mouse anti synaptophysin Search Results


synaptophysin  (Bio-Rad)
92
Bio-Rad synaptophysin
FIGURE 1. (A–C) Representative sections illustrate immunohistochemical staining for <t>synaptophysin</t> (Syp) in the cerebral cortex and hippocampus (A), cerebellum (B), and corpus callosum (C) of untreated control mice. (D) Accumulation of synaptophysin- positive vesicles ¼ spheroids/ovoids/bulbs in the corpus callosum of a cuprizone-treated mouse. Arrows indicate the different sizes of synaptophysin-positive-spheroids; small [S],<2 mm (as in panel C); middle-sized [M], 2–7.5 mm; large [L], 8–15 mm, pathological size. (E, F) Graphs show numbers of different sized synaptophysin-positive spheroids in the corpus callosum during acute (5 weeks, E) and chronic (12 weeks, F) demyelination followed by remyelination after stopping cuprizone. Significant effects between different time points were calculated separately for each type/size of bulbs (*p <0.05, **p < 0.01, **p < 0.001). Significant effects for different cuprizone time treatments in comparison to the untreated control: #p< 0.05, ##p < 0.01, ###p < 0.001), n ¼ 4–6. (G, H) Representative images show synaptophysin-positive bulbs in the demyelinated hippocampus (G) and internal capsule (H). (I-K) Representative images show that synaptophysin does not colocalize with the oligodendroglial marker Nogo-A (I), the astrocytic marker GFAP (J), or with the microglial marker Iba-1 (K). Scale bar in (K)¼ 100 mm, applies to (I) and (J).
Synaptophysin, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mca1307  (Bio-Rad)
93
Bio-Rad mca1307
Antibodies used for the evaluation of injected pRPC and effect on host tissues
Mca1307, supplied by Bio-Rad, 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 mab igg1  (Valiant Co Ltd)
93
Valiant Co Ltd mouse mab igg1
Characterization of WR/Fas-SM( − ) and WR/Fas-SMS1 cells. (A) SM synthase activity of WR/Fas-SM(−) (lane 1) and WR/Fas-SMS1 (lane 2) cells. The cellular lipids were labeled with [ 14 C]serine, extracted by the Bligh and Dyer method (reference ), and assessed by TLC. PE, phosphatidylethanolamine; PS, phosphatidylserine. (B) Analysis of membrane SM expression by confocal microscopy. Cells were stained with lysenin-MBP, and FITC-conjugated anti–mouse <t>IgG</t> mAb, then examined by laser scan confocal microscopy. (C) FACS analysis of membrane sphingolipids. To detect membrane SM, cells were stained with lysenin-MBP (Lysenin), CH11, and FITC-conjugated anti–mouse IgG mAb. Surface expression of ganglioside GM1, cholesterol, and human Fas were analyzed using FITC-conjugated CTx, cholesterol-PEG (cholesterol), or anti-Fas mAB (h-Fas), respectively.
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mouse monoclonal anti-synaptophysin antibody  (Becton Dickinson)
90
Becton Dickinson mouse monoclonal anti-synaptophysin antibody
PACAP significantly increased the number of function synapse in the primary cultured hippocampal neurons. A, Representative fluorescence images of the primary hippocampal neuronal dendrites at DIV21. The cells were fixed at DIV21 and immunostained using <t>anti-synaptophysin</t> (magenta) and anti-PSD95 (green) antibodies. Magnification of the area marked with a white dot box. Scale bar, 10 μm. B, Primary hippocampal neurons were treated for 2 d with 1, 10, or 100 nm PACAP. Scale bar, 10 μm. C, Quantification of the number of PSD-95-positive spines that were apposed to synaptophysin-positive presynaptic terminals. Values indicate mean ± SEM on 60 neurons from three independent experiments. **p < 0.01 versus vehicle (one-way ANOVA followed by Tukey–Kramer test). D, Immunoblot of cell surface biotinylated α1 sodium potassium ATPase, NR1, and NR2B from the primary cultured hippocampal neurons at DIV14. Surface biotinylation assay was performed after 2 d treatment with vehicle or 10 nm PACAP. E, Quantification of the cell surface expression levels of NR1 and NR2B were normalized to the levels of α1 sodium potassium ATPase. Values indicate mean ± SEM from five independent experiments. *p < 0.05 versus vehicle (Student's t test). F, Representative immunoblots showing expression of pERK1/2 and ERK2. Synaptic NMDAR activity in the primary cultured hippocampal neurons at DIV14 were treated with vehicle or 500 μm 4-AP and 50 μm bicuculline (bic) for 5 or 30 min after 2 d treatment with vehicle or 10 nm PACAP. 4-AP is a potassium channel blocker, and bicuculline is a GABAA receptor blocker. G, Quantification of the expression levels of pERK2 were normalized to the levels of ERK2. Values indicate mean ± SEM from four independent experiments. *p < 0.05 versus vehicle or 0 min (two-way ANOVA followed by Tukey–Kramer test).
Mouse Monoclonal Anti Synaptophysin 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
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anti‑synaptophysin mouse monoclonal anti‑syn ab clone sy38  (Merck KGaA)
90
Merck KGaA anti‑synaptophysin mouse monoclonal anti‑syn ab clone sy38
PACAP significantly increased the number of function synapse in the primary cultured hippocampal neurons. A, Representative fluorescence images of the primary hippocampal neuronal dendrites at DIV21. The cells were fixed at DIV21 and immunostained using <t>anti-synaptophysin</t> (magenta) and anti-PSD95 (green) antibodies. Magnification of the area marked with a white dot box. Scale bar, 10 μm. B, Primary hippocampal neurons were treated for 2 d with 1, 10, or 100 nm PACAP. Scale bar, 10 μm. C, Quantification of the number of PSD-95-positive spines that were apposed to synaptophysin-positive presynaptic terminals. Values indicate mean ± SEM on 60 neurons from three independent experiments. **p < 0.01 versus vehicle (one-way ANOVA followed by Tukey–Kramer test). D, Immunoblot of cell surface biotinylated α1 sodium potassium ATPase, NR1, and NR2B from the primary cultured hippocampal neurons at DIV14. Surface biotinylation assay was performed after 2 d treatment with vehicle or 10 nm PACAP. E, Quantification of the cell surface expression levels of NR1 and NR2B were normalized to the levels of α1 sodium potassium ATPase. Values indicate mean ± SEM from five independent experiments. *p < 0.05 versus vehicle (Student's t test). F, Representative immunoblots showing expression of pERK1/2 and ERK2. Synaptic NMDAR activity in the primary cultured hippocampal neurons at DIV14 were treated with vehicle or 500 μm 4-AP and 50 μm bicuculline (bic) for 5 or 30 min after 2 d treatment with vehicle or 10 nm PACAP. 4-AP is a potassium channel blocker, and bicuculline is a GABAA receptor blocker. G, Quantification of the expression levels of pERK2 were normalized to the levels of ERK2. Values indicate mean ± SEM from four independent experiments. *p < 0.05 versus vehicle or 0 min (two-way ANOVA followed by Tukey–Kramer test).
Anti‑Synaptophysin Mouse Monoclonal Anti‑Syn Ab Clone Sy38, supplied by Merck KGaA, 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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mouse anti-synaptophysin genetex gtx38985  (GeneTex)
90
GeneTex mouse anti-synaptophysin genetex gtx38985
PACAP significantly increased the number of function synapse in the primary cultured hippocampal neurons. A, Representative fluorescence images of the primary hippocampal neuronal dendrites at DIV21. The cells were fixed at DIV21 and immunostained using <t>anti-synaptophysin</t> (magenta) and anti-PSD95 (green) antibodies. Magnification of the area marked with a white dot box. Scale bar, 10 μm. B, Primary hippocampal neurons were treated for 2 d with 1, 10, or 100 nm PACAP. Scale bar, 10 μm. C, Quantification of the number of PSD-95-positive spines that were apposed to synaptophysin-positive presynaptic terminals. Values indicate mean ± SEM on 60 neurons from three independent experiments. **p < 0.01 versus vehicle (one-way ANOVA followed by Tukey–Kramer test). D, Immunoblot of cell surface biotinylated α1 sodium potassium ATPase, NR1, and NR2B from the primary cultured hippocampal neurons at DIV14. Surface biotinylation assay was performed after 2 d treatment with vehicle or 10 nm PACAP. E, Quantification of the cell surface expression levels of NR1 and NR2B were normalized to the levels of α1 sodium potassium ATPase. Values indicate mean ± SEM from five independent experiments. *p < 0.05 versus vehicle (Student's t test). F, Representative immunoblots showing expression of pERK1/2 and ERK2. Synaptic NMDAR activity in the primary cultured hippocampal neurons at DIV14 were treated with vehicle or 500 μm 4-AP and 50 μm bicuculline (bic) for 5 or 30 min after 2 d treatment with vehicle or 10 nm PACAP. 4-AP is a potassium channel blocker, and bicuculline is a GABAA receptor blocker. G, Quantification of the expression levels of pERK2 were normalized to the levels of ERK2. Values indicate mean ± SEM from four independent experiments. *p < 0.05 versus vehicle or 0 min (two-way ANOVA followed by Tukey–Kramer test).
Mouse Anti Synaptophysin Genetex Gtx38985, supplied by GeneTex, 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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mouse anti-synaptophysin  (Becton Dickinson)
90
Becton Dickinson mouse anti-synaptophysin
PACAP significantly increased the number of function synapse in the primary cultured hippocampal neurons. A, Representative fluorescence images of the primary hippocampal neuronal dendrites at DIV21. The cells were fixed at DIV21 and immunostained using <t>anti-synaptophysin</t> (magenta) and anti-PSD95 (green) antibodies. Magnification of the area marked with a white dot box. Scale bar, 10 μm. B, Primary hippocampal neurons were treated for 2 d with 1, 10, or 100 nm PACAP. Scale bar, 10 μm. C, Quantification of the number of PSD-95-positive spines that were apposed to synaptophysin-positive presynaptic terminals. Values indicate mean ± SEM on 60 neurons from three independent experiments. **p < 0.01 versus vehicle (one-way ANOVA followed by Tukey–Kramer test). D, Immunoblot of cell surface biotinylated α1 sodium potassium ATPase, NR1, and NR2B from the primary cultured hippocampal neurons at DIV14. Surface biotinylation assay was performed after 2 d treatment with vehicle or 10 nm PACAP. E, Quantification of the cell surface expression levels of NR1 and NR2B were normalized to the levels of α1 sodium potassium ATPase. Values indicate mean ± SEM from five independent experiments. *p < 0.05 versus vehicle (Student's t test). F, Representative immunoblots showing expression of pERK1/2 and ERK2. Synaptic NMDAR activity in the primary cultured hippocampal neurons at DIV14 were treated with vehicle or 500 μm 4-AP and 50 μm bicuculline (bic) for 5 or 30 min after 2 d treatment with vehicle or 10 nm PACAP. 4-AP is a potassium channel blocker, and bicuculline is a GABAA receptor blocker. G, Quantification of the expression levels of pERK2 were normalized to the levels of ERK2. Values indicate mean ± SEM from four independent experiments. *p < 0.05 versus vehicle or 0 min (two-way ANOVA followed by Tukey–Kramer test).
Mouse Anti Synaptophysin, 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
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mouse anti-synaptophysin protein monoclonal antibody  (Becton Dickinson)
90
Becton Dickinson mouse anti-synaptophysin protein monoclonal antibody
PACAP significantly increased the number of function synapse in the primary cultured hippocampal neurons. A, Representative fluorescence images of the primary hippocampal neuronal dendrites at DIV21. The cells were fixed at DIV21 and immunostained using <t>anti-synaptophysin</t> (magenta) and anti-PSD95 (green) antibodies. Magnification of the area marked with a white dot box. Scale bar, 10 μm. B, Primary hippocampal neurons were treated for 2 d with 1, 10, or 100 nm PACAP. Scale bar, 10 μm. C, Quantification of the number of PSD-95-positive spines that were apposed to synaptophysin-positive presynaptic terminals. Values indicate mean ± SEM on 60 neurons from three independent experiments. **p < 0.01 versus vehicle (one-way ANOVA followed by Tukey–Kramer test). D, Immunoblot of cell surface biotinylated α1 sodium potassium ATPase, NR1, and NR2B from the primary cultured hippocampal neurons at DIV14. Surface biotinylation assay was performed after 2 d treatment with vehicle or 10 nm PACAP. E, Quantification of the cell surface expression levels of NR1 and NR2B were normalized to the levels of α1 sodium potassium ATPase. Values indicate mean ± SEM from five independent experiments. *p < 0.05 versus vehicle (Student's t test). F, Representative immunoblots showing expression of pERK1/2 and ERK2. Synaptic NMDAR activity in the primary cultured hippocampal neurons at DIV14 were treated with vehicle or 500 μm 4-AP and 50 μm bicuculline (bic) for 5 or 30 min after 2 d treatment with vehicle or 10 nm PACAP. 4-AP is a potassium channel blocker, and bicuculline is a GABAA receptor blocker. G, Quantification of the expression levels of pERK2 were normalized to the levels of ERK2. Values indicate mean ± SEM from four independent experiments. *p < 0.05 versus vehicle or 0 min (two-way ANOVA followed by Tukey–Kramer test).
Mouse Anti Synaptophysin Protein Monoclonal 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
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mouse anti-synaptophysin  (Enzo Biochem)
90
Enzo Biochem mouse anti-synaptophysin
PACAP significantly increased the number of function synapse in the primary cultured hippocampal neurons. A, Representative fluorescence images of the primary hippocampal neuronal dendrites at DIV21. The cells were fixed at DIV21 and immunostained using <t>anti-synaptophysin</t> (magenta) and anti-PSD95 (green) antibodies. Magnification of the area marked with a white dot box. Scale bar, 10 μm. B, Primary hippocampal neurons were treated for 2 d with 1, 10, or 100 nm PACAP. Scale bar, 10 μm. C, Quantification of the number of PSD-95-positive spines that were apposed to synaptophysin-positive presynaptic terminals. Values indicate mean ± SEM on 60 neurons from three independent experiments. **p < 0.01 versus vehicle (one-way ANOVA followed by Tukey–Kramer test). D, Immunoblot of cell surface biotinylated α1 sodium potassium ATPase, NR1, and NR2B from the primary cultured hippocampal neurons at DIV14. Surface biotinylation assay was performed after 2 d treatment with vehicle or 10 nm PACAP. E, Quantification of the cell surface expression levels of NR1 and NR2B were normalized to the levels of α1 sodium potassium ATPase. Values indicate mean ± SEM from five independent experiments. *p < 0.05 versus vehicle (Student's t test). F, Representative immunoblots showing expression of pERK1/2 and ERK2. Synaptic NMDAR activity in the primary cultured hippocampal neurons at DIV14 were treated with vehicle or 500 μm 4-AP and 50 μm bicuculline (bic) for 5 or 30 min after 2 d treatment with vehicle or 10 nm PACAP. 4-AP is a potassium channel blocker, and bicuculline is a GABAA receptor blocker. G, Quantification of the expression levels of pERK2 were normalized to the levels of ERK2. Values indicate mean ± SEM from four independent experiments. *p < 0.05 versus vehicle or 0 min (two-way ANOVA followed by Tukey–Kramer test).
Mouse Anti Synaptophysin, supplied by Enzo Biochem, 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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mouse anti-synaptophysin mab329/ab _ 95786  (Merck KGaA)
90
Merck KGaA mouse anti-synaptophysin mab329/ab _ 95786
PACAP significantly increased the number of function synapse in the primary cultured hippocampal neurons. A, Representative fluorescence images of the primary hippocampal neuronal dendrites at DIV21. The cells were fixed at DIV21 and immunostained using <t>anti-synaptophysin</t> (magenta) and anti-PSD95 (green) antibodies. Magnification of the area marked with a white dot box. Scale bar, 10 μm. B, Primary hippocampal neurons were treated for 2 d with 1, 10, or 100 nm PACAP. Scale bar, 10 μm. C, Quantification of the number of PSD-95-positive spines that were apposed to synaptophysin-positive presynaptic terminals. Values indicate mean ± SEM on 60 neurons from three independent experiments. **p < 0.01 versus vehicle (one-way ANOVA followed by Tukey–Kramer test). D, Immunoblot of cell surface biotinylated α1 sodium potassium ATPase, NR1, and NR2B from the primary cultured hippocampal neurons at DIV14. Surface biotinylation assay was performed after 2 d treatment with vehicle or 10 nm PACAP. E, Quantification of the cell surface expression levels of NR1 and NR2B were normalized to the levels of α1 sodium potassium ATPase. Values indicate mean ± SEM from five independent experiments. *p < 0.05 versus vehicle (Student's t test). F, Representative immunoblots showing expression of pERK1/2 and ERK2. Synaptic NMDAR activity in the primary cultured hippocampal neurons at DIV14 were treated with vehicle or 500 μm 4-AP and 50 μm bicuculline (bic) for 5 or 30 min after 2 d treatment with vehicle or 10 nm PACAP. 4-AP is a potassium channel blocker, and bicuculline is a GABAA receptor blocker. G, Quantification of the expression levels of pERK2 were normalized to the levels of ERK2. Values indicate mean ± SEM from four independent experiments. *p < 0.05 versus vehicle or 0 min (two-way ANOVA followed by Tukey–Kramer test).
Mouse Anti Synaptophysin Mab329/Ab 95786, supplied by Merck KGaA, 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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anti-synaptophysin mouse monoclonal  (Becton Dickinson)
90
Becton Dickinson anti-synaptophysin mouse monoclonal
PACAP significantly increased the number of function synapse in the primary cultured hippocampal neurons. A, Representative fluorescence images of the primary hippocampal neuronal dendrites at DIV21. The cells were fixed at DIV21 and immunostained using <t>anti-synaptophysin</t> (magenta) and anti-PSD95 (green) antibodies. Magnification of the area marked with a white dot box. Scale bar, 10 μm. B, Primary hippocampal neurons were treated for 2 d with 1, 10, or 100 nm PACAP. Scale bar, 10 μm. C, Quantification of the number of PSD-95-positive spines that were apposed to synaptophysin-positive presynaptic terminals. Values indicate mean ± SEM on 60 neurons from three independent experiments. **p < 0.01 versus vehicle (one-way ANOVA followed by Tukey–Kramer test). D, Immunoblot of cell surface biotinylated α1 sodium potassium ATPase, NR1, and NR2B from the primary cultured hippocampal neurons at DIV14. Surface biotinylation assay was performed after 2 d treatment with vehicle or 10 nm PACAP. E, Quantification of the cell surface expression levels of NR1 and NR2B were normalized to the levels of α1 sodium potassium ATPase. Values indicate mean ± SEM from five independent experiments. *p < 0.05 versus vehicle (Student's t test). F, Representative immunoblots showing expression of pERK1/2 and ERK2. Synaptic NMDAR activity in the primary cultured hippocampal neurons at DIV14 were treated with vehicle or 500 μm 4-AP and 50 μm bicuculline (bic) for 5 or 30 min after 2 d treatment with vehicle or 10 nm PACAP. 4-AP is a potassium channel blocker, and bicuculline is a GABAA receptor blocker. G, Quantification of the expression levels of pERK2 were normalized to the levels of ERK2. Values indicate mean ± SEM from four independent experiments. *p < 0.05 versus vehicle or 0 min (two-way ANOVA followed by Tukey–Kramer test).
Anti Synaptophysin Mouse Monoclonal, 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
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mouse anti-synaptophysin, 1 : 100, assaydesigns  (Assay Designs Inc)
90
Assay Designs Inc mouse anti-synaptophysin, 1 : 100, assaydesigns
PACAP significantly increased the number of function synapse in the primary cultured hippocampal neurons. A, Representative fluorescence images of the primary hippocampal neuronal dendrites at DIV21. The cells were fixed at DIV21 and immunostained using <t>anti-synaptophysin</t> (magenta) and anti-PSD95 (green) antibodies. Magnification of the area marked with a white dot box. Scale bar, 10 μm. B, Primary hippocampal neurons were treated for 2 d with 1, 10, or 100 nm PACAP. Scale bar, 10 μm. C, Quantification of the number of PSD-95-positive spines that were apposed to synaptophysin-positive presynaptic terminals. Values indicate mean ± SEM on 60 neurons from three independent experiments. **p < 0.01 versus vehicle (one-way ANOVA followed by Tukey–Kramer test). D, Immunoblot of cell surface biotinylated α1 sodium potassium ATPase, NR1, and NR2B from the primary cultured hippocampal neurons at DIV14. Surface biotinylation assay was performed after 2 d treatment with vehicle or 10 nm PACAP. E, Quantification of the cell surface expression levels of NR1 and NR2B were normalized to the levels of α1 sodium potassium ATPase. Values indicate mean ± SEM from five independent experiments. *p < 0.05 versus vehicle (Student's t test). F, Representative immunoblots showing expression of pERK1/2 and ERK2. Synaptic NMDAR activity in the primary cultured hippocampal neurons at DIV14 were treated with vehicle or 500 μm 4-AP and 50 μm bicuculline (bic) for 5 or 30 min after 2 d treatment with vehicle or 10 nm PACAP. 4-AP is a potassium channel blocker, and bicuculline is a GABAA receptor blocker. G, Quantification of the expression levels of pERK2 were normalized to the levels of ERK2. Values indicate mean ± SEM from four independent experiments. *p < 0.05 versus vehicle or 0 min (two-way ANOVA followed by Tukey–Kramer test).
Mouse Anti Synaptophysin, 1 : 100, Assaydesigns, supplied by Assay Designs 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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Image Search Results


FIGURE 1. (A–C) Representative sections illustrate immunohistochemical staining for synaptophysin (Syp) in the cerebral cortex and hippocampus (A), cerebellum (B), and corpus callosum (C) of untreated control mice. (D) Accumulation of synaptophysin- positive vesicles ¼ spheroids/ovoids/bulbs in the corpus callosum of a cuprizone-treated mouse. Arrows indicate the different sizes of synaptophysin-positive-spheroids; small [S],<2 mm (as in panel C); middle-sized [M], 2–7.5 mm; large [L], 8–15 mm, pathological size. (E, F) Graphs show numbers of different sized synaptophysin-positive spheroids in the corpus callosum during acute (5 weeks, E) and chronic (12 weeks, F) demyelination followed by remyelination after stopping cuprizone. Significant effects between different time points were calculated separately for each type/size of bulbs (*p <0.05, **p < 0.01, **p < 0.001). Significant effects for different cuprizone time treatments in comparison to the untreated control: #p< 0.05, ##p < 0.01, ###p < 0.001), n ¼ 4–6. (G, H) Representative images show synaptophysin-positive bulbs in the demyelinated hippocampus (G) and internal capsule (H). (I-K) Representative images show that synaptophysin does not colocalize with the oligodendroglial marker Nogo-A (I), the astrocytic marker GFAP (J), or with the microglial marker Iba-1 (K). Scale bar in (K)¼ 100 mm, applies to (I) and (J).

Journal: Journal of neuropathology and experimental neurology

Article Title: Synaptophysin Is a Reliable Marker for Axonal Damage.

doi: 10.1093/jnen/nlw114

Figure Lengend Snippet: FIGURE 1. (A–C) Representative sections illustrate immunohistochemical staining for synaptophysin (Syp) in the cerebral cortex and hippocampus (A), cerebellum (B), and corpus callosum (C) of untreated control mice. (D) Accumulation of synaptophysin- positive vesicles ¼ spheroids/ovoids/bulbs in the corpus callosum of a cuprizone-treated mouse. Arrows indicate the different sizes of synaptophysin-positive-spheroids; small [S],<2 mm (as in panel C); middle-sized [M], 2–7.5 mm; large [L], 8–15 mm, pathological size. (E, F) Graphs show numbers of different sized synaptophysin-positive spheroids in the corpus callosum during acute (5 weeks, E) and chronic (12 weeks, F) demyelination followed by remyelination after stopping cuprizone. Significant effects between different time points were calculated separately for each type/size of bulbs (*p <0.05, **p < 0.01, **p < 0.001). Significant effects for different cuprizone time treatments in comparison to the untreated control: #p< 0.05, ##p < 0.01, ###p < 0.001), n ¼ 4–6. (G, H) Representative images show synaptophysin-positive bulbs in the demyelinated hippocampus (G) and internal capsule (H). (I-K) Representative images show that synaptophysin does not colocalize with the oligodendroglial marker Nogo-A (I), the astrocytic marker GFAP (J), or with the microglial marker Iba-1 (K). Scale bar in (K)¼ 100 mm, applies to (I) and (J).

Article Snippet: We used 2 different antibodies for synaptophysin: a mouse monoclonal IgG from Serotec and rabbit monoclonal from Abcam.

Techniques: Immunohistochemical staining, Staining, Control, Comparison, Marker

FIGURE 2. (A, B) Graphs represent the quantitative analysis of APP, synaptophysin (Syp), and double-positive spheroids in the corpus callosum of cuprizone-treated mice with acute demyelination (A) and chronic demyelination (B). Two-way ANOVA analysis identify that there were no significant differences between numbers of APP- and synaptophysin-positive spheroids during

Journal: Journal of neuropathology and experimental neurology

Article Title: Synaptophysin Is a Reliable Marker for Axonal Damage.

doi: 10.1093/jnen/nlw114

Figure Lengend Snippet: FIGURE 2. (A, B) Graphs represent the quantitative analysis of APP, synaptophysin (Syp), and double-positive spheroids in the corpus callosum of cuprizone-treated mice with acute demyelination (A) and chronic demyelination (B). Two-way ANOVA analysis identify that there were no significant differences between numbers of APP- and synaptophysin-positive spheroids during

Article Snippet: We used 2 different antibodies for synaptophysin: a mouse monoclonal IgG from Serotec and rabbit monoclonal from Abcam.

Techniques:

FIGURE 3. Immunohistochemical staining of the corpus callosum evaluating phosphorylation of neurofilaments and their colocalization with synaptophysin (Syp) during cuprizone-induced demyelination. (A–H) SMI-32 detecting non-phosphorylated heavy neurofilaments are shown in panels (A, C, E, G). SMI-312 detecting phosphorylated neurofilaments H and M are

Journal: Journal of neuropathology and experimental neurology

Article Title: Synaptophysin Is a Reliable Marker for Axonal Damage.

doi: 10.1093/jnen/nlw114

Figure Lengend Snippet: FIGURE 3. Immunohistochemical staining of the corpus callosum evaluating phosphorylation of neurofilaments and their colocalization with synaptophysin (Syp) during cuprizone-induced demyelination. (A–H) SMI-32 detecting non-phosphorylated heavy neurofilaments are shown in panels (A, C, E, G). SMI-312 detecting phosphorylated neurofilaments H and M are

Article Snippet: We used 2 different antibodies for synaptophysin: a mouse monoclonal IgG from Serotec and rabbit monoclonal from Abcam.

Techniques: Immunohistochemical staining, Staining, Phospho-proteomics

FIGURE 5. (A-P) Representative sections show immunostaining in the corpus callosum for synaptophysin/Iba-1 (A-F) and synaptophysin/ PLP (G–L) during acute and chronic cuprizone induced demyelination and subsequent remyelination. Inserts in (A–F) show the morphology/shape of microglia identifying their activation state. Inserts in (G–L) show higher magnification of synaptophysin accumulations. Panels (M) and (N) are confocal images from RCA-1 (microglia activation marker, green) and synaptophysin (red) during inflammation in cuprizone-treated mice. Panels (O) and (P) show remyelinated axons (PLP, green) with internal synaptophysin accumulation (red).

Journal: Journal of neuropathology and experimental neurology

Article Title: Synaptophysin Is a Reliable Marker for Axonal Damage.

doi: 10.1093/jnen/nlw114

Figure Lengend Snippet: FIGURE 5. (A-P) Representative sections show immunostaining in the corpus callosum for synaptophysin/Iba-1 (A-F) and synaptophysin/ PLP (G–L) during acute and chronic cuprizone induced demyelination and subsequent remyelination. Inserts in (A–F) show the morphology/shape of microglia identifying their activation state. Inserts in (G–L) show higher magnification of synaptophysin accumulations. Panels (M) and (N) are confocal images from RCA-1 (microglia activation marker, green) and synaptophysin (red) during inflammation in cuprizone-treated mice. Panels (O) and (P) show remyelinated axons (PLP, green) with internal synaptophysin accumulation (red).

Article Snippet: We used 2 different antibodies for synaptophysin: a mouse monoclonal IgG from Serotec and rabbit monoclonal from Abcam.

Techniques: Immunostaining, Activation Assay, Marker

FIGURE 6. The appearance of synaptophysin (Syp)-positive pathological (medium-and large-sized) spheroids was compared and correlated with the extent of demyelination (proteolipid protein: PLP, myelin basic protein: MBP) and microglia infiltration (Iba-1) induced by cuprizone feeding (acute or chronic). (A, B) Graphs show the extent of microglia accumulation, appearance of pathological synaptophysin bulbs and the extent of de- and remyelination as judged by scoring of MBP and PLP immunostaining. Score of 3 represents complete myelination; score of 0 represents complete demyelination. (C–H) Correlation analyses of synaptophysin-positive bulbs with the extent of de- and remyelination and microglia numbers (n ¼ 5–6).

Journal: Journal of neuropathology and experimental neurology

Article Title: Synaptophysin Is a Reliable Marker for Axonal Damage.

doi: 10.1093/jnen/nlw114

Figure Lengend Snippet: FIGURE 6. The appearance of synaptophysin (Syp)-positive pathological (medium-and large-sized) spheroids was compared and correlated with the extent of demyelination (proteolipid protein: PLP, myelin basic protein: MBP) and microglia infiltration (Iba-1) induced by cuprizone feeding (acute or chronic). (A, B) Graphs show the extent of microglia accumulation, appearance of pathological synaptophysin bulbs and the extent of de- and remyelination as judged by scoring of MBP and PLP immunostaining. Score of 3 represents complete myelination; score of 0 represents complete demyelination. (C–H) Correlation analyses of synaptophysin-positive bulbs with the extent of de- and remyelination and microglia numbers (n ¼ 5–6).

Article Snippet: We used 2 different antibodies for synaptophysin: a mouse monoclonal IgG from Serotec and rabbit monoclonal from Abcam.

Techniques: Immunostaining

FIGURE 8. (A–C) Graphs show microglia/macrophage infiltration (A) and the course of demyelination (B) during Theiler’s virus induced encephalomyelitis in the thoracic segment of murine spinal cord. (C) Panel shows the appearance of pathologic accumulation of synaptophysin (Syp). Significant effects between different time points were indicated as: *p < 0.05, **p < 0.01, **p < 0.001. Significant effects for different time treatments in comparison to the untreated control are indicated as: # p < 0.05, ## p < 0.01, ### p < 0.001), n ¼ 4–6. (D–H) Representative images of thoracic spinal cord segment illustrate infiltration of microglia/macrophage at different days after the injection of Theiler’s murine encephalomyelitis virus. (I–U) Panel (I) shows the appearance of synaptophysin-positive bulbs exclusively in the demyelinated area; (J) and (K) illustrate the appearance of synaptophysin-positive spheroids surrounded by activated microglia/macrophages. (L–N) Colocalization of synaptophysin and APP in the spheroid structures. (O–U) Serial staining of the same spinal cord area during Theiler’s virus induced encephalomyelitis (O, synaptophysin/PLP double staining; P, synaptophysin/Iba-1 double staining; R, synaptophysin/APP double staining; S–U, higher magnification of area shown in R).

Journal: Journal of neuropathology and experimental neurology

Article Title: Synaptophysin Is a Reliable Marker for Axonal Damage.

doi: 10.1093/jnen/nlw114

Figure Lengend Snippet: FIGURE 8. (A–C) Graphs show microglia/macrophage infiltration (A) and the course of demyelination (B) during Theiler’s virus induced encephalomyelitis in the thoracic segment of murine spinal cord. (C) Panel shows the appearance of pathologic accumulation of synaptophysin (Syp). Significant effects between different time points were indicated as: *p < 0.05, **p < 0.01, **p < 0.001. Significant effects for different time treatments in comparison to the untreated control are indicated as: # p < 0.05, ## p < 0.01, ### p < 0.001), n ¼ 4–6. (D–H) Representative images of thoracic spinal cord segment illustrate infiltration of microglia/macrophage at different days after the injection of Theiler’s murine encephalomyelitis virus. (I–U) Panel (I) shows the appearance of synaptophysin-positive bulbs exclusively in the demyelinated area; (J) and (K) illustrate the appearance of synaptophysin-positive spheroids surrounded by activated microglia/macrophages. (L–N) Colocalization of synaptophysin and APP in the spheroid structures. (O–U) Serial staining of the same spinal cord area during Theiler’s virus induced encephalomyelitis (O, synaptophysin/PLP double staining; P, synaptophysin/Iba-1 double staining; R, synaptophysin/APP double staining; S–U, higher magnification of area shown in R).

Article Snippet: We used 2 different antibodies for synaptophysin: a mouse monoclonal IgG from Serotec and rabbit monoclonal from Abcam.

Techniques: Virus, Comparison, Control, Injection, Staining, Double Staining

FIGURE 9. Representative images show the expression of synaptophysin (Syp) in CNS tissue of 2 different patients with MS. (A– H) Different immunohistochemical stainings from a chronic active MS lesion (serial sections). Panels (E) and (F) show the higher magnifications from boxed areas 1 in (C) and (D). Panels (G) and (H) demonstrate higher magnification from boxed areas 2 in (C) and D. (I–K) Representative double immunostaining for synaptophysin and Iba-1 demonstrate microglia accumulation (I) and proving colocalization of APP and synaptophysin (J, K). (L–O) Demyelination/inflammation/axonal damage in another MS lesion. DAB staining in (L) and (M) illustrate the extent of demyelination (PLP) and inflammation (LN3 marks HLA-DR-positive cells: lymphocytes and macrophages) in this MS lesion. Panels N and O show higher magnification of the boxed are from this lesion in (L) and (M) demonstrating double expression of APP and synaptophysin in (N) and double staining of PLP and synaptophysin in (O). (P–U) Corpora amylacea are immunostain-positive for APP (P, S), and NeuN (U), but not synaptophysin (R, T).

Journal: Journal of neuropathology and experimental neurology

Article Title: Synaptophysin Is a Reliable Marker for Axonal Damage.

doi: 10.1093/jnen/nlw114

Figure Lengend Snippet: FIGURE 9. Representative images show the expression of synaptophysin (Syp) in CNS tissue of 2 different patients with MS. (A– H) Different immunohistochemical stainings from a chronic active MS lesion (serial sections). Panels (E) and (F) show the higher magnifications from boxed areas 1 in (C) and (D). Panels (G) and (H) demonstrate higher magnification from boxed areas 2 in (C) and D. (I–K) Representative double immunostaining for synaptophysin and Iba-1 demonstrate microglia accumulation (I) and proving colocalization of APP and synaptophysin (J, K). (L–O) Demyelination/inflammation/axonal damage in another MS lesion. DAB staining in (L) and (M) illustrate the extent of demyelination (PLP) and inflammation (LN3 marks HLA-DR-positive cells: lymphocytes and macrophages) in this MS lesion. Panels N and O show higher magnification of the boxed are from this lesion in (L) and (M) demonstrating double expression of APP and synaptophysin in (N) and double staining of PLP and synaptophysin in (O). (P–U) Corpora amylacea are immunostain-positive for APP (P, S), and NeuN (U), but not synaptophysin (R, T).

Article Snippet: We used 2 different antibodies for synaptophysin: a mouse monoclonal IgG from Serotec and rabbit monoclonal from Abcam.

Techniques: Expressing, Immunohistochemical staining, Double Immunostaining, Staining, Double Staining

Antibodies used for the evaluation of injected pRPC and effect on host tissues

Journal: Journal of Cellular and Molecular Medicine

Article Title: In vivo study to assess dosage of allogeneic pig retinal progenitor cells: Long‐term survival, engraftment, differentiation and safety

doi: 10.1111/jcmm.17332

Figure Lengend Snippet: Antibodies used for the evaluation of injected pRPC and effect on host tissues

Article Snippet: Synaptophysin , Mouse , Synaptophysin , Fiber synapses , Serotec , MCA1307 , 1:25.

Techniques: Injection

Characterization of WR/Fas-SM( − ) and WR/Fas-SMS1 cells. (A) SM synthase activity of WR/Fas-SM(−) (lane 1) and WR/Fas-SMS1 (lane 2) cells. The cellular lipids were labeled with [ 14 C]serine, extracted by the Bligh and Dyer method (reference ), and assessed by TLC. PE, phosphatidylethanolamine; PS, phosphatidylserine. (B) Analysis of membrane SM expression by confocal microscopy. Cells were stained with lysenin-MBP, and FITC-conjugated anti–mouse IgG mAb, then examined by laser scan confocal microscopy. (C) FACS analysis of membrane sphingolipids. To detect membrane SM, cells were stained with lysenin-MBP (Lysenin), CH11, and FITC-conjugated anti–mouse IgG mAb. Surface expression of ganglioside GM1, cholesterol, and human Fas were analyzed using FITC-conjugated CTx, cholesterol-PEG (cholesterol), or anti-Fas mAB (h-Fas), respectively.

Journal: The Journal of Experimental Medicine

Article Title: Role of membrane sphingomyelin and ceramide in platform formation for Fas-mediated apoptosis

doi: 10.1084/jem.20041685

Figure Lengend Snippet: Characterization of WR/Fas-SM( − ) and WR/Fas-SMS1 cells. (A) SM synthase activity of WR/Fas-SM(−) (lane 1) and WR/Fas-SMS1 (lane 2) cells. The cellular lipids were labeled with [ 14 C]serine, extracted by the Bligh and Dyer method (reference ), and assessed by TLC. PE, phosphatidylethanolamine; PS, phosphatidylserine. (B) Analysis of membrane SM expression by confocal microscopy. Cells were stained with lysenin-MBP, and FITC-conjugated anti–mouse IgG mAb, then examined by laser scan confocal microscopy. (C) FACS analysis of membrane sphingolipids. To detect membrane SM, cells were stained with lysenin-MBP (Lysenin), CH11, and FITC-conjugated anti–mouse IgG mAb. Surface expression of ganglioside GM1, cholesterol, and human Fas were analyzed using FITC-conjugated CTx, cholesterol-PEG (cholesterol), or anti-Fas mAB (h-Fas), respectively.

Article Snippet: Aggregated Fas was detected by 3D5, a mouse mAb (IgG1) specific for the intracellular death domain of human Fas (Qbiogene).

Techniques: Activity Assay, Labeling, Expressing, Confocal Microscopy, Staining

PACAP significantly increased the number of function synapse in the primary cultured hippocampal neurons. A, Representative fluorescence images of the primary hippocampal neuronal dendrites at DIV21. The cells were fixed at DIV21 and immunostained using anti-synaptophysin (magenta) and anti-PSD95 (green) antibodies. Magnification of the area marked with a white dot box. Scale bar, 10 μm. B, Primary hippocampal neurons were treated for 2 d with 1, 10, or 100 nm PACAP. Scale bar, 10 μm. C, Quantification of the number of PSD-95-positive spines that were apposed to synaptophysin-positive presynaptic terminals. Values indicate mean ± SEM on 60 neurons from three independent experiments. **p < 0.01 versus vehicle (one-way ANOVA followed by Tukey–Kramer test). D, Immunoblot of cell surface biotinylated α1 sodium potassium ATPase, NR1, and NR2B from the primary cultured hippocampal neurons at DIV14. Surface biotinylation assay was performed after 2 d treatment with vehicle or 10 nm PACAP. E, Quantification of the cell surface expression levels of NR1 and NR2B were normalized to the levels of α1 sodium potassium ATPase. Values indicate mean ± SEM from five independent experiments. *p < 0.05 versus vehicle (Student's t test). F, Representative immunoblots showing expression of pERK1/2 and ERK2. Synaptic NMDAR activity in the primary cultured hippocampal neurons at DIV14 were treated with vehicle or 500 μm 4-AP and 50 μm bicuculline (bic) for 5 or 30 min after 2 d treatment with vehicle or 10 nm PACAP. 4-AP is a potassium channel blocker, and bicuculline is a GABAA receptor blocker. G, Quantification of the expression levels of pERK2 were normalized to the levels of ERK2. Values indicate mean ± SEM from four independent experiments. *p < 0.05 versus vehicle or 0 min (two-way ANOVA followed by Tukey–Kramer test).

Journal: The Journal of Neuroscience

Article Title: Pituitary Adenylate Cyclase-Activating Polypeptide Modulates Dendritic Spine Maturation and Morphogenesis via MicroRNA-132 Upregulation

doi: 10.1523/JNEUROSCI.2468-18.2019

Figure Lengend Snippet: PACAP significantly increased the number of function synapse in the primary cultured hippocampal neurons. A, Representative fluorescence images of the primary hippocampal neuronal dendrites at DIV21. The cells were fixed at DIV21 and immunostained using anti-synaptophysin (magenta) and anti-PSD95 (green) antibodies. Magnification of the area marked with a white dot box. Scale bar, 10 μm. B, Primary hippocampal neurons were treated for 2 d with 1, 10, or 100 nm PACAP. Scale bar, 10 μm. C, Quantification of the number of PSD-95-positive spines that were apposed to synaptophysin-positive presynaptic terminals. Values indicate mean ± SEM on 60 neurons from three independent experiments. **p < 0.01 versus vehicle (one-way ANOVA followed by Tukey–Kramer test). D, Immunoblot of cell surface biotinylated α1 sodium potassium ATPase, NR1, and NR2B from the primary cultured hippocampal neurons at DIV14. Surface biotinylation assay was performed after 2 d treatment with vehicle or 10 nm PACAP. E, Quantification of the cell surface expression levels of NR1 and NR2B were normalized to the levels of α1 sodium potassium ATPase. Values indicate mean ± SEM from five independent experiments. *p < 0.05 versus vehicle (Student's t test). F, Representative immunoblots showing expression of pERK1/2 and ERK2. Synaptic NMDAR activity in the primary cultured hippocampal neurons at DIV14 were treated with vehicle or 500 μm 4-AP and 50 μm bicuculline (bic) for 5 or 30 min after 2 d treatment with vehicle or 10 nm PACAP. 4-AP is a potassium channel blocker, and bicuculline is a GABAA receptor blocker. G, Quantification of the expression levels of pERK2 were normalized to the levels of ERK2. Values indicate mean ± SEM from four independent experiments. *p < 0.05 versus vehicle or 0 min (two-way ANOVA followed by Tukey–Kramer test).

Article Snippet: For predendritic and postdendritic spine labeling, the primary antibodies were used a rabbit polyclonal anti-PSD-95 antibody (1:500; catalog #3450S, RRID: AB_2292883 , Cell Signaling Technology) and a mouse monoclonal anti-synaptophysin antibody (1:2000; catalog #611880, RRID: AB_399360 , BD Biosciences) diluted in 0.2% Triton X-100 in PBS supplemented with 3% BSA.

Techniques: Cell Culture, Fluorescence, Western Blot, Surface Biotinylation Assay, Expressing, Activity Assay

Viral expression of miR-132 increased dendritic spine density in the primary hippocampal neurons from PACAP−/− mice. A, Representative fluorescence images of the primary hippocampal neuronal dendrites at DIV21. The cells were fixed at DIV21 and immunostained using anti-synaptophysin (magenta) and anti-PSD95 (green) antibodies. Scale bar, 10 μm. B, Quantification of the number of PSD-95-positive spines that were apposed to synaptophysin-positive presynaptic terminals. Values indicate mean ± SEM on 80 neurons from four independent experiments. *p < 0.05 versus vehicle (one-way ANOVA followed by Tukey–Kramer test). C, Expression levels of miR-132 in WT and PACAP−/− mice primary hippocampal neurons were analyzed by qRT-PCR. Primary hippocampal neurons were harvested at DIV20. Values indicate mean ± SEM from nine independent experiments. *p < 0.05 versus WT (one-way ANOVA followed by Tukey-kramer test). D, Representative fluorescence images of primary hippocampal neuron dendrites at DIV21. Primary hippocampal neurons were transduced with miR-132 Venus or control-Venus lentivirus at DIV14 and treated with 10 nm PACAP at DIV19. The cells were fixed at DIV21 and immunostained with anti-GFP (green) and anti-PSD95 (magenta) antibodies at DIV21. Scale bar, 10 μm. E–G, Quantification of the number of total spines (E), PSD-95-positive spines (F), and -negative spines (G). Values indicate mean ± SEM of 60 neurons from three independent experiments. **p < 0.01 versus Venus or WT (two-way ANOVA followed by Tukey–Kramer test).

Journal: The Journal of Neuroscience

Article Title: Pituitary Adenylate Cyclase-Activating Polypeptide Modulates Dendritic Spine Maturation and Morphogenesis via MicroRNA-132 Upregulation

doi: 10.1523/JNEUROSCI.2468-18.2019

Figure Lengend Snippet: Viral expression of miR-132 increased dendritic spine density in the primary hippocampal neurons from PACAP−/− mice. A, Representative fluorescence images of the primary hippocampal neuronal dendrites at DIV21. The cells were fixed at DIV21 and immunostained using anti-synaptophysin (magenta) and anti-PSD95 (green) antibodies. Scale bar, 10 μm. B, Quantification of the number of PSD-95-positive spines that were apposed to synaptophysin-positive presynaptic terminals. Values indicate mean ± SEM on 80 neurons from four independent experiments. *p < 0.05 versus vehicle (one-way ANOVA followed by Tukey–Kramer test). C, Expression levels of miR-132 in WT and PACAP−/− mice primary hippocampal neurons were analyzed by qRT-PCR. Primary hippocampal neurons were harvested at DIV20. Values indicate mean ± SEM from nine independent experiments. *p < 0.05 versus WT (one-way ANOVA followed by Tukey-kramer test). D, Representative fluorescence images of primary hippocampal neuron dendrites at DIV21. Primary hippocampal neurons were transduced with miR-132 Venus or control-Venus lentivirus at DIV14 and treated with 10 nm PACAP at DIV19. The cells were fixed at DIV21 and immunostained with anti-GFP (green) and anti-PSD95 (magenta) antibodies at DIV21. Scale bar, 10 μm. E–G, Quantification of the number of total spines (E), PSD-95-positive spines (F), and -negative spines (G). Values indicate mean ± SEM of 60 neurons from three independent experiments. **p < 0.01 versus Venus or WT (two-way ANOVA followed by Tukey–Kramer test).

Article Snippet: For predendritic and postdendritic spine labeling, the primary antibodies were used a rabbit polyclonal anti-PSD-95 antibody (1:500; catalog #3450S, RRID: AB_2292883 , Cell Signaling Technology) and a mouse monoclonal anti-synaptophysin antibody (1:2000; catalog #611880, RRID: AB_399360 , BD Biosciences) diluted in 0.2% Triton X-100 in PBS supplemented with 3% BSA.

Techniques: Expressing, Fluorescence, Quantitative RT-PCR, Transduction