Imaging artifacts from animal motion could induce transients in the fluorescence signals. This issue is particularly relevant for small neuronal compartments such as dendritic spines and axonal boutons. To characterize motion-related signals in our setup, we injected AAV1-CMVIVS-DIO-EGFP (Vector Biolabs; 6 × 1013 GC/mL titer, 184 nL) into Cg1/M2 in adult male CamKIIa-Cre [37 (link)] (Stock No. 005359, Jackson Laboratory) and SST-IRES-Cre animals to image apical dendritic spines of excitatory neurons (n = 76 spines, 3 animals) and SST axonal boutons (n = 72 boutons, 3 animals) respectively. Imaging conditions were the same as our calcium imaging experiments. Analyses were the same, except there was no regression and subtraction of shaft signals for dendritic spines, and no clustering for axonal boutons. We compared the results of these control EGFP experiments to the GCaMP6f experiments to quantify the extent of motion artifacts in our data.
Camkiia cre
Manufactured by Jackson ImmunoResearch
CamKIIa-Cre is a recombinant Cre recombinase enzyme expressed under the control of the calcium/calmodulin-dependent protein kinase IIa (CaMKIIa) promoter. The CaMKIIa promoter provides expression in excitatory neurons in the forebrain.
Automatically generated - may contain errors
4 protocols using camkiia cre
1
Characterizing Motion Artifacts in Neuronal Imaging
2
Optogenetic Modulation of Hippocampal Neurons
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All animals were bred at the local animal facility and kept in 12-h light/dark cycle with access to food and water ad libitum. All procedures were approved by the Malmö/Lund Animal Research Ethics Board, permit M47-15 and M49-15.
To explore the possibility of generating epileptiform afterdischarges by selective activation of interneurons or excitatory neurons in hippocampal slices we used 2 separate mouse lines. The Ai32(RCL-ChR2(H134R)/EYFP; Jackson #012569) mouse line was bred with either a PV-Cre (Jackson #008069) or a CaMKIIa-Cre (Jackson #005359) mice to obtain off-spring expressing ChR2 under the PV-promoter (PV-ChR2 mice) or the CaMKII-alpha promoter (CamKIIa-ChR2 mice).
To explore the possibility of generating epileptiform afterdischarges by selective activation of interneurons or excitatory neurons in hippocampal slices we used 2 separate mouse lines. The Ai32(RCL-ChR2(H134R)/EYFP; Jackson #012569) mouse line was bred with either a PV-Cre (Jackson #008069) or a CaMKIIa-Cre (Jackson #005359) mice to obtain off-spring expressing ChR2 under the PV-promoter (PV-ChR2 mice) or the CaMKII-alpha promoter (CamKIIa-ChR2 mice).
3
Characterizing Motion Artifacts in Neuronal Imaging
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Imaging artifacts from animal motion could induce transients in the fluorescence signals. This issue is particularly relevant for small neuronal compartments such as dendritic spines and axonal boutons. To characterize motion-related signals in our setup, we injected AAV1-CMVIVS-DIO-EGFP (Vector Biolabs; 6 × 1013 GC/mL titer, 184 nL) into Cg1/M2 in adult male CamKIIa-Cre [37 (link)] (Stock No. 005359, Jackson Laboratory) and SST-IRES-Cre animals to image apical dendritic spines of excitatory neurons (n = 76 spines, 3 animals) and SST axonal boutons (n = 72 boutons, 3 animals) respectively. Imaging conditions were the same as our calcium imaging experiments. Analyses were the same, except there was no regression and subtraction of shaft signals for dendritic spines, and no clustering for axonal boutons. We compared the results of these control EGFP experiments to the GCaMP6f experiments to quantify the extent of motion artifacts in our data.
4
Transgenic Mouse Lines for Neural Circuit Analysis
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All housing of animals and procedures were approved by the University of Michigan Institutional Animal Care and Use Committee. Multiple mouse lines were used in this study, including PV-IRES-Cre (Jackson Laboratories, 008069), CaMKIIa-Cre (Jackson Laboratories, 005359), Ai32 (Jackson Laboratories, 024109), Ai14 (Jackson Laboratories, 007914), PV-IRES-Cre x Ai14 (crossed in house), PV-IRES-Cre x Ai32 (crossed in house), CaMKIIa-Cre x Ai32 (crossed in house), and NTSR1-Cre (MMRRC, 030648-UCD).
All mice excluding the NTSR1-Cre line were on a C57Bl6 background, while the NTSR1-Cre mice had a mixed C57Bl6/ICR background. Mice of both sexes between the ages of P21-31 and P60-65 were included in the experiments.
All mice excluding the NTSR1-Cre line were on a C57Bl6 background, while the NTSR1-Cre mice had a mixed C57Bl6/ICR background. Mice of both sexes between the ages of P21-31 and P60-65 were included in the experiments.
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