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

Article Title: CaMKII modulates sodium current in neurons from epileptic Scn2a mutant mice

doi: 10.1073/pnas.1615774114

Figure Lengend Snippet: Biophysical properties of Scn2a Q54 neuronal sodium channels

Article Snippet: Using two separate assays that are specific for the transgene, we found no difference in hippocampal transgene levels between B6.Q54 and F1.Q54 animals ( ; Assay Rn00680558: B6.Q54, 0.19 ± 0.03 vs. F1.Q54, 0.22 ± 0.05; Assay Rn00561862: B6.Q54, 0.13 ± 0.02 vs. F1.Q54 0.15 ± 0.05), thereby excluding divergent expression of the transgene.

Techniques:

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

Article Title: CaMKII modulates sodium current in neurons from epileptic Scn2a mutant mice

doi: 10.1073/pnas.1615774114

Figure Lengend Snippet: Analysis of transgene expression level. Quantification of transgene transcript expression was determined by ddPCR of hippocampal first-strand cDNA using two separate assays specific for rat Scn2a. Relative transcript levels are expressed as a ratio of rat Scn2a concentration to Tbp concentration. Data are presented as mean ± SEM of five to seven biological replicates.

Article Snippet: Using two separate assays that are specific for the transgene, we found no difference in hippocampal transgene levels between B6.Q54 and F1.Q54 animals ( ; Assay Rn00680558: B6.Q54, 0.19 ± 0.03 vs. F1.Q54, 0.22 ± 0.05; Assay Rn00561862: B6.Q54, 0.13 ± 0.02 vs. F1.Q54 0.15 ± 0.05), thereby excluding divergent expression of the transgene.

Techniques: Expressing, Concentration Assay

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

Article Title: CaMKII modulates sodium current in neurons from epileptic Scn2a mutant mice

doi: 10.1073/pnas.1615774114

Figure Lengend Snippet: Effect of intracellular Ca2+ on neuronal sodium current. (A) Representative normalized current trace showing persistent sodium current in response to a 200-ms depolarization (to 0 mV) from HEK293T cells transfected with NaV1.2-Q54 in the presence of 0 μM Ca2+ (black trace) or 1 μM Ca2+ (red trace) neurons. Inset shows final 50 ms of the 200-ms depolarized voltage step. (B) Quantification of persistent current levels of NaV1.2-Q54 in the presence of 0 μM Ca2+ or 1 μM Ca2+ expressed as percent of peak current. (C) Steady-state inactivation for sodium channels in the presence of 0 μM Ca2+ (black symbols) or 1 μM Ca2+ (red symbols). All data are expressed as mean ± SEM for n = 9–11 cells (*P < 0.05).

Article Snippet: Using two separate assays that are specific for the transgene, we found no difference in hippocampal transgene levels between B6.Q54 and F1.Q54 animals ( ; Assay Rn00680558: B6.Q54, 0.19 ± 0.03 vs. F1.Q54, 0.22 ± 0.05; Assay Rn00561862: B6.Q54, 0.13 ± 0.02 vs. F1.Q54 0.15 ± 0.05), thereby excluding divergent expression of the transgene.

Techniques: Transfection

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

Article Title: CaMKII modulates sodium current in neurons from epileptic Scn2a mutant mice

doi: 10.1073/pnas.1615774114

Figure Lengend Snippet: CaMKII regulation of voltage-gated sodium current. (A and B, Left) Representative normalized current trace illustrating persistent sodium current in response to a 200-ms depolarization (to 0 mV) recorded from HEK293T cells transfected with NaV1.2-Q54 (A) or NaV1.2-WT (B) in the presence of activated CaMKII monomer (blue trace) or vehicle (black trace). (A and B, Center) Quantification of persistent current levels of NaV1.2-Q54 (A) or NaV1.2-WT (B) in the presence of activated CaMKII monomer or vehicle expressed as percent of peak current. (A and B, Right) Steady-state inactivation for sodium channels in the presence of activated CaMKII monomer (blue symbols) or vehicle (black symbols). (C, Left) Representative normalized current trace illustrating persistent sodium current in response to a 200-ms depolarization (to 0 mV) recorded from F1.Q54 pyramidal neurons in the presence of 10 μM KN-93 (black trace) or 10 μM KN-92 (blue trace). (C, Center) Quantification of persistent current levels from pyramidal neurons in the presence of 10 μM KN-93 or 10 μM KN-92 expressed as percent of peak current. (C, Right) Steady-state inactivation for sodium channels in the presence of 10 μM KN-93 (black symbols) or 10 μM KN-92 (blue symbols). Open symbols represent individual cells, whereas closed symbols are expressed as mean ± SEM for n = 3–7 cells (*P < 0.05).

Article Snippet: Using two separate assays that are specific for the transgene, we found no difference in hippocampal transgene levels between B6.Q54 and F1.Q54 animals ( ; Assay Rn00680558: B6.Q54, 0.19 ± 0.03 vs. F1.Q54, 0.22 ± 0.05; Assay Rn00561862: B6.Q54, 0.13 ± 0.02 vs. F1.Q54 0.15 ± 0.05), thereby excluding divergent expression of the transgene.

Techniques: Transfection