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hybridization buffer  (Thermo Fisher)
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Thermo Fisher hybridization buffer
Hybridization Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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md tritc dextran  (Thermo Fisher)
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Thermo Fisher md tritc dextran
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dextran alexa fluor 488  (Thermo Fisher)
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k gluconate 134  (Thermo Fisher)
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kd fluorescein dextran  (Thermo Fisher)
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Thermo Fisher kd fluorescein dextran
Involvement of the PKA-catalyzed phosphorylation of tomosyn in neurotransmitter release from SCG neurons. (A) Incidence of synaptic coupling between the nontransfected pre- and postsynaptic neurons (control) and between the presynaptic neurons that were transfected with myc-tomosyn (WT), myc-tomosyn–S724A (S724A), myc-tomosyn–S724D (S724D), tomosyn siRNA, or scramble RNA and the nontransfected postsynaptic neurons. The incidence for each dish is averaged ( n = 4–9). The bar graph shows means ± SEM (*, P < 0.05 vs. nontransfected presynaptic neurons). (B) EPSPs from one representative experiment recorded from the synapses between presynaptic neurons that were transfected with the respective tomosyn expression vector and the nontransfected postsynaptic neurons. Means of EPSP amplitudes ( n = 4–8; *, P < 0.05 vs. presynaptic neurons that were transfected with myc-tomosyn [WT]). (C) Expression levels of various tomosyn mutants and the effect of tomosyn knockdown by the RNA interference method in SCG neurons. SCG neurons were comicroinjected with <t>fluorescein</t> dextran as a morphological marker along with the same various tomosyn mutant (left) or siRNA (right) expression vectors that were used in A and were cultured for 40 h. The transfected cells were identified by microinjected fluorescein dextran (green), and the extent of expression of various tomosyn mutants was examined by immunostaining of tomosyn (red). The cell nuclei and plasma membrane borders of the neighboring nontransfected postsynaptic neurons are marked with dashed lines. N, nucleus. Arrows indicate the presynaptic terminals. Bars, 10 μm. (bottom) Quantification of relative fluorescence intensities of tomosyn at the presynaptic terminal. The results shown are representative of three independent experiments. Error bars represent SEM.
Kd Fluorescein Dextran, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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sodium dextran sulfate  (Thermo Fisher)
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Thermo Fisher sodium dextran sulfate
Involvement of the PKA-catalyzed phosphorylation of tomosyn in neurotransmitter release from SCG neurons. (A) Incidence of synaptic coupling between the nontransfected pre- and postsynaptic neurons (control) and between the presynaptic neurons that were transfected with myc-tomosyn (WT), myc-tomosyn–S724A (S724A), myc-tomosyn–S724D (S724D), tomosyn siRNA, or scramble RNA and the nontransfected postsynaptic neurons. The incidence for each dish is averaged ( n = 4–9). The bar graph shows means ± SEM (*, P < 0.05 vs. nontransfected presynaptic neurons). (B) EPSPs from one representative experiment recorded from the synapses between presynaptic neurons that were transfected with the respective tomosyn expression vector and the nontransfected postsynaptic neurons. Means of EPSP amplitudes ( n = 4–8; *, P < 0.05 vs. presynaptic neurons that were transfected with myc-tomosyn [WT]). (C) Expression levels of various tomosyn mutants and the effect of tomosyn knockdown by the RNA interference method in SCG neurons. SCG neurons were comicroinjected with <t>fluorescein</t> dextran as a morphological marker along with the same various tomosyn mutant (left) or siRNA (right) expression vectors that were used in A and were cultured for 40 h. The transfected cells were identified by microinjected fluorescein dextran (green), and the extent of expression of various tomosyn mutants was examined by immunostaining of tomosyn (red). The cell nuclei and plasma membrane borders of the neighboring nontransfected postsynaptic neurons are marked with dashed lines. N, nucleus. Arrows indicate the presynaptic terminals. Bars, 10 μm. (bottom) Quantification of relative fluorescence intensities of tomosyn at the presynaptic terminal. The results shown are representative of three independent experiments. Error bars represent SEM.
Sodium Dextran Sulfate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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medium dmem  (Thermo Fisher)
99
Thermo Fisher medium dmem
Involvement of the PKA-catalyzed phosphorylation of tomosyn in neurotransmitter release from SCG neurons. (A) Incidence of synaptic coupling between the nontransfected pre- and postsynaptic neurons (control) and between the presynaptic neurons that were transfected with myc-tomosyn (WT), myc-tomosyn–S724A (S724A), myc-tomosyn–S724D (S724D), tomosyn siRNA, or scramble RNA and the nontransfected postsynaptic neurons. The incidence for each dish is averaged ( n = 4–9). The bar graph shows means ± SEM (*, P < 0.05 vs. nontransfected presynaptic neurons). (B) EPSPs from one representative experiment recorded from the synapses between presynaptic neurons that were transfected with the respective tomosyn expression vector and the nontransfected postsynaptic neurons. Means of EPSP amplitudes ( n = 4–8; *, P < 0.05 vs. presynaptic neurons that were transfected with myc-tomosyn [WT]). (C) Expression levels of various tomosyn mutants and the effect of tomosyn knockdown by the RNA interference method in SCG neurons. SCG neurons were comicroinjected with <t>fluorescein</t> dextran as a morphological marker along with the same various tomosyn mutant (left) or siRNA (right) expression vectors that were used in A and were cultured for 40 h. The transfected cells were identified by microinjected fluorescein dextran (green), and the extent of expression of various tomosyn mutants was examined by immunostaining of tomosyn (red). The cell nuclei and plasma membrane borders of the neighboring nontransfected postsynaptic neurons are marked with dashed lines. N, nucleus. Arrows indicate the presynaptic terminals. Bars, 10 μm. (bottom) Quantification of relative fluorescence intensities of tomosyn at the presynaptic terminal. The results shown are representative of three independent experiments. Error bars represent SEM.
Medium Dmem, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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hybridisation buffer  (Thermo Fisher)
99
Thermo Fisher hybridisation buffer
Involvement of the PKA-catalyzed phosphorylation of tomosyn in neurotransmitter release from SCG neurons. (A) Incidence of synaptic coupling between the nontransfected pre- and postsynaptic neurons (control) and between the presynaptic neurons that were transfected with myc-tomosyn (WT), myc-tomosyn–S724A (S724A), myc-tomosyn–S724D (S724D), tomosyn siRNA, or scramble RNA and the nontransfected postsynaptic neurons. The incidence for each dish is averaged ( n = 4–9). The bar graph shows means ± SEM (*, P < 0.05 vs. nontransfected presynaptic neurons). (B) EPSPs from one representative experiment recorded from the synapses between presynaptic neurons that were transfected with the respective tomosyn expression vector and the nontransfected postsynaptic neurons. Means of EPSP amplitudes ( n = 4–8; *, P < 0.05 vs. presynaptic neurons that were transfected with myc-tomosyn [WT]). (C) Expression levels of various tomosyn mutants and the effect of tomosyn knockdown by the RNA interference method in SCG neurons. SCG neurons were comicroinjected with <t>fluorescein</t> dextran as a morphological marker along with the same various tomosyn mutant (left) or siRNA (right) expression vectors that were used in A and were cultured for 40 h. The transfected cells were identified by microinjected fluorescein dextran (green), and the extent of expression of various tomosyn mutants was examined by immunostaining of tomosyn (red). The cell nuclei and plasma membrane borders of the neighboring nontransfected postsynaptic neurons are marked with dashed lines. N, nucleus. Arrows indicate the presynaptic terminals. Bars, 10 μm. (bottom) Quantification of relative fluorescence intensities of tomosyn at the presynaptic terminal. The results shown are representative of three independent experiments. Error bars represent SEM.
Hybridisation Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/dextran+invitrogen/Tween+20/pmc08758788-398-40-35
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dye  (Biotium)
93
Biotium dye
Involvement of the PKA-catalyzed phosphorylation of tomosyn in neurotransmitter release from SCG neurons. (A) Incidence of synaptic coupling between the nontransfected pre- and postsynaptic neurons (control) and between the presynaptic neurons that were transfected with myc-tomosyn (WT), myc-tomosyn–S724A (S724A), myc-tomosyn–S724D (S724D), tomosyn siRNA, or scramble RNA and the nontransfected postsynaptic neurons. The incidence for each dish is averaged ( n = 4–9). The bar graph shows means ± SEM (*, P < 0.05 vs. nontransfected presynaptic neurons). (B) EPSPs from one representative experiment recorded from the synapses between presynaptic neurons that were transfected with the respective tomosyn expression vector and the nontransfected postsynaptic neurons. Means of EPSP amplitudes ( n = 4–8; *, P < 0.05 vs. presynaptic neurons that were transfected with myc-tomosyn [WT]). (C) Expression levels of various tomosyn mutants and the effect of tomosyn knockdown by the RNA interference method in SCG neurons. SCG neurons were comicroinjected with <t>fluorescein</t> dextran as a morphological marker along with the same various tomosyn mutant (left) or siRNA (right) expression vectors that were used in A and were cultured for 40 h. The transfected cells were identified by microinjected fluorescein dextran (green), and the extent of expression of various tomosyn mutants was examined by immunostaining of tomosyn (red). The cell nuclei and plasma membrane borders of the neighboring nontransfected postsynaptic neurons are marked with dashed lines. N, nucleus. Arrows indicate the presynaptic terminals. Bars, 10 μm. (bottom) Quantification of relative fluorescence intensities of tomosyn at the presynaptic terminal. The results shown are representative of three independent experiments. Error bars represent SEM.
Dye, supplied by Biotium, 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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serum albumin dbsa  (Thermo Fisher)
99
Thermo Fisher serum albumin dbsa
Involvement of the PKA-catalyzed phosphorylation of tomosyn in neurotransmitter release from SCG neurons. (A) Incidence of synaptic coupling between the nontransfected pre- and postsynaptic neurons (control) and between the presynaptic neurons that were transfected with myc-tomosyn (WT), myc-tomosyn–S724A (S724A), myc-tomosyn–S724D (S724D), tomosyn siRNA, or scramble RNA and the nontransfected postsynaptic neurons. The incidence for each dish is averaged ( n = 4–9). The bar graph shows means ± SEM (*, P < 0.05 vs. nontransfected presynaptic neurons). (B) EPSPs from one representative experiment recorded from the synapses between presynaptic neurons that were transfected with the respective tomosyn expression vector and the nontransfected postsynaptic neurons. Means of EPSP amplitudes ( n = 4–8; *, P < 0.05 vs. presynaptic neurons that were transfected with myc-tomosyn [WT]). (C) Expression levels of various tomosyn mutants and the effect of tomosyn knockdown by the RNA interference method in SCG neurons. SCG neurons were comicroinjected with <t>fluorescein</t> dextran as a morphological marker along with the same various tomosyn mutant (left) or siRNA (right) expression vectors that were used in A and were cultured for 40 h. The transfected cells were identified by microinjected fluorescein dextran (green), and the extent of expression of various tomosyn mutants was examined by immunostaining of tomosyn (red). The cell nuclei and plasma membrane borders of the neighboring nontransfected postsynaptic neurons are marked with dashed lines. N, nucleus. Arrows indicate the presynaptic terminals. Bars, 10 μm. (bottom) Quantification of relative fluorescence intensities of tomosyn at the presynaptic terminal. The results shown are representative of three independent experiments. Error bars represent SEM.
Serum Albumin Dbsa, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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fetal bovine serum  (BioIVT Inc)
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BioIVT Inc fetal bovine serum
Involvement of the PKA-catalyzed phosphorylation of tomosyn in neurotransmitter release from SCG neurons. (A) Incidence of synaptic coupling between the nontransfected pre- and postsynaptic neurons (control) and between the presynaptic neurons that were transfected with myc-tomosyn (WT), myc-tomosyn–S724A (S724A), myc-tomosyn–S724D (S724D), tomosyn siRNA, or scramble RNA and the nontransfected postsynaptic neurons. The incidence for each dish is averaged ( n = 4–9). The bar graph shows means ± SEM (*, P < 0.05 vs. nontransfected presynaptic neurons). (B) EPSPs from one representative experiment recorded from the synapses between presynaptic neurons that were transfected with the respective tomosyn expression vector and the nontransfected postsynaptic neurons. Means of EPSP amplitudes ( n = 4–8; *, P < 0.05 vs. presynaptic neurons that were transfected with myc-tomosyn [WT]). (C) Expression levels of various tomosyn mutants and the effect of tomosyn knockdown by the RNA interference method in SCG neurons. SCG neurons were comicroinjected with <t>fluorescein</t> dextran as a morphological marker along with the same various tomosyn mutant (left) or siRNA (right) expression vectors that were used in A and were cultured for 40 h. The transfected cells were identified by microinjected fluorescein dextran (green), and the extent of expression of various tomosyn mutants was examined by immunostaining of tomosyn (red). The cell nuclei and plasma membrane borders of the neighboring nontransfected postsynaptic neurons are marked with dashed lines. N, nucleus. Arrows indicate the presynaptic terminals. Bars, 10 μm. (bottom) Quantification of relative fluorescence intensities of tomosyn at the presynaptic terminal. The results shown are representative of three independent experiments. Error bars represent SEM.
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70 kda dextran  (Thermo Fisher)
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Thermo Fisher 70 kda dextran
Involvement of the PKA-catalyzed phosphorylation of tomosyn in neurotransmitter release from SCG neurons. (A) Incidence of synaptic coupling between the nontransfected pre- and postsynaptic neurons (control) and between the presynaptic neurons that were transfected with myc-tomosyn (WT), myc-tomosyn–S724A (S724A), myc-tomosyn–S724D (S724D), tomosyn siRNA, or scramble RNA and the nontransfected postsynaptic neurons. The incidence for each dish is averaged ( n = 4–9). The bar graph shows means ± SEM (*, P < 0.05 vs. nontransfected presynaptic neurons). (B) EPSPs from one representative experiment recorded from the synapses between presynaptic neurons that were transfected with the respective tomosyn expression vector and the nontransfected postsynaptic neurons. Means of EPSP amplitudes ( n = 4–8; *, P < 0.05 vs. presynaptic neurons that were transfected with myc-tomosyn [WT]). (C) Expression levels of various tomosyn mutants and the effect of tomosyn knockdown by the RNA interference method in SCG neurons. SCG neurons were comicroinjected with <t>fluorescein</t> dextran as a morphological marker along with the same various tomosyn mutant (left) or siRNA (right) expression vectors that were used in A and were cultured for 40 h. The transfected cells were identified by microinjected fluorescein dextran (green), and the extent of expression of various tomosyn mutants was examined by immunostaining of tomosyn (red). The cell nuclei and plasma membrane borders of the neighboring nontransfected postsynaptic neurons are marked with dashed lines. N, nucleus. Arrows indicate the presynaptic terminals. Bars, 10 μm. (bottom) Quantification of relative fluorescence intensities of tomosyn at the presynaptic terminal. The results shown are representative of three independent experiments. Error bars represent SEM.
70 Kda Dextran, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Involvement of the PKA-catalyzed phosphorylation of tomosyn in neurotransmitter release from SCG neurons. (A) Incidence of synaptic coupling between the nontransfected pre- and postsynaptic neurons (control) and between the presynaptic neurons that were transfected with myc-tomosyn (WT), myc-tomosyn–S724A (S724A), myc-tomosyn–S724D (S724D), tomosyn siRNA, or scramble RNA and the nontransfected postsynaptic neurons. The incidence for each dish is averaged ( n = 4–9). The bar graph shows means ± SEM (*, P < 0.05 vs. nontransfected presynaptic neurons). (B) EPSPs from one representative experiment recorded from the synapses between presynaptic neurons that were transfected with the respective tomosyn expression vector and the nontransfected postsynaptic neurons. Means of EPSP amplitudes ( n = 4–8; *, P < 0.05 vs. presynaptic neurons that were transfected with myc-tomosyn [WT]). (C) Expression levels of various tomosyn mutants and the effect of tomosyn knockdown by the RNA interference method in SCG neurons. SCG neurons were comicroinjected with fluorescein dextran as a morphological marker along with the same various tomosyn mutant (left) or siRNA (right) expression vectors that were used in A and were cultured for 40 h. The transfected cells were identified by microinjected fluorescein dextran (green), and the extent of expression of various tomosyn mutants was examined by immunostaining of tomosyn (red). The cell nuclei and plasma membrane borders of the neighboring nontransfected postsynaptic neurons are marked with dashed lines. N, nucleus. Arrows indicate the presynaptic terminals. Bars, 10 μm. (bottom) Quantification of relative fluorescence intensities of tomosyn at the presynaptic terminal. The results shown are representative of three independent experiments. Error bars represent SEM.

Journal: The Journal of Cell Biology

Article Title: PKA-catalyzed phosphorylation of tomosyn and its implication in Ca 2+ -dependent exocytosis of neurotransmitter

doi: 10.1083/jcb.200504055

Figure Lengend Snippet: Involvement of the PKA-catalyzed phosphorylation of tomosyn in neurotransmitter release from SCG neurons. (A) Incidence of synaptic coupling between the nontransfected pre- and postsynaptic neurons (control) and between the presynaptic neurons that were transfected with myc-tomosyn (WT), myc-tomosyn–S724A (S724A), myc-tomosyn–S724D (S724D), tomosyn siRNA, or scramble RNA and the nontransfected postsynaptic neurons. The incidence for each dish is averaged ( n = 4–9). The bar graph shows means ± SEM (*, P < 0.05 vs. nontransfected presynaptic neurons). (B) EPSPs from one representative experiment recorded from the synapses between presynaptic neurons that were transfected with the respective tomosyn expression vector and the nontransfected postsynaptic neurons. Means of EPSP amplitudes ( n = 4–8; *, P < 0.05 vs. presynaptic neurons that were transfected with myc-tomosyn [WT]). (C) Expression levels of various tomosyn mutants and the effect of tomosyn knockdown by the RNA interference method in SCG neurons. SCG neurons were comicroinjected with fluorescein dextran as a morphological marker along with the same various tomosyn mutant (left) or siRNA (right) expression vectors that were used in A and were cultured for 40 h. The transfected cells were identified by microinjected fluorescein dextran (green), and the extent of expression of various tomosyn mutants was examined by immunostaining of tomosyn (red). The cell nuclei and plasma membrane borders of the neighboring nontransfected postsynaptic neurons are marked with dashed lines. N, nucleus. Arrows indicate the presynaptic terminals. Bars, 10 μm. (bottom) Quantification of relative fluorescence intensities of tomosyn at the presynaptic terminal. The results shown are representative of three independent experiments. Error bars represent SEM.

Article Snippet: Various tomosyn or PKA expression vectors and the tomosyn or PKA siRNA expression vector were microinjected into the nuclei of SCG neurons through a microglass pipette along with 10 kD fluorescein dextran (Invitrogen) as described previously ( ).

Techniques: Phospho-proteomics, Control, Transfection, Expressing, Plasmid Preparation, Knockdown, Marker, Mutagenesis, Cell Culture, Immunostaining, Clinical Proteomics, Membrane, Fluorescence