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3 protocols using kb r7943

1

Inhibition of NCX Reverse Mode Activity

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We used SN-6 (2-[[4-[(4-Nitrophenyl)methoxy]phenyl]methyl]-4-thiazolidine carboxylic acid ethyl ester) and KB-R7943 (2-[2-[4-(4-Nitrobenzyloxy)phenyl]ethyl]isothiourea mesylate) potent inhibitors of the NCX reverse mode activity for NCX1 and NCX3, respectively, which were purchased from R&D System (Minneapolis, MN, USA). SN-6 and KB-R7943 were dissolved in dimethyl sulfonic acid (0.1%) and phosphate-buffered saline (pH 7.4) using sonication (80 kHz, 100% power). The solutions containing either SN-6 (0, 3, 10 mg/kg, p.o.) or KB-R7943 (0, 3, 10 mg/kg, p.o.) were freshly prepared before their administration, as previously described [29 (link),30 (link)]. After four weeks of training, animals were randomly assigned into eight groups (n= 12) including female KB-R7943-3 (KB-R7943, 3 mg/kg), female KB-R7943-10 (KB-R7943, 10 mg/kg), male KB-R7943-3 (KB-R7943, 3 mg/kg), male KB-R7943-10 (KB-R7943, 10 mg/kg), female SN-6-3 (KB-R7943, 3 mg/kg), female SN-6-10 (SN-6, 10 mg/kg), male SN-6-3 (SN-6, 3 mg/kg), and male SN-6-10 (SN-6, 10 mg/kg). On Mondays, animals received the vehicle; on Wednesdays, animals received the tested drug, either SN-6 or KB-R7943; and on Fridays, they were examined to see the long-lasting effects of the drugs.
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2

Probing NCX Role in AWS Pathophysiology

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To probe the role of NCX in the AWS pathophysiology, we used SN-6 (2-[[4-[(4-nitrophenyl)methoxy]phenyl]methyl]-4-thiazoli dinecarboxylic acid ethyl ester, R&D Systems, Minneapolis, MN, USA) and KB-R7943 (2-[2-[4-(4-nitrobenzyloxy)phenyl]ethyl] isothioureamethanesulfonate, R&D Systems). SN-6 preferentially blocks NCX1rev activity, while KB-7943 inhibits NCX3rev more potently than its forward mode [40 (link)–42 (link)]. SN-6 and KB-R7943 were dissolved in dimethyl sulfonic acid (0.1%) and phosphate-buffered saline (pH 7.4) using sonication (80 kHz, 100% power). The solutions containing SN-6 or KB-R7943 were filtered before focal microinjections within the IC at 5 μg/hemisphere; this dose was chosen based on our published reports [24 (link),43 (link)].
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3

Pharmacological Agents in Neuronal Signaling

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The following chemicals were products of R&D Systems: AVP, tetrodotoxin (TTX), kynurenic acid, picrotoxin, SR49059, TASP0390325, tolvaptan, GDP-β-S, U73122, U73343, heparin, thapsigargin, chelerythrine, bisindolylmaleimide II (Bis II), KB-R7943, capsazepine, AMG9810, AMG1629, capsaicin, ML 133, ML 297 and SCH23390. Dioctanoyl phosphatidylinositol 4,5-bisphosphate (dic8-PIP2) was purchased from Echelon Biosciences. Drugs were initially prepared in stock solution, aliquoted and stored at −20°C. For those chemicals requiring dimethyl sulfoxide (DMSO) as a solvent, the concentration of DMSO was less than 0.1%. This concentration of DMSO either in the recording pipettes or in the bath had no significant effects on neuronal activity.
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