Journal: The Journal of Clinical Investigation

Article Title: Regularizing firing patterns of rat subthalamic neurons ameliorates parkinsonian motor deficits

doi: 10.1172/JCI99986

Figure Lengend Snippet: (A) Microscope image of a STN neuron (indicated by arrowheads) recorded in a brain slice and labeled with biocytin after patch-clamp recording. (B) Histamine (10 μM) shifted the conductance-voltage curve of recorded STN neurons (at –90 mV and –100 mV, n = 5). The conductance curve was converted from the whole-cell currents recorded from –50 to –120 mV and was fitted by Boltzmann function. Note that the conductance exhibited a significant feature of hyperpolarization activation and histamine reduced the voltage required for half-maximal activation (V1/2, n = 5). (C) Histamine increased the inward rectification (sag) in response to an 80 pA hyperpolarizing current pulse. ZD7288 (50 μM), a selective blocker for the HCN channel, abolished the depolarizing sag in both the absence and presence of histamine (n = 8). (D) Histamine elicited an inward current in a STN neuron, and ranitidine (1 μM), a selective antagonist for the H2 receptor, or ZD7288 (50 μM) totally blocked the current induced by histamine (n = 8). (E and F) PSTHs, scatter plots of ISI series, autocorrelation histograms, and an ISI histogram of discharges of a recorded STN neuron show the histamine-induced changes in firing rate and firing pattern in the absence and presence of ranitidine and ZD7288 (1 μM, respectively). (G) Group data show that histamine significantly decreased the CV of ISIs, whereas ZD7288 remarkably increased the CV and blocked the histamine-induced decrease in CV (n = 30). Data are represented as mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001, 2-tailed paired t test (B) or 1-way ANOVA with Newman-Keuls post hoc test (C, D, and G).

Article Snippet: Histamine (1 μg; MilliporeSigma), high K + (0.75 μg KCl), 2-PyEA (a selective agonist for the H1 receptor; 1 μg; MilliporeSigma), dimaprit (a selective agonist for the H2 receptor; 2 μg; Tocris), R-(-)-α-methylhistamine (a selective agonist for the H3 receptor; 1.5 μg; Tocris), VUF8430 (a selective agonist for the H4 receptor; 3 μg; Tocris), mepyramine (a selective antagonist for the H1 receptor; 4 μg; Tocris), ranitidine (a selective antagonist for the H2 receptor; 3.5 μg; Tocris), JNJ5207852 (a selective antagonist for the H3 receptor; 2 μg; Tocris), JNJ7777120 (a selective agonist for the H4 receptor; 2.5 μg; Tocris), ZD7288 (a selective blocker for the HCN channel; 3 μg; Tocris), or normal saline (0.9% NaCl) was unilaterally or bilaterally microinjected with a Hamilton syringe or syringes (0.5 μl each side, lasting 2 minutes).

Techniques: Microscopy, Slice Preparation, Labeling, Patch Clamp, Activation Assay

Journal: The Journal of Clinical Investigation

Article Title: Regularizing firing patterns of rat subthalamic neurons ameliorates parkinsonian motor deficits

doi: 10.1172/JCI99986

Figure Lengend Snippet: Effects of microinjection of histamine (1 μg), dimaprit (a selective agonist for H2 receptor, 2 μg), ranitidine (a selective antagonist for H2 receptor, 3.5 μg), and ZD7288 (a selective blocker for HCN channel, 3 μg) into the STN on turning behavior (A, n = 12), adhesive-removal test (B, n = 10), and locomotor footprints (C, n = 10) in PD rats. Dimaprit mimicked the histamine-induced decrease in turnings, shortened time for removing an adhesive strip from forelimb contralateral to the lesion, and enlarged bilateral stride lengths, whereas ranitidine and ZD7288 significantly increased the turnings, prolonged contralesional adhesive-removal time, and shortened bilateral stride lengths. ZD7288 also abolished the histamine-induced amelioration of turning, adhesive removal, and locomotor behaviors in PD rats. Data are represented as median (horizontal bar) with 25th–75th (box) and 5th–95th (whiskers) percentiles or mean ± SEM. **P < 0.01; ***P < 0.001, 1-way (A and C, stride width) or 2-way ANOVA (B and C, stride length) with Newman-Keuls post hoc test.

Article Snippet: Histamine (1 μg; MilliporeSigma), high K + (0.75 μg KCl), 2-PyEA (a selective agonist for the H1 receptor; 1 μg; MilliporeSigma), dimaprit (a selective agonist for the H2 receptor; 2 μg; Tocris), R-(-)-α-methylhistamine (a selective agonist for the H3 receptor; 1.5 μg; Tocris), VUF8430 (a selective agonist for the H4 receptor; 3 μg; Tocris), mepyramine (a selective antagonist for the H1 receptor; 4 μg; Tocris), ranitidine (a selective antagonist for the H2 receptor; 3.5 μg; Tocris), JNJ5207852 (a selective antagonist for the H3 receptor; 2 μg; Tocris), JNJ7777120 (a selective agonist for the H4 receptor; 2.5 μg; Tocris), ZD7288 (a selective blocker for the HCN channel; 3 μg; Tocris), or normal saline (0.9% NaCl) was unilaterally or bilaterally microinjected with a Hamilton syringe or syringes (0.5 μl each side, lasting 2 minutes).

Techniques: Microinjection, Adhesive, Stripping Membranes

Journal: Scientific reports

Article Title: ZD7288 ameliorates long-term cerebral ischemia reperfusion-induced deterioration of spatial memory by regulating the expressions of NMDA receptor subunits in rats.

doi: 10.1038/s41598-025-03172-8

Figure Lengend Snippet: Fig. 2. The neuroprotective effects of ZD7288 on IR-induced injury. ZD7288 or MK-801 was administrated by lateral ventricular injection for 2 h before MCAO. (A) The Zea Longa scores were performed after the rats recovered from the 2-hour ischemic period. (B) The mNSS was evaluated following the 2-hour ischemia recovery. (C) A representative image of the TTC staining. (D) The quantification data from the TTC staining. *P < 0.05, n = 10.

Article Snippet: ZD7288 (SC-361419 A, Santa Cruz Biotechnology, Dallas, USA); MK-801 (Med Chem Express China, Shanghai, China); Isoflurane (RWD Life Science, Shenzhen, China); Nylon fishing line, diameter 0.23 mm (Rikimaru, Mihama, Japan); Injection catheter (RWD Life Science, Shenzhen, China); 2,3,5-triphenyl tetrazolium chloride (TTC, Sigma-Aldrich, St. Louis, USA); PrimeScriptTM RT reagent Kit (TaKaRa Bio, Dalian, China); SYBR® premix Ex TaqTM II (TaKaRa Bio, Dalian, China).

Techniques: Injection, Staining

Journal: Scientific reports

Article Title: ZD7288 ameliorates long-term cerebral ischemia reperfusion-induced deterioration of spatial memory by regulating the expressions of NMDA receptor subunits in rats.

doi: 10.1038/s41598-025-03172-8

Figure Lengend Snippet: Fig. 3. ZD7288 ameliorates the spatial memory after IR. Rats were administered ZD7288 or MK-801 2 h before MCAO. The place navigation period spanned from Day 1 to Day 5, with the platform removed on Day 6. (A) Representative images of swimming paths. (B) Average swimming speed from Day 1 to Day 5. (C) Escape latency from Day 1 to Day 5. (D) Time spent in the target quadrant on Day 6. (E) Number of platform crossings on Day 6. *P < 0.05, n = 10.

Article Snippet: ZD7288 (SC-361419 A, Santa Cruz Biotechnology, Dallas, USA); MK-801 (Med Chem Express China, Shanghai, China); Isoflurane (RWD Life Science, Shenzhen, China); Nylon fishing line, diameter 0.23 mm (Rikimaru, Mihama, Japan); Injection catheter (RWD Life Science, Shenzhen, China); 2,3,5-triphenyl tetrazolium chloride (TTC, Sigma-Aldrich, St. Louis, USA); PrimeScriptTM RT reagent Kit (TaKaRa Bio, Dalian, China); SYBR® premix Ex TaqTM II (TaKaRa Bio, Dalian, China).

Techniques:

Journal: Scientific reports

Article Title: ZD7288 ameliorates long-term cerebral ischemia reperfusion-induced deterioration of spatial memory by regulating the expressions of NMDA receptor subunits in rats.

doi: 10.1038/s41598-025-03172-8

Figure Lengend Snippet: Fig. 4. ZD7288 improves the morphology of neurons in different regions of the hippocampus after IR. ZD7288 or MK-801 was administered via lateral ventricular injection for 2 h before MCAO. (A–C) The representative images of HE staining in various regions of the hippocampus, captured at 200× and 400× magnification. Black arrows indicate normal neurons, while red arrows denote ischemic neurons. (D–F): Cells were counted in the CA1, CA3, and DG regions at 200× magnification, which were used to calculate the percentage of damaged neurons. *P < 0.05, n = 3. ns, not significant.

Article Snippet: ZD7288 (SC-361419 A, Santa Cruz Biotechnology, Dallas, USA); MK-801 (Med Chem Express China, Shanghai, China); Isoflurane (RWD Life Science, Shenzhen, China); Nylon fishing line, diameter 0.23 mm (Rikimaru, Mihama, Japan); Injection catheter (RWD Life Science, Shenzhen, China); 2,3,5-triphenyl tetrazolium chloride (TTC, Sigma-Aldrich, St. Louis, USA); PrimeScriptTM RT reagent Kit (TaKaRa Bio, Dalian, China); SYBR® premix Ex TaqTM II (TaKaRa Bio, Dalian, China).

Techniques: Injection, Staining

Journal: Scientific reports

Article Title: ZD7288 ameliorates long-term cerebral ischemia reperfusion-induced deterioration of spatial memory by regulating the expressions of NMDA receptor subunits in rats.

doi: 10.1038/s41598-025-03172-8

Figure Lengend Snippet: Fig. 5. ZD7288 reduces the elevation of HCN1 and HCN2 induced by IR. After completing the MWM, the expressions of HCN1 and HCN2 in the ischemic hippocampus were detected. (A) The representative pictures of immunoblotting. (B, C) The quantification data for HCN1. (D, E) The quantification data for HCN2. Original images of HCN1 and HCN2 are available at Supplemental Material. *P < 0.05, n = 4.

Article Snippet: ZD7288 (SC-361419 A, Santa Cruz Biotechnology, Dallas, USA); MK-801 (Med Chem Express China, Shanghai, China); Isoflurane (RWD Life Science, Shenzhen, China); Nylon fishing line, diameter 0.23 mm (Rikimaru, Mihama, Japan); Injection catheter (RWD Life Science, Shenzhen, China); 2,3,5-triphenyl tetrazolium chloride (TTC, Sigma-Aldrich, St. Louis, USA); PrimeScriptTM RT reagent Kit (TaKaRa Bio, Dalian, China); SYBR® premix Ex TaqTM II (TaKaRa Bio, Dalian, China).

Techniques: Western Blot

Journal: Scientific reports

Article Title: ZD7288 ameliorates long-term cerebral ischemia reperfusion-induced deterioration of spatial memory by regulating the expressions of NMDA receptor subunits in rats.

doi: 10.1038/s41598-025-03172-8

Figure Lengend Snippet: Fig. 6. ZD7288 regulates the total expression of NMDAR subunits in the ischemic hippocampus after IR. ZD7288 or MK-801 was administered via lateral ventricular injection for 2 h before MCAO. (A) The representative immunoblotting pictures of the NMDAR subunits in the ischemic hippocampus. (B) Quantitative data for protein and mRNA levels of NR1 and p-NR1. (C) Quantitative data for protein and mRNA levels of NR2A and p-NR2A. (D) Quantitative data for protein and mRNA levels of NR2B and p-NR2B. (E) Quantitative data for protein and mRNA levels of NR3A. (F) Quantitative data for protein and mRNA levels of NR3B. Original images of NMDAR subunits are available at Supplemental Material. *P < 0.05, n = 4.

Article Snippet: ZD7288 (SC-361419 A, Santa Cruz Biotechnology, Dallas, USA); MK-801 (Med Chem Express China, Shanghai, China); Isoflurane (RWD Life Science, Shenzhen, China); Nylon fishing line, diameter 0.23 mm (Rikimaru, Mihama, Japan); Injection catheter (RWD Life Science, Shenzhen, China); 2,3,5-triphenyl tetrazolium chloride (TTC, Sigma-Aldrich, St. Louis, USA); PrimeScriptTM RT reagent Kit (TaKaRa Bio, Dalian, China); SYBR® premix Ex TaqTM II (TaKaRa Bio, Dalian, China).

Techniques: Expressing, Injection, Western Blot

Journal: Scientific reports

Article Title: ZD7288 ameliorates long-term cerebral ischemia reperfusion-induced deterioration of spatial memory by regulating the expressions of NMDA receptor subunits in rats.

doi: 10.1038/s41598-025-03172-8

Figure Lengend Snippet: Fig. 7. ZD7288 regulates the expressions of NMDAR subunits in the different hippocampus regions after IR. After completing the MWM, the expressions of NMDAR subunits in the CA1, CA3, and DG regions were tested. (A–H): Immunohistochemical staining and quantification data for the expression of NR1, NR2A, NR2B, NR3A, p-NR1, p-NR2A, p-NR2B, and NR3B in the CA1, CA3, and DG regions. *P < 0.05, n = 3.

Article Snippet: ZD7288 (SC-361419 A, Santa Cruz Biotechnology, Dallas, USA); MK-801 (Med Chem Express China, Shanghai, China); Isoflurane (RWD Life Science, Shenzhen, China); Nylon fishing line, diameter 0.23 mm (Rikimaru, Mihama, Japan); Injection catheter (RWD Life Science, Shenzhen, China); 2,3,5-triphenyl tetrazolium chloride (TTC, Sigma-Aldrich, St. Louis, USA); PrimeScriptTM RT reagent Kit (TaKaRa Bio, Dalian, China); SYBR® premix Ex TaqTM II (TaKaRa Bio, Dalian, China).

Techniques: Immunohistochemical staining, Staining, Expressing