Journal: STAR Protocols

Article Title: Protocol for whole-cell patch-clamp recording and post hoc identification of hippocampal CA2 pyramidal neurons in adult mouse brain slices

doi: 10.1016/j.xpro.2026.104470

Figure Lengend Snippet: Passive properties of CA2 pyramidal neurons and miniature IPSCs (A) Capacitive and leakage currents in a CA2 pyramidal neuron in response to a 5 mV voltage step recorded at a holding potential of Vh=−70 mV before the current trace in A. The series resistance Rs is 8.1 MΩ. Capacitive transients are fitted with the sum of two exponentials giving time constants of 30 and 355 pF. Read out value from clampEx was Cm=210 pF. Current trace is an average of 9 raw traces. (B) Capacitive and leakage currents acquired after the current trace in C to show the absence of gross change in the passive properties of the cell and recording conditions. Rs is 8.8 MΩ (less than 9% change in comparison to condition in A). Average current trace from 11 raw traces. Current traces in A and B are recorded in the presence of 20 μM D-AP5, 10 μM CNQX, and 1 μM TTX.

Article Snippet: D-AP5 , Hello Bio , Cat#HB0225.

Techniques: Comparison

Journal: STAR Protocols

Article Title: Protocol for whole-cell patch-clamp recording and post hoc identification of hippocampal CA2 pyramidal neurons in adult mouse brain slices

doi: 10.1016/j.xpro.2026.104470

Figure Lengend Snippet: Excitatory and inhibitory spontaneous synaptic currents in CA2 pyramidal neurons (A) Representative recording of mIPSCs at a holding potential of Vh=−70 mV. 20 μM D-AP5, 10 μM CNQX, and 1 μM TTX are added to external solution to block excitatory synaptic activity and spontaneous AP firing. Green dashed vertical lines indicate the sections of current selected for expanded time view in B. Current trace sections are labeled #1 to #5. (B) Selection of five current trace sections extracted from current trace in (C). (C) Miniature currents selected using Clampfit with template matching selection method. Superimposition of 23 individual current traces in black with one in red. (D) Current traces from (C) at expanded time scale. (E) Original trace of sEPSCs recorded at a holding potential of Vh=−70 mV. 50 μM picrotoxin was added to external solution to block inhibitory synaptic activity. Blue dashed vertical lines indicate the sections of current selected for expanded time view in F. (F) Five current trace segments (blue label #1 to #5) extracted from the whole current trace in (E) at expanded time scale. (G) Spontaneous excitatory postsynaptic currents selected using Clampfit with template matching selection method. Superimposition of 12 individual current traces in black with one in red. (H) Current traces from (G) at expanded time scale.

Article Snippet: D-AP5 , Hello Bio , Cat#HB0225.

Techniques: Blocking Assay, Activity Assay, Labeling, Selection

Journal: Advanced Science

Article Title: Fine‐Tuned Regulation of mRNA Translation and Transport by STAU2 Condensate Facilitates Neuronal Development and Plasticity

doi: 10.1002/advs.202600044

Figure Lengend Snippet: Synaptic activity controls local translation of RGS4 mRNA in a STAU2‐dependent manner. (A) Representative confocal images of DIV14 rat hippocampal neurons showing endogenous STAU2 (red) and the postsynaptic marker Homer1c (cyan) under three conditions: untreated (“resting”), excitatory (“+Bic”), or silenced (“+TTX”). Scale bars, 2 µm (left) and 1 µm (right). (B) Quantification of the percentage of endogenous STAU2 localized to dendritic spines. Data are mean ± SEM (Rest, n = 50; +Bic, n = 48; +TTX, n = 36), ** p < 0.01, *** p < 0.001, one‐way ANOVA. (C) Schematic of the RGS4 SunTag reporter. Nascent RGS4 polypeptides carry tandem SunTag epitopes recognized by co‐expressed ScFv‐GFP (top). STAU2 overexpression (STAU2‐OE) or knockdown (siSTAU2) modulates the translation complex (bottom). (D) Representative confocal images of dendrites from DIV14 rat hippocampal neurons co‐expressing the SunTag reporter with either empty vector (Control), mCherry‐STAU2 (STAU2‐OE), or siRNA against STAU2 (siSTAU2) under the three synaptic conditions described above. Newly synthesized RGS4 peptides detected by ScFv‐GFP in dendritic spines are indicated by arrowheads. Scale bars, 5 µm. (E) Quantification of relative SunTag/ScFv‐GFP intensity in dendritic spines under the conditions shown in D. Values are normalized to the resting Control group. Data are mean ± SEM ( n = 65, 64, 72, 33, 32, 34, 34, 32, 32 from left to right), *** p < 0.001, one‐way ANOVA. (F) Fold change in nascent RGS4 synthesis in dendritic spines induced by excitatory (“+Bic”) or silencing (“+TTX”) treatments relative to resting conditions in Control, STAU2‐OE, and siSTAU2 neurons. Data are mean ± SEM (same n values as in E), * p < 0.05, ** p < 0.01, one‐way ANOVA. (G) Model summarizing how synaptic activity bidirectionally regulates STAU2–RNP assembly and local RGS4 translation at synapses. Under silenced conditions (“+TTX”, left), STAU2 forms large condensates at postsynaptic sites and strongly represses RGS4 translation. At rest (middle), STAU2 forms baseline condensates that allow appropriate levels of RGS4 synthesis. Under activated conditions (“+Bic,” right), STAU2 condensates disperse into smaller complexes, relieving repression and promoting robust production of nascent RGS4 peptides in dendritic spines.

Article Snippet: For silencing conditions (“+TTX”), neurons were treated for 4 h with 1 μM TTX (Shanghai Charm‐Analysis, 20–309500), 50 μM D‐AP5 (MCE, HY‐100714A), and 100 μM DNQX (Abcam, ab120018) [ ].

Techniques: Activity Assay, Marker, Over Expression, Knockdown, Expressing, Plasmid Preparation, Control, Synthesized

Journal: Endocrinology

Article Title: Robust serotonin activation of the kisspeptin GnRH pulse generator in male and female mice

doi: 10.1210/endocr/bqag034

Figure Lengend Snippet: Effects of serotonin and 5-HT receptor compounds on ARN KISS neuron firing in acute brain slices from diestrous female mice. (A) Robust excitatory effect of 90 seconds puff of 40 µM serotonin on firing rate of a middle ARN kisspeptin neuron. (B) Inhibitory effect of 90 seconds puff of 60 µM serotonin on firing rate of a caudal ARN kisspeptin neuron initially stimulated to fire by a 200 nM puff of NKB. (C) Rostral ARN kisspeptin neuron not responding to 60 µM puffs of serotonin but later activated by NKB. (D) Summary of percentage of rostral (rARN), middle (mARN), and caudal (cARN) kisspeptin neurons excited or inhibited by serotonin. (E) Caudal ARN kisspeptin neuron activated by serotonin (40 µM) in the absence and presence of methiothepin (100 µM). (F) Middle ARN kisspeptin neuron in which the excitatory effect of serotonin (40 µM) is blocked by methiothepin (100 µM). (G) Middle ARN kisspeptin neuron activated by serotonin (40 µM) in the absence and presence of SB228357 (100 µM). (H) Caudal ARN kisspeptin neuron in which zacopride (2 µM) facilitates serotonin excitation. (I) Whole-cell recording from an ARN KISS neuron in the continuous presence of TTX, CNQX, DAP5, and bicuculline, showing depolarization during bath application of 5-HT (30 µM). (J) Summary of mean membrane potential measured predrug, during 5-HT application, and following wash ( P < .001; n = 4 animals).

Article Snippet: Stock solutions of GABAzine (SR95531, 5 mM, Tocris, UK), CNQX (10 mM, Tocris, UK), and DAP5, (50 mM, Tocris, UK) were prepared with Milli-Q water or NaOH for DAP5.

Techniques: Membrane