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In the present study, the SAN trip containing "the central nodal region" is located by tagging super vena cava with black fine tread as a mark in right atrium of rabbit. The beating pacemaker cell of the SAN is spontaneously isolated by two dIfferent enzymatic digestions. Hyperpolarization-activated pacemaker current is recorded by using whole-cell perforated patch technique in SAN cells. The results show that SAN tissue which include the "central nodal region" is shaped with superior wide and inferior narrow, and with an volume size that is approximately 6.25mm×2.23mm×0.26mm or 6mm×2mm×0.1mm—6mm×2mm×3.3mm. SAN cells are long thin spindle-shaped with faint cross-striations. They are measured with (65.3±7.8)μm in length and (7.2±3.1)μm in width. In normal Tyrode's solution, the typical SAN cells which are 2%—5% in yielded all of them are recovered spontaneous beating with frequency of (187.3±5.6) times/min. According to voltage clamp protocol, If is elicited as the inward ramp current, and becomes larger as soon as is activated more rapidly with the increasingly negative potential. The tail current of If has similar characteristics in same conditions. The current-voltage curve of If and tail is appearing as a typical horizontal "Y" according to their current density and clamp potential. It could be identIfied that the isolated SAN cells have typical morphological properties and activity, therefore could be completely elicited inward ramp If, the fact could contribute to the dIfferent investigations on If in SAN cells in cardiovascular areas., authors=WANG Teng1, WU Pingya2, FAN Xinrong1, CHEN Yongjun1, CHEN Hui1, HUANG Congxin1, authorsList=WANG Teng;WU Pingya;FAN Xinrong;CHEN Yongjun;CHEN Hui;HUANG Congxin, authorCompany=1. Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan 430060, China;2. 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Current,If)的特征.通过打结定位法确定了带有"中心结区"的家兔SAN组织条,双酶法分离SAN细胞,利用全细胞穿孔膜片钳技术记录SAN细胞的If.结果显示,SAN组织为上宽下窄的枣核型,区域体积约为6.25mm×2.23mm×0.26mm,或6mm×2mm×0.1mm—6mm×2mm×3.3mm.SAN细胞成狭长梭形,无清晰横纹,细胞长(65.3±7.8)μm,宽为(7.2±3.1)μm.在正常Tyrode液中,可恢复规则自律性搏动,频率为(187.3±5.6)次/min,典型SAN细胞数量占整个分离细胞的2%—5%.根据电压钳制方案,If电流被引出,呈内向性斜坡状,随着钳制电压降低,起搏电流逐渐增大,并随钳制时间延长,倾斜度也增大.If带有明显尾电流,也随着超级化激活电位降低,逐渐增大.根据二者电流大小拟合的电流-电压曲线,呈典型横向Y形.本法获得SAN细胞形态典型,状态和活性良好,能够记录具有特征性的If,可供心血管领域有关针对SAN细胞If研究的技术借鉴., authors=王腾1, 吴平亚2, 范新荣1, 陈勇军1, 陈卉1, 黄从新1, authorsList=王腾;吴平亚;范新荣;陈勇军;陈卉;黄从新, authorCompany=1. 武汉大学人民医院心内科,武汉 430060;2. 武汉市洪山区张家湾社区卫生服务中心,武汉 430070, correspAuthors=null, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=QlvL33gmf6nwWnYpWYCyNg==, pdfFileSize=2235987, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, 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兔窦房结细胞的分离方法及其起搏电流特征性探讨
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王腾;吴平亚;范新荣;陈勇军;陈卉;黄从新
科技导报 | 研究论文 2011,29(23): 29-32
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科技导报 |研究论文 2011 , 29 (23) : 29 -32
兔窦房结细胞的分离方法及其起搏电流特征性探讨
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王腾;吴平亚;范新荣;陈勇军;陈卉;黄从新
作者信息
Sino-atrial Node Cell Dissociation Method and Pacemaker Current Characteristics in Rabbits
WANG Teng;WU Pingya;FAN Xinrong;CHEN Yongjun;CHEN Hui;HUANG Congxin
Affiliations
    1. Department of Cardiology, Renmin Hospital of Wuhan University, Wuhan 430060, China;2. Community Health Service Center, Zhangjiangwan Street, Hongshan District, Wuhan 430070, China
出版时间: 2011-08-18 doi: 10.3981/j.issn.1000-7857.2011.23.002
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探讨了家兔窦房结(Sino-Atrial Node,SAN)细胞的分离方法及其形态和起搏电流(Funny Current,If)的特征.通过打结定位法确定了带有"中心结区"的家兔SAN组织条,双酶法分离SAN细胞,利用全细胞穿孔膜片钳技术记录SAN细胞的If.结果显示,SAN组织为上宽下窄的枣核型,区域体积约为6.25mm×2.23mm×0.26mm,或6mm×2mm×0.1mm—6mm×2mm×3.3mm.SAN细胞成狭长梭形,无清晰横纹,细胞长(65.3±7.8)μm,宽为(7.2±3.1)μm.在正常Tyrode液中,可恢复规则自律性搏动,频率为(187.3±5.6)次/min,典型SAN细胞数量占整个分离细胞的2%—5%.根据电压钳制方案,If电流被引出,呈内向性斜坡状,随着钳制电压降低,起搏电流逐渐增大,并随钳制时间延长,倾斜度也增大.If带有明显尾电流,也随着超级化激活电位降低,逐渐增大.根据二者电流大小拟合的电流-电压曲线,呈典型横向Y形.本法获得SAN细胞形态典型,状态和活性良好,能够记录具有特征性的If,可供心血管领域有关针对SAN细胞If研究的技术借鉴.
窦房结  /  细胞形态学  /  超级化激活起搏电流
The isolation means and properties of morphology and pacemaker current If in Sino-Atrial Node (SAN) cells of rabbits are investigated. In the present study, the SAN trip containing "the central nodal region" is located by tagging super vena cava with black fine tread as a mark in right atrium of rabbit. The beating pacemaker cell of the SAN is spontaneously isolated by two dIfferent enzymatic digestions. Hyperpolarization-activated pacemaker current is recorded by using whole-cell perforated patch technique in SAN cells. The results show that SAN tissue which include the "central nodal region" is shaped with superior wide and inferior narrow, and with an volume size that is approximately 6.25mm×2.23mm×0.26mm or 6mm×2mm×0.1mm—6mm×2mm×3.3mm. SAN cells are long thin spindle-shaped with faint cross-striations. They are measured with (65.3±7.8)μm in length and (7.2±3.1)μm in width. In normal Tyrode's solution, the typical SAN cells which are 2%—5% in yielded all of them are recovered spontaneous beating with frequency of (187.3±5.6) times/min. According to voltage clamp protocol, If is elicited as the inward ramp current, and becomes larger as soon as is activated more rapidly with the increasingly negative potential. The tail current of If has similar characteristics in same conditions. The current-voltage curve of If and tail is appearing as a typical horizontal "Y" according to their current density and clamp potential. It could be identIfied that the isolated SAN cells have typical morphological properties and activity, therefore could be completely elicited inward ramp If, the fact could contribute to the dIfferent investigations on If in SAN cells in cardiovascular areas.
sino-atrial node  /  cell morphology  /  hyperpolarization-activated pacemaker current
王腾;吴平亚;范新荣;陈勇军;陈卉;黄从新. 兔窦房结细胞的分离方法及其起搏电流特征性探讨. 科技导报, 2011 , 29 (23) : 29 -32 . DOI: 10.3981/j.issn.1000-7857.2011.23.002
WANG Teng;WU Pingya;FAN Xinrong;CHEN Yongjun;CHEN Hui;HUANG Congxin. Sino-atrial Node Cell Dissociation Method and Pacemaker Current Characteristics in Rabbits[J]. Science & Technology Review, 2011 , 29 (23) : 29 -32 . DOI: 10.3981/j.issn.1000-7857.2011.23.002

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doi: 10.3981/j.issn.1000-7857.2011.23.002
  • 接收时间:2011-06-08
  • 首发时间:2011-08-18
  • 出版时间:2011-08-18
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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
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