Journal: Neuron

Article Title: A Photoactivatable Botulinum Neurotoxin for Inducible Control of Neurotransmission

doi: 10.1016/j.neuron.2019.01.002

Figure Lengend Snippet: (A) Postsynaptic Ca2+ transients arising from quantal neurotransmitter release can be detected with jRGECO1a. Top: dendritic segment from a cultured hippocampal neuron expressing jRGECO1a. Center: jRGECO1a within a single dendritic spine before, during, and 2 s after a spontaneous Ca2+ transient. Bottom: kymograph generated from the red line drawn through the spine (sp) and shaft (sh) at the top. Two discrete events (arrowheads) can be observed in this example.

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Monoclonal anti-VAMP2 Synaptic Systems Cat# 104 211BT; RRID: AB_2619758 Polyclonal anti-mCh Thermo Fisher Scientific Cat# PA5-34974; RRID: AB_2552323 Goat anti-rabbit IRDye 800CW secondary Li-Cor Cat# 926-32211; RRID: AB_621833 Polyclonal anti-GFP Sigma-Aldrich Cat# G1544; RRID: AB_439690 Polyclonal anti-GFP Thermo/Molecular Probes Cat#A-11122; RRID: AB_221569 Goat anti-mouse Alexa647 conjugated secondary antibody Life Technologies/Thermo Fisher Cat#A-32728; RRID: AB_2633277 Chemicals, Peptides, and Recombinant Proteins FM4-64 Molecular Probes/Thermo Fisher Cat #13320 FM1-43FX (fixable version of FM1-43) Molecular Probes/Thermo Fisher Cat # {"type":"entrez-nucleotide","attrs":{"text":"F35355","term_id":"4820981","term_text":"F35355"}} F35355 Experimental Models: Cell Lines HEK293T ATCC RRID: CVCL_0063 Experimental Models: Organisms/Strains Rat, Sprague Dawley Charles River Crl:CD(SD)BR; RRID: RGD_734476 C. elegans wild type Bristol N2 CGC RRID: WB-STRAIN:N2_ancestral Mouse, C57BL/6 The Jackson Laboratory JAX:000664; RRID: IMSR_JAX:000664 Recombinant DNA jRGECO1a Dana et al., 2016 Addgene #61563 Cryptochrome2 Kennedy et al., 2010 Addgene #26866 CIBN Kennedy et al., 2010 Addgene #26867 SspBmicro Guntas et al., 2015 Addgene #60416 iLID Guntas et al., 2015 Addgene #60413 SspBwt/nano Guntas et al., 2015 Addgene #60415 pEGFP-VAMP3 Galli et al., 1998 Addgene #42310 BoNT/B LC (S. cerevisiae codon optimized) This manuscript N/A BoNT/B LC (M. musculus codon optimized) This manuscript N/A Software and Algorithms MATLAB MathWorks https://www.mathworks.com/ Igor Pro neuromatic package Wavemetrics http://www.neuromatic.thinkrandom.com Mini analysis Synaptosoft http://www.synaptosoft.com Andor IQ3 acquisition software package Andor Technologies https://andor.oxinst.com/products/iq-live-cell-imaging-software Metamorph Molecular Devices https://www.moleculardevices.com/products/cellular-imaging-systems/acquisition-and-analysis-software/metamorph-microscopy ImageJ National Institutes of Health https://imagej.nih.gov/ij/ WinLTP University of Bristol https://www.winltp.com/ Open in a separate window KEY RESOURCES TABLE Few tools allow inhibition of neural activity for long time periods with light We engineered botulinum neurotoxin B so that it can be activated with blue light Photoactivated botulinum toxin efficiently cleaves the SNARE protein VAMP2 We demonstrate utility in diverse systems, from mammalian brain slices to worms

Techniques: Cell Culture, Expressing, Generated

Journal: Neuron

Article Title: A Photoactivatable Botulinum Neurotoxin for Inducible Control of Neurotransmission

doi: 10.1016/j.neuron.2019.01.002

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Monoclonal anti-VAMP2 Synaptic Systems Cat# 104 211BT; RRID: AB_2619758 Polyclonal anti-mCh Thermo Fisher Scientific Cat# PA5-34974; RRID: AB_2552323 Goat anti-rabbit IRDye 800CW secondary Li-Cor Cat# 926-32211; RRID: AB_621833 Polyclonal anti-GFP Sigma-Aldrich Cat# G1544; RRID: AB_439690 Polyclonal anti-GFP Thermo/Molecular Probes Cat#A-11122; RRID: AB_221569 Goat anti-mouse Alexa647 conjugated secondary antibody Life Technologies/Thermo Fisher Cat#A-32728; RRID: AB_2633277 Chemicals, Peptides, and Recombinant Proteins FM4-64 Molecular Probes/Thermo Fisher Cat #13320 FM1-43FX (fixable version of FM1-43) Molecular Probes/Thermo Fisher Cat # {"type":"entrez-nucleotide","attrs":{"text":"F35355","term_id":"4820981","term_text":"F35355"}} F35355 Experimental Models: Cell Lines HEK293T ATCC RRID: CVCL_0063 Experimental Models: Organisms/Strains Rat, Sprague Dawley Charles River Crl:CD(SD)BR; RRID: RGD_734476 C. elegans wild type Bristol N2 CGC RRID: WB-STRAIN:N2_ancestral Mouse, C57BL/6 The Jackson Laboratory JAX:000664; RRID: IMSR_JAX:000664 Recombinant DNA jRGECO1a Dana et al., 2016 Addgene #61563 Cryptochrome2 Kennedy et al., 2010 Addgene #26866 CIBN Kennedy et al., 2010 Addgene #26867 SspBmicro Guntas et al., 2015 Addgene #60416 iLID Guntas et al., 2015 Addgene #60413 SspBwt/nano Guntas et al., 2015 Addgene #60415 pEGFP-VAMP3 Galli et al., 1998 Addgene #42310 BoNT/B LC (S. cerevisiae codon optimized) This manuscript N/A BoNT/B LC (M. musculus codon optimized) This manuscript N/A Software and Algorithms MATLAB MathWorks https://www.mathworks.com/ Igor Pro neuromatic package Wavemetrics http://www.neuromatic.thinkrandom.com Mini analysis Synaptosoft http://www.synaptosoft.com Andor IQ3 acquisition software package Andor Technologies https://andor.oxinst.com/products/iq-live-cell-imaging-software Metamorph Molecular Devices https://www.moleculardevices.com/products/cellular-imaging-systems/acquisition-and-analysis-software/metamorph-microscopy ImageJ National Institutes of Health https://imagej.nih.gov/ij/ WinLTP University of Bristol https://www.winltp.com/ Open in a separate window KEY RESOURCES TABLE Few tools allow inhibition of neural activity for long time periods with light We engineered botulinum neurotoxin B so that it can be activated with blue light Photoactivated botulinum toxin efficiently cleaves the SNARE protein VAMP2 We demonstrate utility in diverse systems, from mammalian brain slices to worms

Techniques: Recombinant, Software