培养的小鼠小肠和胃Cajal间质细胞起搏电流的特性
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朴莲花, 张 扬, 黄 旭, 李春晖, 高 玲, 许文燮
Cajal间质细胞;起搏电流;钙调蛋白;胞内钙,朴莲花,张扬,黄旭,高玲,许文燮,朴莲花,李春晖,朴莲花,通讯作者:,Characteristicsofthepacemakercurrentinculturedinterstitialc
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朴莲花, 张扬, 黄旭, 高玲, 许文燮, 上海交通大学医学院生理学教研室 上海市 200240
朴莲花, 李春晖, 延边大学医学院生理学教研室 吉林省延吉市 133000
朴莲花, 2003级生理学专业硕士生, 主要从事消化道平滑肌动力学研究.
国家自然科学基金资助项目, No. 30160028, No. 30360031
通讯作者: 许文燮, 200240, 上海市东川路800号, 上海交通大学文选医学楼328室. wenxiexu@sjtu.edu.cn
电话: 021-34205639
收稿日期: 2006-08-24 接受日期: 2006-09-20
Characteristics of the pacemaker current in cultured interstitial cells of Cajal from murine stomach and small intestine
Lian-Hua Piao, Yang Zhang, Xu Huang, Chun-Hui Li, Ling Gao, Wen-Xie Xu
Lian-Hua Piao, Yang Zhang, Xu Huang, Ling Gao, Wen-Xie Xu,Department of Physiology, Medical College of Shanghai Jiao Tong University, Shanghai 200240, China
Lian-Hua Piao, Chun-Hui Li,Department of Physiology, Medical College, Yanbian University, Yanji 133000, Jinlin Province, China
Supported byNational Natural Science Foundation of China, No. 30160028, and No. 30360031
Correspondence to:Professor Wen-Xie Xu, Room 328, Wenxuan Medical Building, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China. wenxiexu@sjtu.edu.cn
Received:2006-08-24 Accepted: 2006-09-20
Abstract
AIM: To investigate the characteristics of the pacemaker current in cultured interstitial cells of Cajal (ICC) from murine stomach and small intestine.
METHODS: ICC was isolated from murine stomach and small intestine and then cultured in the medium with stem cell factors. After 72 hours, ICC was identified by immunohistochemical technique with monoclonal antibody for c-kit protein labelling and the pacemaker currents in gastric and intestinal ICC were recorded under the conventional whole-cell patch-clamp configuration. When the concentration of calcium-chelating agent EGTA was increased from 0.1 to 10 mmol/L, the effects of low intracellular calcium on the pacemaker currents were observed.
RESULTS: At the 72nd hour, 2 to 4 projections were stacked out from the body of ICC, which interconnected with neighboring cells and formed cell networks under light microscope. Immunohistochemistry showed that c-kit protein was positively expressed on the surface of ICC. Under the conventional whole-cell configuration, the spontaneous and rhythmic inward currents (pacemaker currents) were elicited in cultured gastric and small intestinal ICC, respectively, as the membrane potential was held at -60 mV. There were differences in the amplitudes and frequencies between single ICC and ICC networks or between gastric ICC and small intestinal ICC. Under the conventional whole-cell configuration, an inward current was activated in ICC by increasing the concentration of EGTA in pipette solution W-7, a calmodulin-inhibitor, which activated the inward current and increased the amplitude of pacemaker currents in ICC.
CONCLUSION: The frequencies of the pacemaker currents from gastric and small intestinal ICC are very similar to the frequencies of rhythmic contraction in murine gastric and small intestinal smooth muscles; ICC network generates higher- amplitude, stabler and more rhythmic pacemaker currents than single ICC does; low concentration of intracellular free calcium is an important factor for generating pacemaker current; calmodulin is involved in the inhibitory regulation of the pacemaker current ......
朴莲花, 张扬, 黄旭, 高玲, 许文燮, 上海交通大学医学院生理学教研室 上海市 200240
朴莲花, 李春晖, 延边大学医学院生理学教研室 吉林省延吉市 133000
朴莲花, 2003级生理学专业硕士生, 主要从事消化道平滑肌动力学研究.
国家自然科学基金资助项目, No. 30160028, No. 30360031
通讯作者: 许文燮, 200240, 上海市东川路800号, 上海交通大学文选医学楼328室. wenxiexu@sjtu.edu.cn
电话: 021-34205639
收稿日期: 2006-08-24 接受日期: 2006-09-20
Characteristics of the pacemaker current in cultured interstitial cells of Cajal from murine stomach and small intestine
Lian-Hua Piao, Yang Zhang, Xu Huang, Chun-Hui Li, Ling Gao, Wen-Xie Xu
Lian-Hua Piao, Yang Zhang, Xu Huang, Ling Gao, Wen-Xie Xu,Department of Physiology, Medical College of Shanghai Jiao Tong University, Shanghai 200240, China
Lian-Hua Piao, Chun-Hui Li,Department of Physiology, Medical College, Yanbian University, Yanji 133000, Jinlin Province, China
Supported byNational Natural Science Foundation of China, No. 30160028, and No. 30360031
Correspondence to:Professor Wen-Xie Xu, Room 328, Wenxuan Medical Building, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China. wenxiexu@sjtu.edu.cn
Received:2006-08-24 Accepted: 2006-09-20
Abstract
AIM: To investigate the characteristics of the pacemaker current in cultured interstitial cells of Cajal (ICC) from murine stomach and small intestine.
METHODS: ICC was isolated from murine stomach and small intestine and then cultured in the medium with stem cell factors. After 72 hours, ICC was identified by immunohistochemical technique with monoclonal antibody for c-kit protein labelling and the pacemaker currents in gastric and intestinal ICC were recorded under the conventional whole-cell patch-clamp configuration. When the concentration of calcium-chelating agent EGTA was increased from 0.1 to 10 mmol/L, the effects of low intracellular calcium on the pacemaker currents were observed.
RESULTS: At the 72nd hour, 2 to 4 projections were stacked out from the body of ICC, which interconnected with neighboring cells and formed cell networks under light microscope. Immunohistochemistry showed that c-kit protein was positively expressed on the surface of ICC. Under the conventional whole-cell configuration, the spontaneous and rhythmic inward currents (pacemaker currents) were elicited in cultured gastric and small intestinal ICC, respectively, as the membrane potential was held at -60 mV. There were differences in the amplitudes and frequencies between single ICC and ICC networks or between gastric ICC and small intestinal ICC. Under the conventional whole-cell configuration, an inward current was activated in ICC by increasing the concentration of EGTA in pipette solution W-7, a calmodulin-inhibitor, which activated the inward current and increased the amplitude of pacemaker currents in ICC.
CONCLUSION: The frequencies of the pacemaker currents from gastric and small intestinal ICC are very similar to the frequencies of rhythmic contraction in murine gastric and small intestinal smooth muscles; ICC network generates higher- amplitude, stabler and more rhythmic pacemaker currents than single ICC does; low concentration of intracellular free calcium is an important factor for generating pacemaker current; calmodulin is involved in the inhibitory regulation of the pacemaker current ......
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