pre cooled immunoelectron microscopy fixative Search Results


rabbit anti-synaptojanin1  (Synaptic Systems)
90
Synaptic Systems rabbit anti-synaptojanin1
Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, Endophilin1-, and <t>Synaptojanin1</t> puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.
Rabbit Anti Synaptojanin1, supplied by Synaptic Systems, 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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rabbit anti-dynamin3  (Synaptic Systems)
90
Synaptic Systems rabbit anti-dynamin3
Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against <t>Dynamin3.</t> Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, Endophilin1-, and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.
Rabbit Anti Dynamin3, supplied by Synaptic Systems, 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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polyclonal rabbit anti-amphiphysin  (Synaptic Systems)
90
Synaptic Systems polyclonal rabbit anti-amphiphysin
Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against <t>Dynamin3.</t> Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, Endophilin1-, and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.
Polyclonal Rabbit Anti Amphiphysin, supplied by Synaptic Systems, 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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rabbit anti-endophilin1  (Synaptic Systems)
90
Synaptic Systems rabbit anti-endophilin1
Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, <t>Endophilin1-,</t> and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.
Rabbit Anti Endophilin1, supplied by Synaptic Systems, 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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immunoelectron microscopic observation  (Federation of European Neuroscience Societies)
90
Federation of European Neuroscience Societies immunoelectron microscopic observation
Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, <t>Endophilin1-,</t> and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.
Immunoelectron Microscopic Observation, supplied by Federation of European Neuroscience Societies, 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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immunoelectron microscopy (iem)  (Johns Hopkins HealthCare)
90
Johns Hopkins HealthCare immunoelectron microscopy (iem)
Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, <t>Endophilin1-,</t> and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.
Immunoelectron Microscopy (Iem), supplied by Johns Hopkins HealthCare, 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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immunoelectron microscopy  (ACADEMIC PRESS INC)
90
ACADEMIC PRESS INC immunoelectron microscopy
Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, <t>Endophilin1-,</t> and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.
Immunoelectron Microscopy, supplied by ACADEMIC PRESS 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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5-nm-colloidal-immunogold-labeled anti-mouse goat igg  (EY Laboratories)
90
EY Laboratories 5-nm-colloidal-immunogold-labeled anti-mouse goat igg
Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, <t>Endophilin1-,</t> and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.
5 Nm Colloidal Immunogold Labeled Anti Mouse Goat Igg, supplied by EY Laboratories, 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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immunoelectron microscope fixative  (Servicebio Inc)
86
Servicebio Inc immunoelectron microscope fixative
Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, <t>Endophilin1-,</t> and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.
Immunoelectron Microscope Fixative, supplied by Servicebio Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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immunoelectron microscopy  (Hamamatsu)
90
Hamamatsu immunoelectron microscopy
Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, <t>Endophilin1-,</t> and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.
Immunoelectron Microscopy, supplied by Hamamatsu, 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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immunoelectronnance  (Vorum Research Corporation)
90
Vorum Research Corporation immunoelectronnance
Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, <t>Endophilin1-,</t> and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.
Immunoelectronnance, supplied by Vorum Research Corporation, 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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immunoelectron microscopy  (Federation of European Neuroscience Societies)
90
Federation of European Neuroscience Societies immunoelectron microscopy
Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, <t>Endophilin1-,</t> and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.
Immunoelectron Microscopy, supplied by Federation of European Neuroscience Societies, 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


Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, Endophilin1-, and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.

Journal: Frontiers in Cellular Neuroscience

Article Title: Evidence for a Clathrin-independent mode of endocytosis at a continuously active sensory synapse

doi: 10.3389/fncel.2014.00060

Figure Lengend Snippet: Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, Endophilin1-, and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.

Article Snippet: The following antibodies were used for ICC and pre-embedding immunoelectron microscopy (pre-EM): monoclonal mouse anti-Calbindin (ICC 1:2,000; #214011 Synaptic Systems, Göttingen, Germany), mouse anti-Clathrin light chain (Clathrin LC; ICC 1:1,000; #113011 Synaptic Systems), mouse anti-Dynamin (ICC 1:10,000; #ADI-VAM-SV041-E Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-uncoating ATPase (hsc70; ICC 1:100; #149011 Synaptic Systems), polyclonal rabbit anti-Amphiphysin (ICC/pre-EM 1:10,000; #120002 Synaptic Systems), rabbit anti-AP180 (ICC 1:10,000; #155003 Synaptic Systems), rabbit anti-Caveolin1 (ICC 1:1000; #161003 Synaptic Systems), rabbit anti-Dynamin3 (ICC/pre-EM 1:10,000; #115302 Synaptic Systems), rabbit anti-Endophilin1 (ICC/pre-EM 1:10,000; #159002 Synaptic Systems), rabbit anti-Synaptojanin1 (ICC 1:5,000; pre-EM 1:10,000; #145003 Synaptic Systems), rabbit anti-RIBEYE A-domain (ICC 1:50,000; #192103 Synaptic Systems; ), rabbit anti-Velis-3 (ICC 1:5,000, #51-5600 Invitrogen/Life technologies, Carlsbad, CA, USA; ), guinea pig anti-GFP (ICC 1:400; this antibody was generated and affinity-purified as described in .

Techniques: Labeling, Staining

Colocalization of Dynamin3 with Endophilin1 and Synaptojanin1 in 15 min, 3 h, and 15 h dark adapted OPL. (A,B) Outer plexiform layer (OPL) of C57BL/6JRj retinae after 3 h of dark adaptation labeled with antibodies against Dynamin3 (green) and Endophilin1 (A) or Synaptojanin1 ( B ; magenta). Arrows in the high power views point to individual, non-colocalizing puncta. (C) Quantification of the colocalization of Dynamin3 puncta with Endophilin1 and Synaptojanin1 in the OPL after 15 min, 3 h, and 15 h of dark adaptation. Values are means ± SD ( n = 3 retinae from 3 mice, 3 images per retina). Scale bar in B (for A,B ): 5 μm.

Journal: Frontiers in Cellular Neuroscience

Article Title: Evidence for a Clathrin-independent mode of endocytosis at a continuously active sensory synapse

doi: 10.3389/fncel.2014.00060

Figure Lengend Snippet: Colocalization of Dynamin3 with Endophilin1 and Synaptojanin1 in 15 min, 3 h, and 15 h dark adapted OPL. (A,B) Outer plexiform layer (OPL) of C57BL/6JRj retinae after 3 h of dark adaptation labeled with antibodies against Dynamin3 (green) and Endophilin1 (A) or Synaptojanin1 ( B ; magenta). Arrows in the high power views point to individual, non-colocalizing puncta. (C) Quantification of the colocalization of Dynamin3 puncta with Endophilin1 and Synaptojanin1 in the OPL after 15 min, 3 h, and 15 h of dark adaptation. Values are means ± SD ( n = 3 retinae from 3 mice, 3 images per retina). Scale bar in B (for A,B ): 5 μm.

Article Snippet: The following antibodies were used for ICC and pre-embedding immunoelectron microscopy (pre-EM): monoclonal mouse anti-Calbindin (ICC 1:2,000; #214011 Synaptic Systems, Göttingen, Germany), mouse anti-Clathrin light chain (Clathrin LC; ICC 1:1,000; #113011 Synaptic Systems), mouse anti-Dynamin (ICC 1:10,000; #ADI-VAM-SV041-E Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-uncoating ATPase (hsc70; ICC 1:100; #149011 Synaptic Systems), polyclonal rabbit anti-Amphiphysin (ICC/pre-EM 1:10,000; #120002 Synaptic Systems), rabbit anti-AP180 (ICC 1:10,000; #155003 Synaptic Systems), rabbit anti-Caveolin1 (ICC 1:1000; #161003 Synaptic Systems), rabbit anti-Dynamin3 (ICC/pre-EM 1:10,000; #115302 Synaptic Systems), rabbit anti-Endophilin1 (ICC/pre-EM 1:10,000; #159002 Synaptic Systems), rabbit anti-Synaptojanin1 (ICC 1:5,000; pre-EM 1:10,000; #145003 Synaptic Systems), rabbit anti-RIBEYE A-domain (ICC 1:50,000; #192103 Synaptic Systems; ), rabbit anti-Velis-3 (ICC 1:5,000, #51-5600 Invitrogen/Life technologies, Carlsbad, CA, USA; ), guinea pig anti-GFP (ICC 1:400; this antibody was generated and affinity-purified as described in .

Techniques: Labeling

Presynaptic localization of Dynamin3, Endophilin1, and Synaptojanin1 in 3 h dark adapted rod photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retina after 3 h of dark adaptation labeled with antibodies against Calbindin (green), Dynamin3 (red), and Piccolino (blue). Arrows point to adjacent Calbindin- and Dynamin3-puncta. (B–E) Pre-embedding immunoelectron micrographs of 3 h dark adapted rod photoreceptor terminals stained with antibodies against Dynamin3 (B,C) , Endophilin1 (D) , and Synaptojanin1 (E) . Immunoreactivity is encircled with dashed lines; arrowheads point to synaptic ribbons. HC, horizontal cell; BC, bipolar cell. Scale bar in A : 5 μm, in D (for B,D,E ): 200 nm, in C : 100 nm.

Journal: Frontiers in Cellular Neuroscience

Article Title: Evidence for a Clathrin-independent mode of endocytosis at a continuously active sensory synapse

doi: 10.3389/fncel.2014.00060

Figure Lengend Snippet: Presynaptic localization of Dynamin3, Endophilin1, and Synaptojanin1 in 3 h dark adapted rod photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retina after 3 h of dark adaptation labeled with antibodies against Calbindin (green), Dynamin3 (red), and Piccolino (blue). Arrows point to adjacent Calbindin- and Dynamin3-puncta. (B–E) Pre-embedding immunoelectron micrographs of 3 h dark adapted rod photoreceptor terminals stained with antibodies against Dynamin3 (B,C) , Endophilin1 (D) , and Synaptojanin1 (E) . Immunoreactivity is encircled with dashed lines; arrowheads point to synaptic ribbons. HC, horizontal cell; BC, bipolar cell. Scale bar in A : 5 μm, in D (for B,D,E ): 200 nm, in C : 100 nm.

Article Snippet: The following antibodies were used for ICC and pre-embedding immunoelectron microscopy (pre-EM): monoclonal mouse anti-Calbindin (ICC 1:2,000; #214011 Synaptic Systems, Göttingen, Germany), mouse anti-Clathrin light chain (Clathrin LC; ICC 1:1,000; #113011 Synaptic Systems), mouse anti-Dynamin (ICC 1:10,000; #ADI-VAM-SV041-E Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-uncoating ATPase (hsc70; ICC 1:100; #149011 Synaptic Systems), polyclonal rabbit anti-Amphiphysin (ICC/pre-EM 1:10,000; #120002 Synaptic Systems), rabbit anti-AP180 (ICC 1:10,000; #155003 Synaptic Systems), rabbit anti-Caveolin1 (ICC 1:1000; #161003 Synaptic Systems), rabbit anti-Dynamin3 (ICC/pre-EM 1:10,000; #115302 Synaptic Systems), rabbit anti-Endophilin1 (ICC/pre-EM 1:10,000; #159002 Synaptic Systems), rabbit anti-Synaptojanin1 (ICC 1:5,000; pre-EM 1:10,000; #145003 Synaptic Systems), rabbit anti-RIBEYE A-domain (ICC 1:50,000; #192103 Synaptic Systems; ), rabbit anti-Velis-3 (ICC 1:5,000, #51-5600 Invitrogen/Life technologies, Carlsbad, CA, USA; ), guinea pig anti-GFP (ICC 1:400; this antibody was generated and affinity-purified as described in .

Techniques: Labeling, Staining

Schematic summary of the hypothesized light/activity-dependent modes of endocytosis in photoreceptor terminals. (A–C) Photoreceptor ribbon synaptic site showing the postulated modes of endocytosis and the distribution of Dynamin3, Endophilin1, Synaptojanin1, and Clathrin LC (represented as colored triangles) in the light adapted state (A) , after short periods of darkness (1 to 15 min; B ), and after prolonged periods of darkness (3 to 15 h; C ). HC, horizontal cell; BC, bipolar cell.

Journal: Frontiers in Cellular Neuroscience

Article Title: Evidence for a Clathrin-independent mode of endocytosis at a continuously active sensory synapse

doi: 10.3389/fncel.2014.00060

Figure Lengend Snippet: Schematic summary of the hypothesized light/activity-dependent modes of endocytosis in photoreceptor terminals. (A–C) Photoreceptor ribbon synaptic site showing the postulated modes of endocytosis and the distribution of Dynamin3, Endophilin1, Synaptojanin1, and Clathrin LC (represented as colored triangles) in the light adapted state (A) , after short periods of darkness (1 to 15 min; B ), and after prolonged periods of darkness (3 to 15 h; C ). HC, horizontal cell; BC, bipolar cell.

Article Snippet: The following antibodies were used for ICC and pre-embedding immunoelectron microscopy (pre-EM): monoclonal mouse anti-Calbindin (ICC 1:2,000; #214011 Synaptic Systems, Göttingen, Germany), mouse anti-Clathrin light chain (Clathrin LC; ICC 1:1,000; #113011 Synaptic Systems), mouse anti-Dynamin (ICC 1:10,000; #ADI-VAM-SV041-E Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-uncoating ATPase (hsc70; ICC 1:100; #149011 Synaptic Systems), polyclonal rabbit anti-Amphiphysin (ICC/pre-EM 1:10,000; #120002 Synaptic Systems), rabbit anti-AP180 (ICC 1:10,000; #155003 Synaptic Systems), rabbit anti-Caveolin1 (ICC 1:1000; #161003 Synaptic Systems), rabbit anti-Dynamin3 (ICC/pre-EM 1:10,000; #115302 Synaptic Systems), rabbit anti-Endophilin1 (ICC/pre-EM 1:10,000; #159002 Synaptic Systems), rabbit anti-Synaptojanin1 (ICC 1:5,000; pre-EM 1:10,000; #145003 Synaptic Systems), rabbit anti-RIBEYE A-domain (ICC 1:50,000; #192103 Synaptic Systems; ), rabbit anti-Velis-3 (ICC 1:5,000, #51-5600 Invitrogen/Life technologies, Carlsbad, CA, USA; ), guinea pig anti-GFP (ICC 1:400; this antibody was generated and affinity-purified as described in .

Techniques: Activity Assay

Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, Endophilin1-, and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.

Journal: Frontiers in Cellular Neuroscience

Article Title: Evidence for a Clathrin-independent mode of endocytosis at a continuously active sensory synapse

doi: 10.3389/fncel.2014.00060

Figure Lengend Snippet: Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, Endophilin1-, and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.

Article Snippet: The following antibodies were used for ICC and pre-embedding immunoelectron microscopy (pre-EM): monoclonal mouse anti-Calbindin (ICC 1:2,000; #214011 Synaptic Systems, Göttingen, Germany), mouse anti-Clathrin light chain (Clathrin LC; ICC 1:1,000; #113011 Synaptic Systems), mouse anti-Dynamin (ICC 1:10,000; #ADI-VAM-SV041-E Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-uncoating ATPase (hsc70; ICC 1:100; #149011 Synaptic Systems), polyclonal rabbit anti-Amphiphysin (ICC/pre-EM 1:10,000; #120002 Synaptic Systems), rabbit anti-AP180 (ICC 1:10,000; #155003 Synaptic Systems), rabbit anti-Caveolin1 (ICC 1:1000; #161003 Synaptic Systems), rabbit anti-Dynamin3 (ICC/pre-EM 1:10,000; #115302 Synaptic Systems), rabbit anti-Endophilin1 (ICC/pre-EM 1:10,000; #159002 Synaptic Systems), rabbit anti-Synaptojanin1 (ICC 1:5,000; pre-EM 1:10,000; #145003 Synaptic Systems), rabbit anti-RIBEYE A-domain (ICC 1:50,000; #192103 Synaptic Systems; ), rabbit anti-Velis-3 (ICC 1:5,000, #51-5600 Invitrogen/Life technologies, Carlsbad, CA, USA; ), guinea pig anti-GFP (ICC 1:400; this antibody was generated and affinity-purified as described in .

Techniques: Labeling, Staining

Colocalization of Dynamin3 with Endophilin1 and Synaptojanin1 in 15 min, 3 h, and 15 h dark adapted OPL. (A,B) Outer plexiform layer (OPL) of C57BL/6JRj retinae after 3 h of dark adaptation labeled with antibodies against Dynamin3 (green) and Endophilin1 (A) or Synaptojanin1 ( B ; magenta). Arrows in the high power views point to individual, non-colocalizing puncta. (C) Quantification of the colocalization of Dynamin3 puncta with Endophilin1 and Synaptojanin1 in the OPL after 15 min, 3 h, and 15 h of dark adaptation. Values are means ± SD ( n = 3 retinae from 3 mice, 3 images per retina). Scale bar in B (for A,B ): 5 μm.

Journal: Frontiers in Cellular Neuroscience

Article Title: Evidence for a Clathrin-independent mode of endocytosis at a continuously active sensory synapse

doi: 10.3389/fncel.2014.00060

Figure Lengend Snippet: Colocalization of Dynamin3 with Endophilin1 and Synaptojanin1 in 15 min, 3 h, and 15 h dark adapted OPL. (A,B) Outer plexiform layer (OPL) of C57BL/6JRj retinae after 3 h of dark adaptation labeled with antibodies against Dynamin3 (green) and Endophilin1 (A) or Synaptojanin1 ( B ; magenta). Arrows in the high power views point to individual, non-colocalizing puncta. (C) Quantification of the colocalization of Dynamin3 puncta with Endophilin1 and Synaptojanin1 in the OPL after 15 min, 3 h, and 15 h of dark adaptation. Values are means ± SD ( n = 3 retinae from 3 mice, 3 images per retina). Scale bar in B (for A,B ): 5 μm.

Article Snippet: The following antibodies were used for ICC and pre-embedding immunoelectron microscopy (pre-EM): monoclonal mouse anti-Calbindin (ICC 1:2,000; #214011 Synaptic Systems, Göttingen, Germany), mouse anti-Clathrin light chain (Clathrin LC; ICC 1:1,000; #113011 Synaptic Systems), mouse anti-Dynamin (ICC 1:10,000; #ADI-VAM-SV041-E Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-uncoating ATPase (hsc70; ICC 1:100; #149011 Synaptic Systems), polyclonal rabbit anti-Amphiphysin (ICC/pre-EM 1:10,000; #120002 Synaptic Systems), rabbit anti-AP180 (ICC 1:10,000; #155003 Synaptic Systems), rabbit anti-Caveolin1 (ICC 1:1000; #161003 Synaptic Systems), rabbit anti-Dynamin3 (ICC/pre-EM 1:10,000; #115302 Synaptic Systems), rabbit anti-Endophilin1 (ICC/pre-EM 1:10,000; #159002 Synaptic Systems), rabbit anti-Synaptojanin1 (ICC 1:5,000; pre-EM 1:10,000; #145003 Synaptic Systems), rabbit anti-RIBEYE A-domain (ICC 1:50,000; #192103 Synaptic Systems; ), rabbit anti-Velis-3 (ICC 1:5,000, #51-5600 Invitrogen/Life technologies, Carlsbad, CA, USA; ), guinea pig anti-GFP (ICC 1:400; this antibody was generated and affinity-purified as described in .

Techniques: Labeling

Presynaptic localization of Dynamin3, Endophilin1, and Synaptojanin1 in 3 h dark adapted rod photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retina after 3 h of dark adaptation labeled with antibodies against Calbindin (green), Dynamin3 (red), and Piccolino (blue). Arrows point to adjacent Calbindin- and Dynamin3-puncta. (B–E) Pre-embedding immunoelectron micrographs of 3 h dark adapted rod photoreceptor terminals stained with antibodies against Dynamin3 (B,C) , Endophilin1 (D) , and Synaptojanin1 (E) . Immunoreactivity is encircled with dashed lines; arrowheads point to synaptic ribbons. HC, horizontal cell; BC, bipolar cell. Scale bar in A : 5 μm, in D (for B,D,E ): 200 nm, in C : 100 nm.

Journal: Frontiers in Cellular Neuroscience

Article Title: Evidence for a Clathrin-independent mode of endocytosis at a continuously active sensory synapse

doi: 10.3389/fncel.2014.00060

Figure Lengend Snippet: Presynaptic localization of Dynamin3, Endophilin1, and Synaptojanin1 in 3 h dark adapted rod photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retina after 3 h of dark adaptation labeled with antibodies against Calbindin (green), Dynamin3 (red), and Piccolino (blue). Arrows point to adjacent Calbindin- and Dynamin3-puncta. (B–E) Pre-embedding immunoelectron micrographs of 3 h dark adapted rod photoreceptor terminals stained with antibodies against Dynamin3 (B,C) , Endophilin1 (D) , and Synaptojanin1 (E) . Immunoreactivity is encircled with dashed lines; arrowheads point to synaptic ribbons. HC, horizontal cell; BC, bipolar cell. Scale bar in A : 5 μm, in D (for B,D,E ): 200 nm, in C : 100 nm.

Article Snippet: The following antibodies were used for ICC and pre-embedding immunoelectron microscopy (pre-EM): monoclonal mouse anti-Calbindin (ICC 1:2,000; #214011 Synaptic Systems, Göttingen, Germany), mouse anti-Clathrin light chain (Clathrin LC; ICC 1:1,000; #113011 Synaptic Systems), mouse anti-Dynamin (ICC 1:10,000; #ADI-VAM-SV041-E Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-uncoating ATPase (hsc70; ICC 1:100; #149011 Synaptic Systems), polyclonal rabbit anti-Amphiphysin (ICC/pre-EM 1:10,000; #120002 Synaptic Systems), rabbit anti-AP180 (ICC 1:10,000; #155003 Synaptic Systems), rabbit anti-Caveolin1 (ICC 1:1000; #161003 Synaptic Systems), rabbit anti-Dynamin3 (ICC/pre-EM 1:10,000; #115302 Synaptic Systems), rabbit anti-Endophilin1 (ICC/pre-EM 1:10,000; #159002 Synaptic Systems), rabbit anti-Synaptojanin1 (ICC 1:5,000; pre-EM 1:10,000; #145003 Synaptic Systems), rabbit anti-RIBEYE A-domain (ICC 1:50,000; #192103 Synaptic Systems; ), rabbit anti-Velis-3 (ICC 1:5,000, #51-5600 Invitrogen/Life technologies, Carlsbad, CA, USA; ), guinea pig anti-GFP (ICC 1:400; this antibody was generated and affinity-purified as described in .

Techniques: Labeling, Staining

Clusters of electron-dense endocytotic vesicles in 15 h dark adapted BsnΔEx4/5 photoreceptor terminals. (A) Outer plexiform layer (OPL) of a 15 h dark adapted BsnΔEx4/5 retina labeled with an antibody against Dynamin3. (B,C) Electron micrographs of rod photoreceptor terminals in a 15 h dark adapted BsnΔEx4/5 retina. Clusters of electron dense endocytotic vesicles (CEV) are encircled with dashed lines; arrowheads point to synaptic ribbons. HC, horizontal cell. Scale bar in A : 10 μm, in C (for B,C ): 200 nm.

Journal: Frontiers in Cellular Neuroscience

Article Title: Evidence for a Clathrin-independent mode of endocytosis at a continuously active sensory synapse

doi: 10.3389/fncel.2014.00060

Figure Lengend Snippet: Clusters of electron-dense endocytotic vesicles in 15 h dark adapted BsnΔEx4/5 photoreceptor terminals. (A) Outer plexiform layer (OPL) of a 15 h dark adapted BsnΔEx4/5 retina labeled with an antibody against Dynamin3. (B,C) Electron micrographs of rod photoreceptor terminals in a 15 h dark adapted BsnΔEx4/5 retina. Clusters of electron dense endocytotic vesicles (CEV) are encircled with dashed lines; arrowheads point to synaptic ribbons. HC, horizontal cell. Scale bar in A : 10 μm, in C (for B,C ): 200 nm.

Article Snippet: The following antibodies were used for ICC and pre-embedding immunoelectron microscopy (pre-EM): monoclonal mouse anti-Calbindin (ICC 1:2,000; #214011 Synaptic Systems, Göttingen, Germany), mouse anti-Clathrin light chain (Clathrin LC; ICC 1:1,000; #113011 Synaptic Systems), mouse anti-Dynamin (ICC 1:10,000; #ADI-VAM-SV041-E Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-uncoating ATPase (hsc70; ICC 1:100; #149011 Synaptic Systems), polyclonal rabbit anti-Amphiphysin (ICC/pre-EM 1:10,000; #120002 Synaptic Systems), rabbit anti-AP180 (ICC 1:10,000; #155003 Synaptic Systems), rabbit anti-Caveolin1 (ICC 1:1000; #161003 Synaptic Systems), rabbit anti-Dynamin3 (ICC/pre-EM 1:10,000; #115302 Synaptic Systems), rabbit anti-Endophilin1 (ICC/pre-EM 1:10,000; #159002 Synaptic Systems), rabbit anti-Synaptojanin1 (ICC 1:5,000; pre-EM 1:10,000; #145003 Synaptic Systems), rabbit anti-RIBEYE A-domain (ICC 1:50,000; #192103 Synaptic Systems; ), rabbit anti-Velis-3 (ICC 1:5,000, #51-5600 Invitrogen/Life technologies, Carlsbad, CA, USA; ), guinea pig anti-GFP (ICC 1:400; this antibody was generated and affinity-purified as described in .

Techniques: Labeling

Schematic summary of the hypothesized light/activity-dependent modes of endocytosis in photoreceptor terminals. (A–C) Photoreceptor ribbon synaptic site showing the postulated modes of endocytosis and the distribution of Dynamin3, Endophilin1, Synaptojanin1, and Clathrin LC (represented as colored triangles) in the light adapted state (A) , after short periods of darkness (1 to 15 min; B ), and after prolonged periods of darkness (3 to 15 h; C ). HC, horizontal cell; BC, bipolar cell.

Journal: Frontiers in Cellular Neuroscience

Article Title: Evidence for a Clathrin-independent mode of endocytosis at a continuously active sensory synapse

doi: 10.3389/fncel.2014.00060

Figure Lengend Snippet: Schematic summary of the hypothesized light/activity-dependent modes of endocytosis in photoreceptor terminals. (A–C) Photoreceptor ribbon synaptic site showing the postulated modes of endocytosis and the distribution of Dynamin3, Endophilin1, Synaptojanin1, and Clathrin LC (represented as colored triangles) in the light adapted state (A) , after short periods of darkness (1 to 15 min; B ), and after prolonged periods of darkness (3 to 15 h; C ). HC, horizontal cell; BC, bipolar cell.

Article Snippet: The following antibodies were used for ICC and pre-embedding immunoelectron microscopy (pre-EM): monoclonal mouse anti-Calbindin (ICC 1:2,000; #214011 Synaptic Systems, Göttingen, Germany), mouse anti-Clathrin light chain (Clathrin LC; ICC 1:1,000; #113011 Synaptic Systems), mouse anti-Dynamin (ICC 1:10,000; #ADI-VAM-SV041-E Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-uncoating ATPase (hsc70; ICC 1:100; #149011 Synaptic Systems), polyclonal rabbit anti-Amphiphysin (ICC/pre-EM 1:10,000; #120002 Synaptic Systems), rabbit anti-AP180 (ICC 1:10,000; #155003 Synaptic Systems), rabbit anti-Caveolin1 (ICC 1:1000; #161003 Synaptic Systems), rabbit anti-Dynamin3 (ICC/pre-EM 1:10,000; #115302 Synaptic Systems), rabbit anti-Endophilin1 (ICC/pre-EM 1:10,000; #159002 Synaptic Systems), rabbit anti-Synaptojanin1 (ICC 1:5,000; pre-EM 1:10,000; #145003 Synaptic Systems), rabbit anti-RIBEYE A-domain (ICC 1:50,000; #192103 Synaptic Systems; ), rabbit anti-Velis-3 (ICC 1:5,000, #51-5600 Invitrogen/Life technologies, Carlsbad, CA, USA; ), guinea pig anti-GFP (ICC 1:400; this antibody was generated and affinity-purified as described in .

Techniques: Activity Assay

Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, Endophilin1-, and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.

Journal: Frontiers in Cellular Neuroscience

Article Title: Evidence for a Clathrin-independent mode of endocytosis at a continuously active sensory synapse

doi: 10.3389/fncel.2014.00060

Figure Lengend Snippet: Redistribution of endocytotic proteins in 1 min, 15 min, 3 h, and 15 h dark adapted photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retinae after different periods of dark adaptation labeled with an antibody against Dynamin3. Single synaptic terminals are shown in high magnification. (B) Quantification of immunofluorescent Dynamin3-, Endophilin1-, and Synaptojanin1 puncta in the OPL after different durations of dark adaptation. (C) Images of rod photoreceptor terminals of C57BL/6JRj retina after 3 h of dark adaptation stained with antibodies against VGluT1 (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). (D) Images of cone photoreceptor terminals of Rac3-EGFP retina after 15 h of dark adaptation stained with antibodies against GFP (green) and Dynamin3, Endophilin1, or Synaptojanin1 (magenta). Single synaptic terminals are delineated with dashed lines. Values in B are means ± SD ( n = 4 retinae from 4 mice, 3 images per retina). Scale bar in A : 10 μm, in D (for C,D ): 2 μm.

Article Snippet: The following antibodies were used for ICC and pre-embedding immunoelectron microscopy (pre-EM): monoclonal mouse anti-Calbindin (ICC 1:2,000; #214011 Synaptic Systems, Göttingen, Germany), mouse anti-Clathrin light chain (Clathrin LC; ICC 1:1,000; #113011 Synaptic Systems), mouse anti-Dynamin (ICC 1:10,000; #ADI-VAM-SV041-E Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-uncoating ATPase (hsc70; ICC 1:100; #149011 Synaptic Systems), polyclonal rabbit anti-Amphiphysin (ICC/pre-EM 1:10,000; #120002 Synaptic Systems), rabbit anti-AP180 (ICC 1:10,000; #155003 Synaptic Systems), rabbit anti-Caveolin1 (ICC 1:1000; #161003 Synaptic Systems), rabbit anti-Dynamin3 (ICC/pre-EM 1:10,000; #115302 Synaptic Systems), rabbit anti-Endophilin1 (ICC/pre-EM 1:10,000; #159002 Synaptic Systems), rabbit anti-Synaptojanin1 (ICC 1:5,000; pre-EM 1:10,000; #145003 Synaptic Systems), rabbit anti-RIBEYE A-domain (ICC 1:50,000; #192103 Synaptic Systems; ), rabbit anti-Velis-3 (ICC 1:5,000, #51-5600 Invitrogen/Life technologies, Carlsbad, CA, USA; ), guinea pig anti-GFP (ICC 1:400; this antibody was generated and affinity-purified as described in .

Techniques: Labeling, Staining

Colocalization of Dynamin3 with Endophilin1 and Synaptojanin1 in 15 min, 3 h, and 15 h dark adapted OPL. (A,B) Outer plexiform layer (OPL) of C57BL/6JRj retinae after 3 h of dark adaptation labeled with antibodies against Dynamin3 (green) and Endophilin1 (A) or Synaptojanin1 ( B ; magenta). Arrows in the high power views point to individual, non-colocalizing puncta. (C) Quantification of the colocalization of Dynamin3 puncta with Endophilin1 and Synaptojanin1 in the OPL after 15 min, 3 h, and 15 h of dark adaptation. Values are means ± SD ( n = 3 retinae from 3 mice, 3 images per retina). Scale bar in B (for A,B ): 5 μm.

Journal: Frontiers in Cellular Neuroscience

Article Title: Evidence for a Clathrin-independent mode of endocytosis at a continuously active sensory synapse

doi: 10.3389/fncel.2014.00060

Figure Lengend Snippet: Colocalization of Dynamin3 with Endophilin1 and Synaptojanin1 in 15 min, 3 h, and 15 h dark adapted OPL. (A,B) Outer plexiform layer (OPL) of C57BL/6JRj retinae after 3 h of dark adaptation labeled with antibodies against Dynamin3 (green) and Endophilin1 (A) or Synaptojanin1 ( B ; magenta). Arrows in the high power views point to individual, non-colocalizing puncta. (C) Quantification of the colocalization of Dynamin3 puncta with Endophilin1 and Synaptojanin1 in the OPL after 15 min, 3 h, and 15 h of dark adaptation. Values are means ± SD ( n = 3 retinae from 3 mice, 3 images per retina). Scale bar in B (for A,B ): 5 μm.

Article Snippet: The following antibodies were used for ICC and pre-embedding immunoelectron microscopy (pre-EM): monoclonal mouse anti-Calbindin (ICC 1:2,000; #214011 Synaptic Systems, Göttingen, Germany), mouse anti-Clathrin light chain (Clathrin LC; ICC 1:1,000; #113011 Synaptic Systems), mouse anti-Dynamin (ICC 1:10,000; #ADI-VAM-SV041-E Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-uncoating ATPase (hsc70; ICC 1:100; #149011 Synaptic Systems), polyclonal rabbit anti-Amphiphysin (ICC/pre-EM 1:10,000; #120002 Synaptic Systems), rabbit anti-AP180 (ICC 1:10,000; #155003 Synaptic Systems), rabbit anti-Caveolin1 (ICC 1:1000; #161003 Synaptic Systems), rabbit anti-Dynamin3 (ICC/pre-EM 1:10,000; #115302 Synaptic Systems), rabbit anti-Endophilin1 (ICC/pre-EM 1:10,000; #159002 Synaptic Systems), rabbit anti-Synaptojanin1 (ICC 1:5,000; pre-EM 1:10,000; #145003 Synaptic Systems), rabbit anti-RIBEYE A-domain (ICC 1:50,000; #192103 Synaptic Systems; ), rabbit anti-Velis-3 (ICC 1:5,000, #51-5600 Invitrogen/Life technologies, Carlsbad, CA, USA; ), guinea pig anti-GFP (ICC 1:400; this antibody was generated and affinity-purified as described in .

Techniques: Labeling

Presynaptic localization of Dynamin3, Endophilin1, and Synaptojanin1 in 3 h dark adapted rod photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retina after 3 h of dark adaptation labeled with antibodies against Calbindin (green), Dynamin3 (red), and Piccolino (blue). Arrows point to adjacent Calbindin- and Dynamin3-puncta. (B–E) Pre-embedding immunoelectron micrographs of 3 h dark adapted rod photoreceptor terminals stained with antibodies against Dynamin3 (B,C) , Endophilin1 (D) , and Synaptojanin1 (E) . Immunoreactivity is encircled with dashed lines; arrowheads point to synaptic ribbons. HC, horizontal cell; BC, bipolar cell. Scale bar in A : 5 μm, in D (for B,D,E ): 200 nm, in C : 100 nm.

Journal: Frontiers in Cellular Neuroscience

Article Title: Evidence for a Clathrin-independent mode of endocytosis at a continuously active sensory synapse

doi: 10.3389/fncel.2014.00060

Figure Lengend Snippet: Presynaptic localization of Dynamin3, Endophilin1, and Synaptojanin1 in 3 h dark adapted rod photoreceptor terminals. (A) Outer plexiform layer (OPL) of C57BL/6JRj retina after 3 h of dark adaptation labeled with antibodies against Calbindin (green), Dynamin3 (red), and Piccolino (blue). Arrows point to adjacent Calbindin- and Dynamin3-puncta. (B–E) Pre-embedding immunoelectron micrographs of 3 h dark adapted rod photoreceptor terminals stained with antibodies against Dynamin3 (B,C) , Endophilin1 (D) , and Synaptojanin1 (E) . Immunoreactivity is encircled with dashed lines; arrowheads point to synaptic ribbons. HC, horizontal cell; BC, bipolar cell. Scale bar in A : 5 μm, in D (for B,D,E ): 200 nm, in C : 100 nm.

Article Snippet: The following antibodies were used for ICC and pre-embedding immunoelectron microscopy (pre-EM): monoclonal mouse anti-Calbindin (ICC 1:2,000; #214011 Synaptic Systems, Göttingen, Germany), mouse anti-Clathrin light chain (Clathrin LC; ICC 1:1,000; #113011 Synaptic Systems), mouse anti-Dynamin (ICC 1:10,000; #ADI-VAM-SV041-E Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-uncoating ATPase (hsc70; ICC 1:100; #149011 Synaptic Systems), polyclonal rabbit anti-Amphiphysin (ICC/pre-EM 1:10,000; #120002 Synaptic Systems), rabbit anti-AP180 (ICC 1:10,000; #155003 Synaptic Systems), rabbit anti-Caveolin1 (ICC 1:1000; #161003 Synaptic Systems), rabbit anti-Dynamin3 (ICC/pre-EM 1:10,000; #115302 Synaptic Systems), rabbit anti-Endophilin1 (ICC/pre-EM 1:10,000; #159002 Synaptic Systems), rabbit anti-Synaptojanin1 (ICC 1:5,000; pre-EM 1:10,000; #145003 Synaptic Systems), rabbit anti-RIBEYE A-domain (ICC 1:50,000; #192103 Synaptic Systems; ), rabbit anti-Velis-3 (ICC 1:5,000, #51-5600 Invitrogen/Life technologies, Carlsbad, CA, USA; ), guinea pig anti-GFP (ICC 1:400; this antibody was generated and affinity-purified as described in .

Techniques: Labeling, Staining

Schematic summary of the hypothesized light/activity-dependent modes of endocytosis in photoreceptor terminals. (A–C) Photoreceptor ribbon synaptic site showing the postulated modes of endocytosis and the distribution of Dynamin3, Endophilin1, Synaptojanin1, and Clathrin LC (represented as colored triangles) in the light adapted state (A) , after short periods of darkness (1 to 15 min; B ), and after prolonged periods of darkness (3 to 15 h; C ). HC, horizontal cell; BC, bipolar cell.

Journal: Frontiers in Cellular Neuroscience

Article Title: Evidence for a Clathrin-independent mode of endocytosis at a continuously active sensory synapse

doi: 10.3389/fncel.2014.00060

Figure Lengend Snippet: Schematic summary of the hypothesized light/activity-dependent modes of endocytosis in photoreceptor terminals. (A–C) Photoreceptor ribbon synaptic site showing the postulated modes of endocytosis and the distribution of Dynamin3, Endophilin1, Synaptojanin1, and Clathrin LC (represented as colored triangles) in the light adapted state (A) , after short periods of darkness (1 to 15 min; B ), and after prolonged periods of darkness (3 to 15 h; C ). HC, horizontal cell; BC, bipolar cell.

Article Snippet: The following antibodies were used for ICC and pre-embedding immunoelectron microscopy (pre-EM): monoclonal mouse anti-Calbindin (ICC 1:2,000; #214011 Synaptic Systems, Göttingen, Germany), mouse anti-Clathrin light chain (Clathrin LC; ICC 1:1,000; #113011 Synaptic Systems), mouse anti-Dynamin (ICC 1:10,000; #ADI-VAM-SV041-E Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-uncoating ATPase (hsc70; ICC 1:100; #149011 Synaptic Systems), polyclonal rabbit anti-Amphiphysin (ICC/pre-EM 1:10,000; #120002 Synaptic Systems), rabbit anti-AP180 (ICC 1:10,000; #155003 Synaptic Systems), rabbit anti-Caveolin1 (ICC 1:1000; #161003 Synaptic Systems), rabbit anti-Dynamin3 (ICC/pre-EM 1:10,000; #115302 Synaptic Systems), rabbit anti-Endophilin1 (ICC/pre-EM 1:10,000; #159002 Synaptic Systems), rabbit anti-Synaptojanin1 (ICC 1:5,000; pre-EM 1:10,000; #145003 Synaptic Systems), rabbit anti-RIBEYE A-domain (ICC 1:50,000; #192103 Synaptic Systems; ), rabbit anti-Velis-3 (ICC 1:5,000, #51-5600 Invitrogen/Life technologies, Carlsbad, CA, USA; ), guinea pig anti-GFP (ICC 1:400; this antibody was generated and affinity-purified as described in .

Techniques: Activity Assay