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  • 88
    Synaptic Systems rim2
    Reduced exocytosis in <t>RIM2−deficient</t> IHCs. ( A ) Representative capacitance increments (ΔC m ) of a RIM2α SKO IHC (gray) and a RIM2α con IHC (black) in response to 100-ms long-step depolarizations to −14 mV. ( B ) Average
    Rim2, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 88/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Syntaxin rim2
    Reduced exocytosis in <t>RIM2−deficient</t> IHCs. ( A ) Representative capacitance increments (ΔC m ) of a RIM2α SKO IHC (gray) and a RIM2α con IHC (black) in response to 100-ms long-step depolarizations to −14 mV. ( B ) Average
    Rim2, supplied by Syntaxin, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    GeneTex rabbit anti rim2
    The knockdown of spinal RIM1α expression relieves SNL-induced allodynia. A , B , Representative Western blot and statistical analysis (normalized to GAPDH) demonstrating that the intrathecal administration of RIM1α mRNA-targeting siRNA (RIM1α RNAi; 1, 3, and 5 μg; 10 μl; once daily for 4 d), but not missense siRNA (MS; 5 μg, 10 μl) or polyethylenimine (a transfection reagent, PEI; 10 μl), dose-dependently decreased spinal RIM1α expression of naive rat. Yet, all these treatments failed to affect the abundance of <t>RIM2</t> in the dorsal horn sample of naive rat. it, Implantation of an intrathecal catheter; IB, immunoblotting. RIM1α, one-way ANOVA, post hoc Tukey's test, F (6,35) = 22.96, p
    Rabbit Anti Rim2, supplied by GeneTex, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Synaptic Systems anti rim2 pabs
    The knockdown of spinal RIM1α expression relieves SNL-induced allodynia. A , B , Representative Western blot and statistical analysis (normalized to GAPDH) demonstrating that the intrathecal administration of RIM1α mRNA-targeting siRNA (RIM1α RNAi; 1, 3, and 5 μg; 10 μl; once daily for 4 d), but not missense siRNA (MS; 5 μg, 10 μl) or polyethylenimine (a transfection reagent, PEI; 10 μl), dose-dependently decreased spinal RIM1α expression of naive rat. Yet, all these treatments failed to affect the abundance of <t>RIM2</t> in the dorsal horn sample of naive rat. it, Implantation of an intrathecal catheter; IB, immunoblotting. RIM1α, one-way ANOVA, post hoc Tukey's test, F (6,35) = 22.96, p
    Anti Rim2 Pabs, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 85/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Synaptic Systems anti rim2
    Conditional knock-out of <t>RIM2</t> in rod photoreceptors is under way as early as 3 weeks of age. A , Forms of RIM1 and RIM2 found in the brain and retina of wild-type and transgenic RIM1ff/2ff mice. Expression of RIM 1α ≫ 1β in the retina, which is similar to the proportions in whole-brain lysates (probed with Ms monoclonal anti-RIM1). The floxed RIM2ff ) and therefore all of the RIM2 in the transgenic animal is RIM2-cfp, which is revealed as an upward shift in its migration compared with brain lysates from wild-type animals (the cfp tag is nonfluorescent in the retina). Relative RIM2 expression levels in the brain are RIM 2α > 2β ∼ 2α-splice variants (α-sp.) and, in the retina, 2α and 2α-sp are equally expressed, but 2β is not detected. The regional distribution of RIM1/2 was examined by immunostaining cryosections of retina ∼10 μm thick ( B – F ). B , Expression of RIM1 is largely confined to the IPL and this is demonstrated with a rabbit polyclonal anti-RIM1 from Synaptic Systems (left) and a mouse monoclonal against RIM1 from BD Transduction Laboratories (right). Scale bar, 25 μm. C , D ), demarcating the OPL. The section in D is from a LMOPcre + transgenic animal, which shows an intensely stained patch of Cre in the ONL that is designated as a Cre region (samples in C and D are from RIM1ff/2ff littermates, 3 weeks of age). Bottom overview cropped to the OPL is labeled “all probes” because the signals are derived from anti-RIM2 staining (rabbit polyclonal, Alexa Fluor488, shown in green), the cone marker PNA (linked to Alexa Fluor 568, shown in red), and anti-CtBP2 (Ms monoclonal, Alexa Fluor 648, shown in blue). E , F , Regions selected for presentation at higher resolution. The control condition presented in C and E and the “Cre-poor region” in F show that the RIM2 signal overlaps extensively with CtBP2/Ribeye (aqua). The “Cre-rich region” in F reveals a drop in RIM2 staining. G , H ) and Synaptotagmin1 (Syt1, ∼60 kDa) were used as presynaptic markers, whereas CtBP2 (∼45 kDa) in the context of Western analysis represents the transcription factor that is expressed throughout the retina. I , Graphical summary of the densitometry measurements from Western analysis. Only RIM2 was reduced (see text for values) and no apparent change in the intensity of the synaptic markers was witnessed at 3 weeks. Data are presented as mean ± SE.
    Anti Rim2, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Synaptic Systems rabbit anti rim2 znfinger
    Conditional knock-out of <t>RIM2</t> in rod photoreceptors is under way as early as 3 weeks of age. A , Forms of RIM1 and RIM2 found in the brain and retina of wild-type and transgenic RIM1ff/2ff mice. Expression of RIM 1α ≫ 1β in the retina, which is similar to the proportions in whole-brain lysates (probed with Ms monoclonal anti-RIM1). The floxed RIM2ff ) and therefore all of the RIM2 in the transgenic animal is RIM2-cfp, which is revealed as an upward shift in its migration compared with brain lysates from wild-type animals (the cfp tag is nonfluorescent in the retina). Relative RIM2 expression levels in the brain are RIM 2α > 2β ∼ 2α-splice variants (α-sp.) and, in the retina, 2α and 2α-sp are equally expressed, but 2β is not detected. The regional distribution of RIM1/2 was examined by immunostaining cryosections of retina ∼10 μm thick ( B – F ). B , Expression of RIM1 is largely confined to the IPL and this is demonstrated with a rabbit polyclonal anti-RIM1 from Synaptic Systems (left) and a mouse monoclonal against RIM1 from BD Transduction Laboratories (right). Scale bar, 25 μm. C , D ), demarcating the OPL. The section in D is from a LMOPcre + transgenic animal, which shows an intensely stained patch of Cre in the ONL that is designated as a Cre region (samples in C and D are from RIM1ff/2ff littermates, 3 weeks of age). Bottom overview cropped to the OPL is labeled “all probes” because the signals are derived from anti-RIM2 staining (rabbit polyclonal, Alexa Fluor488, shown in green), the cone marker PNA (linked to Alexa Fluor 568, shown in red), and anti-CtBP2 (Ms monoclonal, Alexa Fluor 648, shown in blue). E , F , Regions selected for presentation at higher resolution. The control condition presented in C and E and the “Cre-poor region” in F show that the RIM2 signal overlaps extensively with CtBP2/Ribeye (aqua). The “Cre-rich region” in F reveals a drop in RIM2 staining. G , H ) and Synaptotagmin1 (Syt1, ∼60 kDa) were used as presynaptic markers, whereas CtBP2 (∼45 kDa) in the context of Western analysis represents the transcription factor that is expressed throughout the retina. I , Graphical summary of the densitometry measurements from Western analysis. Only RIM2 was reduced (see text for values) and no apparent change in the intensity of the synaptic markers was witnessed at 3 weeks. Data are presented as mean ± SE.
    Rabbit Anti Rim2 Znfinger, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Santa Cruz Biotechnology goat anti rim2
    Conditional knock-out of <t>RIM2</t> in rod photoreceptors is under way as early as 3 weeks of age. A , Forms of RIM1 and RIM2 found in the brain and retina of wild-type and transgenic RIM1ff/2ff mice. Expression of RIM 1α ≫ 1β in the retina, which is similar to the proportions in whole-brain lysates (probed with Ms monoclonal anti-RIM1). The floxed RIM2ff ) and therefore all of the RIM2 in the transgenic animal is RIM2-cfp, which is revealed as an upward shift in its migration compared with brain lysates from wild-type animals (the cfp tag is nonfluorescent in the retina). Relative RIM2 expression levels in the brain are RIM 2α > 2β ∼ 2α-splice variants (α-sp.) and, in the retina, 2α and 2α-sp are equally expressed, but 2β is not detected. The regional distribution of RIM1/2 was examined by immunostaining cryosections of retina ∼10 μm thick ( B – F ). B , Expression of RIM1 is largely confined to the IPL and this is demonstrated with a rabbit polyclonal anti-RIM1 from Synaptic Systems (left) and a mouse monoclonal against RIM1 from BD Transduction Laboratories (right). Scale bar, 25 μm. C , D ), demarcating the OPL. The section in D is from a LMOPcre + transgenic animal, which shows an intensely stained patch of Cre in the ONL that is designated as a Cre region (samples in C and D are from RIM1ff/2ff littermates, 3 weeks of age). Bottom overview cropped to the OPL is labeled “all probes” because the signals are derived from anti-RIM2 staining (rabbit polyclonal, Alexa Fluor488, shown in green), the cone marker PNA (linked to Alexa Fluor 568, shown in red), and anti-CtBP2 (Ms monoclonal, Alexa Fluor 648, shown in blue). E , F , Regions selected for presentation at higher resolution. The control condition presented in C and E and the “Cre-poor region” in F show that the RIM2 signal overlaps extensively with CtBP2/Ribeye (aqua). The “Cre-rich region” in F reveals a drop in RIM2 staining. G , H ) and Synaptotagmin1 (Syt1, ∼60 kDa) were used as presynaptic markers, whereas CtBP2 (∼45 kDa) in the context of Western analysis represents the transcription factor that is expressed throughout the retina. I , Graphical summary of the densitometry measurements from Western analysis. Only RIM2 was reduced (see text for values) and no apparent change in the intensity of the synaptic markers was witnessed at 3 weeks. Data are presented as mean ± SE.
    Goat Anti Rim2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Synaptic Systems rabbit anti rim2
    Conditional knock-out of <t>RIM2</t> in rod photoreceptors is under way as early as 3 weeks of age. A , Forms of RIM1 and RIM2 found in the brain and retina of wild-type and transgenic RIM1ff/2ff mice. Expression of RIM 1α ≫ 1β in the retina, which is similar to the proportions in whole-brain lysates (probed with Ms monoclonal anti-RIM1). The floxed RIM2ff ) and therefore all of the RIM2 in the transgenic animal is RIM2-cfp, which is revealed as an upward shift in its migration compared with brain lysates from wild-type animals (the cfp tag is nonfluorescent in the retina). Relative RIM2 expression levels in the brain are RIM 2α > 2β ∼ 2α-splice variants (α-sp.) and, in the retina, 2α and 2α-sp are equally expressed, but 2β is not detected. The regional distribution of RIM1/2 was examined by immunostaining cryosections of retina ∼10 μm thick ( B – F ). B , Expression of RIM1 is largely confined to the IPL and this is demonstrated with a rabbit polyclonal anti-RIM1 from Synaptic Systems (left) and a mouse monoclonal against RIM1 from BD Transduction Laboratories (right). Scale bar, 25 μm. C , D ), demarcating the OPL. The section in D is from a LMOPcre + transgenic animal, which shows an intensely stained patch of Cre in the ONL that is designated as a Cre region (samples in C and D are from RIM1ff/2ff littermates, 3 weeks of age). Bottom overview cropped to the OPL is labeled “all probes” because the signals are derived from anti-RIM2 staining (rabbit polyclonal, Alexa Fluor488, shown in green), the cone marker PNA (linked to Alexa Fluor 568, shown in red), and anti-CtBP2 (Ms monoclonal, Alexa Fluor 648, shown in blue). E , F , Regions selected for presentation at higher resolution. The control condition presented in C and E and the “Cre-poor region” in F show that the RIM2 signal overlaps extensively with CtBP2/Ribeye (aqua). The “Cre-rich region” in F reveals a drop in RIM2 staining. G , H ) and Synaptotagmin1 (Syt1, ∼60 kDa) were used as presynaptic markers, whereas CtBP2 (∼45 kDa) in the context of Western analysis represents the transcription factor that is expressed throughout the retina. I , Graphical summary of the densitometry measurements from Western analysis. Only RIM2 was reduced (see text for values) and no apparent change in the intensity of the synaptic markers was witnessed at 3 weeks. Data are presented as mean ± SE.
    Rabbit Anti Rim2, supplied by Synaptic Systems, used in various techniques. Bioz Stars score: 86/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Reduced exocytosis in RIM2−deficient IHCs. ( A ) Representative capacitance increments (ΔC m ) of a RIM2α SKO IHC (gray) and a RIM2α con IHC (black) in response to 100-ms long-step depolarizations to −14 mV. ( B ) Average

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

    Article Title: Rab3-interacting molecules 2α and 2β promote the abundance of voltage-gated CaV1.3 Ca2+ channels at hair cell active zones

    doi: 10.1073/pnas.1417207112

    Figure Lengend Snippet: Reduced exocytosis in RIM2−deficient IHCs. ( A ) Representative capacitance increments (ΔC m ) of a RIM2α SKO IHC (gray) and a RIM2α con IHC (black) in response to 100-ms long-step depolarizations to −14 mV. ( B ) Average

    Article Snippet: Primary antibodies were mouse anti-CtBP2 (1:200, BD Biosciences), rabbit anti-GluA2/3 (1:200, Chemicon), rabbit anti-CaV 1.3 (1: 50, Alomone Labs), mouse anti-GluA2 (1:75 Chemicon), RIM2 (#140103 recognizing the PSD-95/discs large/ZO-1 (PDZ) domain, 1:200; Synaptic Systems), RIM1 (1:200; Synaptic Systems), RIM3 (1:100) , and Vglut3 (1:500; Synaptic Systems).

    Techniques: Immunohistochemistry, Mass Spectrometry

    RIM2α, RIM2β, and RIM3γ are present at IHC AZs of hearing mice. ( A ) Projections of confocal sections of a wild-type organ of Corti immunolabeled with antibodies to RIM2 (green), CtBP2/RIBEYE (red, antibody labeling the presynaptic

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

    Article Title: Rab3-interacting molecules 2α and 2β promote the abundance of voltage-gated CaV1.3 Ca2+ channels at hair cell active zones

    doi: 10.1073/pnas.1417207112

    Figure Lengend Snippet: RIM2α, RIM2β, and RIM3γ are present at IHC AZs of hearing mice. ( A ) Projections of confocal sections of a wild-type organ of Corti immunolabeled with antibodies to RIM2 (green), CtBP2/RIBEYE (red, antibody labeling the presynaptic

    Article Snippet: Primary antibodies were mouse anti-CtBP2 (1:200, BD Biosciences), rabbit anti-GluA2/3 (1:200, Chemicon), rabbit anti-CaV 1.3 (1: 50, Alomone Labs), mouse anti-GluA2 (1:75 Chemicon), RIM2 (#140103 recognizing the PSD-95/discs large/ZO-1 (PDZ) domain, 1:200; Synaptic Systems), RIM1 (1:200; Synaptic Systems), RIM3 (1:100) , and Vglut3 (1:500; Synaptic Systems).

    Techniques: Immunohistochemistry, Mouse Assay, Immunolabeling, Antibody Labeling

    Disruption of RIM2 reduces synaptic Ca 2+ influx in IHCs. ( A ) Representative Ca 2+ currents of a RIM 1/2 cDKO (light gray), a RIM2α SKO IHC (dark gray), and a RIM2α con IHC (black) in response to 100-ms long-step depolarizations to −14

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

    Article Title: Rab3-interacting molecules 2α and 2β promote the abundance of voltage-gated CaV1.3 Ca2+ channels at hair cell active zones

    doi: 10.1073/pnas.1417207112

    Figure Lengend Snippet: Disruption of RIM2 reduces synaptic Ca 2+ influx in IHCs. ( A ) Representative Ca 2+ currents of a RIM 1/2 cDKO (light gray), a RIM2α SKO IHC (dark gray), and a RIM2α con IHC (black) in response to 100-ms long-step depolarizations to −14

    Article Snippet: Primary antibodies were mouse anti-CtBP2 (1:200, BD Biosciences), rabbit anti-GluA2/3 (1:200, Chemicon), rabbit anti-CaV 1.3 (1: 50, Alomone Labs), mouse anti-GluA2 (1:75 Chemicon), RIM2 (#140103 recognizing the PSD-95/discs large/ZO-1 (PDZ) domain, 1:200; Synaptic Systems), RIM1 (1:200; Synaptic Systems), RIM3 (1:100) , and Vglut3 (1:500; Synaptic Systems).

    Techniques: Immunohistochemistry, Mass Spectrometry

    The knockdown of spinal RIM1α expression relieves SNL-induced allodynia. A , B , Representative Western blot and statistical analysis (normalized to GAPDH) demonstrating that the intrathecal administration of RIM1α mRNA-targeting siRNA (RIM1α RNAi; 1, 3, and 5 μg; 10 μl; once daily for 4 d), but not missense siRNA (MS; 5 μg, 10 μl) or polyethylenimine (a transfection reagent, PEI; 10 μl), dose-dependently decreased spinal RIM1α expression of naive rat. Yet, all these treatments failed to affect the abundance of RIM2 in the dorsal horn sample of naive rat. it, Implantation of an intrathecal catheter; IB, immunoblotting. RIM1α, one-way ANOVA, post hoc Tukey's test, F (6,35) = 22.96, p

    Journal: The Journal of Neuroscience

    Article Title: Spinal Fbxo3-Dependent Fbxl2 Ubiquitination of Active Zone Protein RIM1α Mediates Neuropathic Allodynia through CaV2.2 Activation

    doi: 10.1523/JNEUROSCI.1732-16.2016

    Figure Lengend Snippet: The knockdown of spinal RIM1α expression relieves SNL-induced allodynia. A , B , Representative Western blot and statistical analysis (normalized to GAPDH) demonstrating that the intrathecal administration of RIM1α mRNA-targeting siRNA (RIM1α RNAi; 1, 3, and 5 μg; 10 μl; once daily for 4 d), but not missense siRNA (MS; 5 μg, 10 μl) or polyethylenimine (a transfection reagent, PEI; 10 μl), dose-dependently decreased spinal RIM1α expression of naive rat. Yet, all these treatments failed to affect the abundance of RIM2 in the dorsal horn sample of naive rat. it, Implantation of an intrathecal catheter; IB, immunoblotting. RIM1α, one-way ANOVA, post hoc Tukey's test, F (6,35) = 22.96, p

    Article Snippet: The sample was separated on an acrylamide gel and transferred to a polyvinylidene difluoride membrane, which was then incubated (1 h, room temperature) in either rabbit anti-RIM1α (1:250; Biorbyt), rabbit anti-RIM2 (1:400; GeneTex), goat anti-Fbxl2 antibody (1:250; Santa Cruz Biotechnology), rabbit anti-Fbxo3 antibody (1:500; Santa Cruz Biotechnology), mouse anti-ubiquitin antibody (1:1000; Santa Cruz Biotechnology), rabbit anti-CaV 2.2 (1:200; Thermo Fisher Scientific), N-cadherin (mouse; 1:2000; Thermo Fisher Scientific), or mouse anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH; 1:4000; Santa Cruz Biotechnology).

    Techniques: Expressing, Western Blot, Mass Spectrometry, Transfection

    Conditional knock-out of RIM2 in rod photoreceptors is under way as early as 3 weeks of age. A , Forms of RIM1 and RIM2 found in the brain and retina of wild-type and transgenic RIM1ff/2ff mice. Expression of RIM 1α ≫ 1β in the retina, which is similar to the proportions in whole-brain lysates (probed with Ms monoclonal anti-RIM1). The floxed RIM2ff ) and therefore all of the RIM2 in the transgenic animal is RIM2-cfp, which is revealed as an upward shift in its migration compared with brain lysates from wild-type animals (the cfp tag is nonfluorescent in the retina). Relative RIM2 expression levels in the brain are RIM 2α > 2β ∼ 2α-splice variants (α-sp.) and, in the retina, 2α and 2α-sp are equally expressed, but 2β is not detected. The regional distribution of RIM1/2 was examined by immunostaining cryosections of retina ∼10 μm thick ( B – F ). B , Expression of RIM1 is largely confined to the IPL and this is demonstrated with a rabbit polyclonal anti-RIM1 from Synaptic Systems (left) and a mouse monoclonal against RIM1 from BD Transduction Laboratories (right). Scale bar, 25 μm. C , D ), demarcating the OPL. The section in D is from a LMOPcre + transgenic animal, which shows an intensely stained patch of Cre in the ONL that is designated as a Cre region (samples in C and D are from RIM1ff/2ff littermates, 3 weeks of age). Bottom overview cropped to the OPL is labeled “all probes” because the signals are derived from anti-RIM2 staining (rabbit polyclonal, Alexa Fluor488, shown in green), the cone marker PNA (linked to Alexa Fluor 568, shown in red), and anti-CtBP2 (Ms monoclonal, Alexa Fluor 648, shown in blue). E , F , Regions selected for presentation at higher resolution. The control condition presented in C and E and the “Cre-poor region” in F show that the RIM2 signal overlaps extensively with CtBP2/Ribeye (aqua). The “Cre-rich region” in F reveals a drop in RIM2 staining. G , H ) and Synaptotagmin1 (Syt1, ∼60 kDa) were used as presynaptic markers, whereas CtBP2 (∼45 kDa) in the context of Western analysis represents the transcription factor that is expressed throughout the retina. I , Graphical summary of the densitometry measurements from Western analysis. Only RIM2 was reduced (see text for values) and no apparent change in the intensity of the synaptic markers was witnessed at 3 weeks. Data are presented as mean ± SE.

    Journal: The Journal of Neuroscience

    Article Title: RIM1/2-Mediated Facilitation of Cav1.4 Channel Opening Is Required for Ca2+-Stimulated Release in Mouse Rod Photoreceptors

    doi: 10.1523/JNEUROSCI.0658-15.2015

    Figure Lengend Snippet: Conditional knock-out of RIM2 in rod photoreceptors is under way as early as 3 weeks of age. A , Forms of RIM1 and RIM2 found in the brain and retina of wild-type and transgenic RIM1ff/2ff mice. Expression of RIM 1α ≫ 1β in the retina, which is similar to the proportions in whole-brain lysates (probed with Ms monoclonal anti-RIM1). The floxed RIM2ff ) and therefore all of the RIM2 in the transgenic animal is RIM2-cfp, which is revealed as an upward shift in its migration compared with brain lysates from wild-type animals (the cfp tag is nonfluorescent in the retina). Relative RIM2 expression levels in the brain are RIM 2α > 2β ∼ 2α-splice variants (α-sp.) and, in the retina, 2α and 2α-sp are equally expressed, but 2β is not detected. The regional distribution of RIM1/2 was examined by immunostaining cryosections of retina ∼10 μm thick ( B – F ). B , Expression of RIM1 is largely confined to the IPL and this is demonstrated with a rabbit polyclonal anti-RIM1 from Synaptic Systems (left) and a mouse monoclonal against RIM1 from BD Transduction Laboratories (right). Scale bar, 25 μm. C , D ), demarcating the OPL. The section in D is from a LMOPcre + transgenic animal, which shows an intensely stained patch of Cre in the ONL that is designated as a Cre region (samples in C and D are from RIM1ff/2ff littermates, 3 weeks of age). Bottom overview cropped to the OPL is labeled “all probes” because the signals are derived from anti-RIM2 staining (rabbit polyclonal, Alexa Fluor488, shown in green), the cone marker PNA (linked to Alexa Fluor 568, shown in red), and anti-CtBP2 (Ms monoclonal, Alexa Fluor 648, shown in blue). E , F , Regions selected for presentation at higher resolution. The control condition presented in C and E and the “Cre-poor region” in F show that the RIM2 signal overlaps extensively with CtBP2/Ribeye (aqua). The “Cre-rich region” in F reveals a drop in RIM2 staining. G , H ) and Synaptotagmin1 (Syt1, ∼60 kDa) were used as presynaptic markers, whereas CtBP2 (∼45 kDa) in the context of Western analysis represents the transcription factor that is expressed throughout the retina. I , Graphical summary of the densitometry measurements from Western analysis. Only RIM2 was reduced (see text for values) and no apparent change in the intensity of the synaptic markers was witnessed at 3 weeks. Data are presented as mean ± SE.

    Article Snippet: Other antibodies used were as follows: anti-RIM2, rabbit polyclonal (Synaptic Systems, 140–303); anti-RIM1, rabbit polyclonal (Synaptic Systems, 140–003) and anti-RIM1 mouse monoclonal (BD Transduction Laboratories, 610907); anti-Fodrin/α-spectrin, clone AA6, mouse monoclonal (ICN Biomedicals); lectin PNA-Alexa Fluor 568 (Invitrogen, ): anti-Ribeye/U2656 rabbit polyclonal antibody ( ); anti-CtBP2 mouse monoclonal (BD Transduction Laboratories, 612044); anti-Cre Recombinase, rabbit polyclonal (Novagen, 69050); and anti-Cre recombinase, mouse monoclonal (Covance, MMS-106P).

    Techniques: Knock-Out, Transgenic Assay, Mouse Assay, Expressing, Mass Spectrometry, Migration, Immunostaining, Transduction, Staining, Labeling, Derivative Assay, Marker, Western Blot

    RIM2 is selectively removed from rods expressing Cre by 8 weeks and persists at cone terminals. A , Colabeling of cryosections from 8-week-old littermates. The retinal section from a control ( RIM1ff/2ff ) on the left has a continuous line of RIM2 staining across the OPL (shown in green, and transmitted light is overlaid in blue to demarcate the ONL's rod somata); in contrast, the section from the cdko animal presented on the right has two regions of Cre staining (pink) and the underlying OPL lacks significant RIM2. B – D , Additional probes were used to demonstrate the absence of RIM2 at rod ribbons in regions enriched with Cre, and the constancy of RIM2 at cone terminals. B , C , Top, black and white images are composites of transmitted light and fluorescent signals from anti-CtBP2 and anti-Cre immunolabeling (both are Ms monoclonal antibodies and visualized with anti-Ms IgG-Alexa Fluor 648; see Materials and Methods). The transmitted light images outline the ONL and anti-CtBP2 stains Ribeye/ribbons demarcates the OPL. The lower color images in B and C show immunostaining for the antigens indicated (anti-RIM2, rabbit polyclonal, Alexa Fluor 488, shown in green; the cone marker PNA linked to Alexa Fluor 568, shown in red; and anti-CtBP2, Ms monoclonal, Alexa Fluor 648, shown in blue), and the overlay of the various probes are presented in the bottom panels. The control in B exhibits a continuous band of RIM2 labeling in the OPL, whereas the cdko retina shown in C expresses a patch of Cre that is accompanied by a significant disruption in RIM2 in the underlying OPL. D , RIM2 staining that remains in Cre regions is localized to “cone terminals” that are labeled with PNA and occupy the innermost aspects of the OPL. The regions where the rod terminals are located are indicated with turquoise arrowheads. The control exhibits RIM2, which is seen to colocalize with CtBP2 in the overlay. In contrast, RIM2-free rod ribbons are seen in the Cre + region, yet the underlying cone expresses RIM2, which is a consequence of the cones not expressing Cre. E , RIM2 is heavily expressed in the OPL and a major portion of RIM2 is expressed in the IPL. F , Example of a Western blot showing the decreased levels of RIM2 in cdko animals (3 months of age). G , Summary plots of RIM2 and other synaptic protein levels found in retinas of 3-month-old mice. Data are presented as mean ± SE.

    Journal: The Journal of Neuroscience

    Article Title: RIM1/2-Mediated Facilitation of Cav1.4 Channel Opening Is Required for Ca2+-Stimulated Release in Mouse Rod Photoreceptors

    doi: 10.1523/JNEUROSCI.0658-15.2015

    Figure Lengend Snippet: RIM2 is selectively removed from rods expressing Cre by 8 weeks and persists at cone terminals. A , Colabeling of cryosections from 8-week-old littermates. The retinal section from a control ( RIM1ff/2ff ) on the left has a continuous line of RIM2 staining across the OPL (shown in green, and transmitted light is overlaid in blue to demarcate the ONL's rod somata); in contrast, the section from the cdko animal presented on the right has two regions of Cre staining (pink) and the underlying OPL lacks significant RIM2. B – D , Additional probes were used to demonstrate the absence of RIM2 at rod ribbons in regions enriched with Cre, and the constancy of RIM2 at cone terminals. B , C , Top, black and white images are composites of transmitted light and fluorescent signals from anti-CtBP2 and anti-Cre immunolabeling (both are Ms monoclonal antibodies and visualized with anti-Ms IgG-Alexa Fluor 648; see Materials and Methods). The transmitted light images outline the ONL and anti-CtBP2 stains Ribeye/ribbons demarcates the OPL. The lower color images in B and C show immunostaining for the antigens indicated (anti-RIM2, rabbit polyclonal, Alexa Fluor 488, shown in green; the cone marker PNA linked to Alexa Fluor 568, shown in red; and anti-CtBP2, Ms monoclonal, Alexa Fluor 648, shown in blue), and the overlay of the various probes are presented in the bottom panels. The control in B exhibits a continuous band of RIM2 labeling in the OPL, whereas the cdko retina shown in C expresses a patch of Cre that is accompanied by a significant disruption in RIM2 in the underlying OPL. D , RIM2 staining that remains in Cre regions is localized to “cone terminals” that are labeled with PNA and occupy the innermost aspects of the OPL. The regions where the rod terminals are located are indicated with turquoise arrowheads. The control exhibits RIM2, which is seen to colocalize with CtBP2 in the overlay. In contrast, RIM2-free rod ribbons are seen in the Cre + region, yet the underlying cone expresses RIM2, which is a consequence of the cones not expressing Cre. E , RIM2 is heavily expressed in the OPL and a major portion of RIM2 is expressed in the IPL. F , Example of a Western blot showing the decreased levels of RIM2 in cdko animals (3 months of age). G , Summary plots of RIM2 and other synaptic protein levels found in retinas of 3-month-old mice. Data are presented as mean ± SE.

    Article Snippet: Other antibodies used were as follows: anti-RIM2, rabbit polyclonal (Synaptic Systems, 140–303); anti-RIM1, rabbit polyclonal (Synaptic Systems, 140–003) and anti-RIM1 mouse monoclonal (BD Transduction Laboratories, 610907); anti-Fodrin/α-spectrin, clone AA6, mouse monoclonal (ICN Biomedicals); lectin PNA-Alexa Fluor 568 (Invitrogen, ): anti-Ribeye/U2656 rabbit polyclonal antibody ( ); anti-CtBP2 mouse monoclonal (BD Transduction Laboratories, 612044); anti-Cre Recombinase, rabbit polyclonal (Novagen, 69050); and anti-Cre recombinase, mouse monoclonal (Covance, MMS-106P).

    Techniques: Expressing, Staining, Immunolabeling, Mass Spectrometry, Immunostaining, Marker, Labeling, Western Blot, Mouse Assay