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Acute hippocampal slice preparation and electrophysiological analysis

TL Tian Lan
YL Ye Li
CF Cuiqin Fan
LW Liyan Wang
WW Wenjing Wang
SC Shihong Chen
SY Shu Yan Yu
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Rats were anesthetized using 2.5% isoflurane and decapitated. The brain was extracted, blocked, placed on a vibrating slicer, and immersed in a cold solution containing (in mM) 119 choline chloride, 30 Glucose, 26 NaHCO3, 7 MgSO4, 2.5 KCl, 1 NaH2PO4, 1 CaCl2, 3 sodiumpyruvate, 1.3 sodium L-ascorbate, 1 kynurenicacid, and saturated with 95% O2/5% CO2. Slices were then transferred as quickly as possible to a recovery solution containing (in mM) 85 NaCl, 24 NaHCO3, 4 MgCl2, 2.5 KCl, 1.25 NaH2PO4, 0.5 CaCl2, 25 glucose and 50 sucrose and allowed to recover for 30 min at 30 °C. The glass micropipettes (4–6 MΩ) were filled with an internal solution containing (in mM) 130 CsMeSO4, 10 CsCl, 4 NaCl, 1 MgCl2, 5 MgATP, 5 EGTA, 10 HEPES, 0.5 Na3GTP, 10 phosphocreatine and 4 QX-314, with a pH of 7.35. During whole-cell clamp patch recordings, slices were continuously perfused with an artificial cerebral spinal fluid (ACSF) contained (in mM) 120 NaCl, 3.5 KCl, 2.5 CaCl2, 1.3 MgSO4, 1.25 NaH2PO4, 26 NaHCO3 and 10 glucose.

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