFP-βII spectrin (or GFP-αII spectrin) in a sparse subset of cultured neurons, and hence some bassoon-positive regions do not overlap with the GFP-positive axon. (Top and Middle) Examples of bassoon-positive presynaptic boutons (indicated by arrow) within which the periodic pattern of spectrin is disrupted. (Bottom) An example of bassoon-positive presynaptic bouton (indicated by arrow) within which the period pattern of spectrin is not disrupted. (B) Bar graph depicting relative fraction of boutons in which the periodic structure is disrupted or undisrupted. (C) The dendritic region of a cultured mouse hippocampal neuron immunostained with MAP2, a dendrite marker. (C-1 and C-2) STORM images of two selected regions showing periodic patterns of immunolabeled endogenous βII spectrin in spine necks (Left panels) with autocorrelation functions (Right panels) calculated from the boxed spine neck region (white boxes) and dendritic shaft (yellow boxes). (Scale bar: 10 µm for conventional images and 1 µm for STORM images.)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

doi: 10.1073/pnas.1605707113

Figure Lengend Snippet: The distribution of βII spectrin in axonal boutons and dendritic spines. (A) Representative STORM images of βII spectrin in axons that contain presynaptic boutons marked by a presynaptic marker, Bassoon (magenta). βII spectrin is labeled by expressing GFP-βII spectrin (or GFP-αII spectrin) in a sparse subset of cultured neurons, and hence some bassoon-positive regions do not overlap with the GFP-positive axon. (Top and Middle) Examples of bassoon-positive presynaptic boutons (indicated by arrow) within which the periodic pattern of spectrin is disrupted. (Bottom) An example of bassoon-positive presynaptic bouton (indicated by arrow) within which the period pattern of spectrin is not disrupted. (B) Bar graph depicting relative fraction of boutons in which the periodic structure is disrupted or undisrupted. (C) The dendritic region of a cultured mouse hippocampal neuron immunostained with MAP2, a dendrite marker. (C-1 and C-2) STORM images of two selected regions showing periodic patterns of immunolabeled endogenous βII spectrin in spine necks (Left panels) with autocorrelation functions (Right panels) calculated from the boxed spine neck region (white boxes) and dendritic shaft (yellow boxes). (Scale bar: 10 µm for conventional images and 1 µm for STORM images.)

Article Snippet: The following primary antibodies were used in this study: guinea pig anti-MAP2 antibody (Synaptic Systems, 188002); mouse anti-βII spectrin antibody (BD Biosciences, 612563); rabbit anti-GFP antibody (Abcam, ab290); guinea pig anti-vGlut1 antibody (Synaptic Systems, 135304); rabbit anti-calbindin antibody (Swant, CB38); rabbit anti-tyrosine hydroxylase antibody (Abcam, ab6211); rabbit anti-GAD2 antibody (Synaptic Systems, 198103); rabbit anti-GAD1/GAD2 antibody (Abcam ab11070); goat anti-parvalbumin antibody (Santa Cruz, sc-7449); rabbit anti-parvalbumin antibody (Abcam, ab11427); rabbit anti-metabotropic glutamate receptor antibody (Abcam, ab6438); rat anti-MBP antibody (Millipore, Mab386); rabbit anti-OMP antibody (Santa Cruz, sc-67219); rabbit anti-NG2 antibody (Millipore, AB5320); rabbit anti-CD11b (Novus Biologicals, NB110-89474); rabbit anti-S100b (Abcam, ab41548); rabbit anti-GFAP (Encor, RPCA-GFAP), and rabbit anti-NeuN (Millipore, MABN140).

Techniques: Marker, Labeling, Expressing, Cell Culture, Immunolabeling

Sparse presence of the periodic βII spectrin structure in glial cell processes. (A) An astrocyte stained for the astrocyte marker GFAP (green) and βII spectrin (red). (A-1 and A-2) STORM images of βII spectrin from two processes of that astrocyte (indicated by arrows) displaying relatively periodic (A-1) or irregular (A-2) spectrin distribution. (Right panels) Autocorrelation analysis of the boxed regions. (B) Averaged autocorrelation functions (blue) calculated from multiple randomly selected processes of astrocytes, microglia cells, NG2 glia, and Schwann cells (n = 10 for astrocytes, n = 9 for microglia, n = 8 for NG2 glia, n = 9 for Schwann cells). For comparison, the dotted gray and red curves show the averaged autocorrelation functions of βII spectrin distribution in axons and dendrites of cortical vGlut1+ neurons, respectively (reproduced from Fig. 1C). (Scale bar: 20 µm for conventional image and 1 µm for STORM images.)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

doi: 10.1073/pnas.1605707113

Figure Lengend Snippet: Sparse presence of the periodic βII spectrin structure in glial cell processes. (A) An astrocyte stained for the astrocyte marker GFAP (green) and βII spectrin (red). (A-1 and A-2) STORM images of βII spectrin from two processes of that astrocyte (indicated by arrows) displaying relatively periodic (A-1) or irregular (A-2) spectrin distribution. (Right panels) Autocorrelation analysis of the boxed regions. (B) Averaged autocorrelation functions (blue) calculated from multiple randomly selected processes of astrocytes, microglia cells, NG2 glia, and Schwann cells (n = 10 for astrocytes, n = 9 for microglia, n = 8 for NG2 glia, n = 9 for Schwann cells). For comparison, the dotted gray and red curves show the averaged autocorrelation functions of βII spectrin distribution in axons and dendrites of cortical vGlut1+ neurons, respectively (reproduced from Fig. 1C). (Scale bar: 20 µm for conventional image and 1 µm for STORM images.)

Article Snippet: The following primary antibodies were used in this study: guinea pig anti-MAP2 antibody (Synaptic Systems, 188002); mouse anti-βII spectrin antibody (BD Biosciences, 612563); rabbit anti-GFP antibody (Abcam, ab290); guinea pig anti-vGlut1 antibody (Synaptic Systems, 135304); rabbit anti-calbindin antibody (Swant, CB38); rabbit anti-tyrosine hydroxylase antibody (Abcam, ab6211); rabbit anti-GAD2 antibody (Synaptic Systems, 198103); rabbit anti-GAD1/GAD2 antibody (Abcam ab11070); goat anti-parvalbumin antibody (Santa Cruz, sc-7449); rabbit anti-parvalbumin antibody (Abcam, ab11427); rabbit anti-metabotropic glutamate receptor antibody (Abcam, ab6438); rat anti-MBP antibody (Millipore, Mab386); rabbit anti-OMP antibody (Santa Cruz, sc-67219); rabbit anti-NG2 antibody (Millipore, AB5320); rabbit anti-CD11b (Novus Biologicals, NB110-89474); rabbit anti-S100b (Abcam, ab41548); rabbit anti-GFAP (Encor, RPCA-GFAP), and rabbit anti-NeuN (Millipore, MABN140).

Techniques: Staining, Marker

The distribution of βII spectrin in the processes of microglia, NG2 glia and Schwann cells. (A) Conventional image of neurons and microglial cells stained with the microglia marker CD11b (green) and MAP2 (blue). The microglial cells in the field of view appear green. (A-1 and A-2) Selected 3D STORM images of βII spectrin from the process of a microglial cell (indicated by arrows in A) displaying relatively periodic (A-1) or irregular (A-2) spectrin distribution. (Right) Autocorrelation functions from the boxed regions. (B, B-1, and B-2) Similar to A, A-1, and A-2 but for a NG2+ glial cell labeled by NG2 (green). MAP2 staining is shown in blue. (C, C-1, and C-2) Similar to A, A-1, and A-2 but for Schwann cells labeled with the Schwann cell marker S100b (green). βII-spectrin staining is shown in red. (Scale bar: 20 µm for conventional images and 1 µm for STORM images.)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

doi: 10.1073/pnas.1605707113

Figure Lengend Snippet: The distribution of βII spectrin in the processes of microglia, NG2 glia and Schwann cells. (A) Conventional image of neurons and microglial cells stained with the microglia marker CD11b (green) and MAP2 (blue). The microglial cells in the field of view appear green. (A-1 and A-2) Selected 3D STORM images of βII spectrin from the process of a microglial cell (indicated by arrows in A) displaying relatively periodic (A-1) or irregular (A-2) spectrin distribution. (Right) Autocorrelation functions from the boxed regions. (B, B-1, and B-2) Similar to A, A-1, and A-2 but for a NG2+ glial cell labeled by NG2 (green). MAP2 staining is shown in blue. (C, C-1, and C-2) Similar to A, A-1, and A-2 but for Schwann cells labeled with the Schwann cell marker S100b (green). βII-spectrin staining is shown in red. (Scale bar: 20 µm for conventional images and 1 µm for STORM images.)

Article Snippet: The following primary antibodies were used in this study: guinea pig anti-MAP2 antibody (Synaptic Systems, 188002); mouse anti-βII spectrin antibody (BD Biosciences, 612563); rabbit anti-GFP antibody (Abcam, ab290); guinea pig anti-vGlut1 antibody (Synaptic Systems, 135304); rabbit anti-calbindin antibody (Swant, CB38); rabbit anti-tyrosine hydroxylase antibody (Abcam, ab6211); rabbit anti-GAD2 antibody (Synaptic Systems, 198103); rabbit anti-GAD1/GAD2 antibody (Abcam ab11070); goat anti-parvalbumin antibody (Santa Cruz, sc-7449); rabbit anti-parvalbumin antibody (Abcam, ab11427); rabbit anti-metabotropic glutamate receptor antibody (Abcam, ab6438); rat anti-MBP antibody (Millipore, Mab386); rabbit anti-OMP antibody (Santa Cruz, sc-67219); rabbit anti-NG2 antibody (Millipore, AB5320); rabbit anti-CD11b (Novus Biologicals, NB110-89474); rabbit anti-S100b (Abcam, ab41548); rabbit anti-GFAP (Encor, RPCA-GFAP), and rabbit anti-NeuN (Millipore, MABN140).

Techniques: Staining, Marker, Labeling

Structural domain organization of α and β spectrin across different species. The structural domains of αII and βII spectrin homologs, including spectrin domain (blue), SH3 domain (yellow), calcium-binding EF band domain (red), PH domain (magenta), and calponin homology (CH, green) domain from C. elegans, Drosophila, G. gallus, Mus musculus, and H. sapiens. Also shown are the lengths of these proteins in terms of the number of amino acids (aa).

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

doi: 10.1073/pnas.1605707113

Figure Lengend Snippet: Structural domain organization of α and β spectrin across different species. The structural domains of αII and βII spectrin homologs, including spectrin domain (blue), SH3 domain (yellow), calcium-binding EF band domain (red), PH domain (magenta), and calponin homology (CH, green) domain from C. elegans, Drosophila, G. gallus, Mus musculus, and H. sapiens. Also shown are the lengths of these proteins in terms of the number of amino acids (aa).

Article Snippet: The following primary antibodies were used in this study: guinea pig anti-MAP2 antibody (Synaptic Systems, 188002); mouse anti-βII spectrin antibody (BD Biosciences, 612563); rabbit anti-GFP antibody (Abcam, ab290); guinea pig anti-vGlut1 antibody (Synaptic Systems, 135304); rabbit anti-calbindin antibody (Swant, CB38); rabbit anti-tyrosine hydroxylase antibody (Abcam, ab6211); rabbit anti-GAD2 antibody (Synaptic Systems, 198103); rabbit anti-GAD1/GAD2 antibody (Abcam ab11070); goat anti-parvalbumin antibody (Santa Cruz, sc-7449); rabbit anti-parvalbumin antibody (Abcam, ab11427); rabbit anti-metabotropic glutamate receptor antibody (Abcam, ab6438); rat anti-MBP antibody (Millipore, Mab386); rabbit anti-OMP antibody (Santa Cruz, sc-67219); rabbit anti-NG2 antibody (Millipore, AB5320); rabbit anti-CD11b (Novus Biologicals, NB110-89474); rabbit anti-S100b (Abcam, ab41548); rabbit anti-GFAP (Encor, RPCA-GFAP), and rabbit anti-NeuN (Millipore, MABN140).

Techniques: Binding Assay

The observation of MPS in neurons across different animal species. (A) Representative STORM image of immunolabeled βII spectrin in the axon of a cultured chicken neuron (Left) and the autocorrelation function calculated from the boxed region in this image (Right). (Scale bar: 2 µm.) (B) Representative STORM image of immunolabeled βII spectrin in the axon of a human iPS-derived motor neuron (Left) and the autocorrelation function calculated from this image (Right). (Scale bar: 2 µm.) (C) Two representative SIM images of β spectrin (UNC-70)-GFP in C. elegans neurons imaged directly in the animals (Left), and the corresponding autocorrelation functions of the boxed regions (Right). (Scale bar: 1 µm.) (D) Two representative SIM images of β spectrin-mMaple3 in Drosophila neurons imaged in Drosophila brain tissues (Left) and the corresponding autocorrelation functions of the boxed regions (Right). (Scale bar: 1 µm.)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Prevalent presence of periodic actin–spectrin-based membrane skeleton in a broad range of neuronal cell types and animal species

doi: 10.1073/pnas.1605707113

Figure Lengend Snippet: The observation of MPS in neurons across different animal species. (A) Representative STORM image of immunolabeled βII spectrin in the axon of a cultured chicken neuron (Left) and the autocorrelation function calculated from the boxed region in this image (Right). (Scale bar: 2 µm.) (B) Representative STORM image of immunolabeled βII spectrin in the axon of a human iPS-derived motor neuron (Left) and the autocorrelation function calculated from this image (Right). (Scale bar: 2 µm.) (C) Two representative SIM images of β spectrin (UNC-70)-GFP in C. elegans neurons imaged directly in the animals (Left), and the corresponding autocorrelation functions of the boxed regions (Right). (Scale bar: 1 µm.) (D) Two representative SIM images of β spectrin-mMaple3 in Drosophila neurons imaged in Drosophila brain tissues (Left) and the corresponding autocorrelation functions of the boxed regions (Right). (Scale bar: 1 µm.)

Article Snippet: The following primary antibodies were used in this study: guinea pig anti-MAP2 antibody (Synaptic Systems, 188002); mouse anti-βII spectrin antibody (BD Biosciences, 612563); rabbit anti-GFP antibody (Abcam, ab290); guinea pig anti-vGlut1 antibody (Synaptic Systems, 135304); rabbit anti-calbindin antibody (Swant, CB38); rabbit anti-tyrosine hydroxylase antibody (Abcam, ab6211); rabbit anti-GAD2 antibody (Synaptic Systems, 198103); rabbit anti-GAD1/GAD2 antibody (Abcam ab11070); goat anti-parvalbumin antibody (Santa Cruz, sc-7449); rabbit anti-parvalbumin antibody (Abcam, ab11427); rabbit anti-metabotropic glutamate receptor antibody (Abcam, ab6438); rat anti-MBP antibody (Millipore, Mab386); rabbit anti-OMP antibody (Santa Cruz, sc-67219); rabbit anti-NG2 antibody (Millipore, AB5320); rabbit anti-CD11b (Novus Biologicals, NB110-89474); rabbit anti-S100b (Abcam, ab41548); rabbit anti-GFAP (Encor, RPCA-GFAP), and rabbit anti-NeuN (Millipore, MABN140).

Techniques: Immunolabeling, Cell Culture, Derivative Assay