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目的介绍一种在微动脉段标本上进行全细胞膜片钳记录的技术。方法在离体豚鼠耳蜗螺旋动脉(SMA)、小脑前下动脉(AICA)和肠系膜动脉(MA)分支微动脉(直径小于100μm)的原位平滑肌细胞上,应用全细胞膜片钳技术。结果微动脉段上平滑肌细胞静息膜电位平均值在-25～-37 mV之间,细胞膜电容(Cinput)平均值在70～250 pF之间,均高于消化分离的单个平滑肌细胞。应用缝隙连接阻断剂DPBA(100μmol/L)后,SMA、AICA和MA微动脉段上平滑肌细胞膜电阻(Rinput)分别为(4 937±741)MΩ(n=12)、(3 703±367)MΩ(n=8)和(3336±479)MΩ(n=12),细胞Cinput分别为(4.5±0.2)pF(n=9)、(7.1±0.7)pF(n=5)和(9.6±0.9)pF(n=7),与单个平滑肌细胞相似,表明微动脉细胞间存在着丰富的缝隙连接。结论微动脉段标本全细胞膜片钳记录技术适用于微动脉细胞间缝隙连接、神经递质和药物对微动脉作用机制的研究。
Objective To introduce a technique of whole-cell patch-clamp recording on micro-arterial specimens. Methods Whole-cell patch-clamp technique was applied to in situ smooth muscle cells of isolated guinea pig cochlear spiral arteries (SMA), anterior communicating artery (AICA) and mesenteric artery (MA) arterioles (diameter less than 100 μm) Results The average resting membrane potential of smooth muscle cells in the arterioles ranged from -25 to -37 mV and the average Cinput values ranged from 70 to 250 pF, which were higher than that of single smooth muscle cells digested and separated. The membrane resistance (Rinput) of smooth muscle cells in SMA, AICA and MA arterioles were (4 937 ± 741) MΩ (n = 12) and (3 703 ± 367) μmol / L after DPBA (100 μmol / Cinput were (4.5 ± 0.2) pF (n = 9), (7.1 ± 0.7) pF (n = 5) and (9.6 ± 0.9) pF (n = 7), similar to single smooth muscle cells, suggesting abundant gap junctions between arteriolar cells. Conclusion The whole-cell patch-clamp recording technique of micro-arterial section is suitable for the study of the mechanism of micro-arteries connection between arteriole gap junction, neurotransmitter and drug.