recording nmda receptor mediated epscs  (Tocris)

Journal: Advanced Science

Article Title: Cell‐Type Specific Circuits in the Mammillary Body for Place and Object Recognition Memory

doi: 10.1002/advs.202409397

Figure Lengend Snippet: Electrophysiological properties of Kcnn4 and Cacna1h in PV and Drd2 neurons. A) A strategy for rAAV2/9‐mediated CRISPR‐Cas9 in vivo gene knockdown. B) Representative images showing the expression of Kcnn4 and Cacna1h sgRNAs in PV (red) and Drd2 (Green) neurons. C) Representative traces and a bar graph showing SAPs in PV neurons with (PV Kcnn4− ) or without (PV Kcnn4+ ) Kcnn4 knockdown, sgRNAs target to zfy2 as the negative control (PV zfy2− ). Data are mean ± SEM ( n = 18 neurons per group, adjusted **** p < 0.0001, one‐way ANOVA followed with Bonferroni's post hoc test). D) Representative traces and a bar graph showing RBs in Drd2 neurons with (Drd2 Cacan1h− ) and without (Drd2 Cacan1h+ ) Cacan1h knockdown, sgRNAs target to zfy2 as the negative control (Drd2 zfy2− ). Data are mean ± SEM ( n = 18 neurons per group, adjusted **** P < 0.0001, one‐way ANOVA followed by Bonferroni's post hoc test). E) Representative traces and bar graphs showing spontaneous miniature EPSCs in PV and Drd2 neurons with (PV Kcnn4− and Drd2 Cacna1h− ) or without (PV Kcnn4+ and Drd2 Cacna1h+ ) Kcnn4 or Cacna1h knockdown. Data are mean ± SEM ( n = 18 neurons per group, t ‐test).

Article Snippet: For recording NMDA receptor‐mediated EPSCs, membrane potentials were hold at +60 mV in the presence of 20 × 10 −6 m bicuculline (BIC, TOCRIS, 0130) and 20 × 10 −6 m CNQX (TOCRIS, 0373).

Techniques: CRISPR, In Vivo, Knockdown, Expressing, Negative Control

Journal: Advanced Science

Article Title: Cell‐Type Specific Circuits in the Mammillary Body for Place and Object Recognition Memory

doi: 10.1002/advs.202409397

Figure Lengend Snippet: Structurally and functionally distinct cell‐type specific subcircuits. A–C) Representative images (A and B) showing GFP (green) in PV TVA/G neurons (A) and their presynaptic pyramidal neurons (B) in the dorsal subiculum (DS). A bar graph (C) showing the numbers of GFP‐expressing neurons in each of these brain regions. Data are mean ± SEM ( n = 5 mice per group, adjusted *** P < 0.001, **** P < 0.0001, one‐way ANOVA followed with Bonferroni's post hoc test). HMI: high‐magnification image showing single DS neuron. D–F) Representative images (D and E) showing tdT (red) in Drd2 TVA/G neurons (D) and their presynaptic pyramidal neurons (E) in the ventral subiculum (VS). A bar graph (F) showing the numbers of tdT‐expressing neurons in each of these brain regions. Data are mean ± SEM ( n = 5 mice per group, adjusted **** P < 0.0001, one‐way ANOVA followed with Bonferroni's post hoc test). HMI: high‐magnification image showing single VS neuron. G,H) Experimental designs and whole‐cell patch‐clamp recordings from PV (G) and Drd2 (H) neurons. Representative traces of NMDA receptor‐mediated postsynaptic currents (EPSCs) at a holding potential of +60 mV were evoked by stimulating DS ChR2 (DS→PV, blue) or VS ChR2 (VS→Drd2, green) terminals in the presence of 20 × 10 −6 m CNQX. EPSCs were blocked by 1 × 10 −6 m TTX and rescued by 100 × 10 −6 m 4‐AP and were sensitive to 50 × 10 −6 m AP‐5. OGS: Optogenetic stimulation. Bar graphs showing the mean amplitudes of EPSCs. Data are mean ± SEM ( n = 15 neurons per group, adjusted **** P < 0.0001, one‐way ANOVA followed with Bonferroni's post hoc test).

Article Snippet: For recording NMDA receptor‐mediated EPSCs, membrane potentials were hold at +60 mV in the presence of 20 × 10 −6 m bicuculline (BIC, TOCRIS, 0130) and 20 × 10 −6 m CNQX (TOCRIS, 0373).

Techniques: Expressing, Patch Clamp

Journal: The Journal of Headache and Pain

Article Title: Alleviation of migraine related pain and anxiety by inhibiting calcium-stimulating AC1-dependent CGRP in the insula of adult rats

doi: 10.1186/s10194-024-01778-3

Figure Lengend Snippet: Enhanced NMDA receptors-mediated currents and their expression in the IC. A The synaptic input-output curve in slices from IS rats was steeper than in those from PBS group (IS: n =10 neurons/6 rats, PBS: n =11 neurons/5 rats; two-way ANOVA with Bonferroni post hoc). B I-V curves of NMDAR-EPSCs of IC pyramidal neurons recorded at holding potentials ranging from − 80 to + 40 mV in PBS and IS rats (IS: n =12 neurons/5 rats, PBS: n =10 neurons/5 rats; two-way ANOVA with Bonferroni post hoc). C A timeline plot of one representative neuron in IC showing the contribution of NMDA GluN2A and GluN2B -mediated currents (left). PEAQX (0.4 μM): selective GluN2A antagonist, Ro25-6981 (3 μM): selective GluN2B antagonist. Summarized data showing the contribution of GluN2A (two-tailed independent sample t-test) and GluN2B (two-tailed independent sample rank sum test) -mediated currents in IC neurons (IS: n =9 neurons/6 rats, PBS: n =6 neurons/5 rats; right). D Representative western blotting for GluN2B and p-GluN2B-S1303 in the IC from IS and PBS rats. E The total protein levels of GluN2B, p-GluN2B-S1303 significantly enhanced in IS rats ( n =5 rats/group; two-tailed independent sample t-test). F The phosphorylation level of GluN2B, p-GluN2B-S1303, significantly enhanced in IS rats ( n =5 rats/group; two-tailed independent sample t-test). All data are presented as the mean ± SEM (# P <0.05, ## P <0.01, IS vs. PBS)

Article Snippet: NMDA receptor-mediated EPSCs were recorded at 30 mV by bathing with CNQX (20 mM, Tocris, UK).

Techniques: Expressing, Two Tailed Test, Western Blot, Phospho-proteomics