谷瑞民
应用单通道膜片钳技术研究肾脏离子通道活性的调控机制。
个性化签名
- 姓名:谷瑞民
- 目前身份:
- 担任导师情况:
- 学位:
-
学术头衔:
博士生导师
- 职称:-
-
学科领域:
生理学
- 研究兴趣:应用单通道膜片钳技术研究肾脏离子通道活性的调控机制。
谷瑞民教授,博士生导师,中国生理科学会理事,中国生理科学会循环肾脏专业委员会委员。1975年毕业留校任教于基础医学院生理学教研室,历任助教,讲师,副教授,教授,教研室副主任,主任,黑龙江重点学科(生理学)学科带头人,谷瑞民教授作为访问学者于1991年10月-1992年10月、1997年3月-1998年3月由国家教委会派出,分别在德国基尔大学和雷根斯堡大学生理研究所研修。1991年1月至2002年3月,在美国纽约医学院做研究工作。
谷瑞民教授在90年代前,主要从事痛与锐痛机制的研究,应用细胞外记录方法,观察不同化学物质及神经递质对大鼠中枢神经核团痛兴奋神经和痛抑制神经元的作用,探讨痛相关神经元的调控机制。研究项目“几类中枢递质在镇痛中的作用及其机理分析”获黑龙江省科学技术进步三等奖及黑龙江省教育委员会科学技术进步一等奖。合作项目“带肌蒂大转子筋膜再生软骨的实验研究获黑龙江省科学技术进步三等奖,带臀中肌蒂大转子移植重建股骨头治疗重症股骨头缺血性坏死技术获国家发明三等奖。
在近10年国内外研究工作期间,应用单通道膜片钳技术研究肾脏离子通道活性的调控机制。主要研究范围包括1、花生四烯酸及代谢产物20-HETE对髓袢升支粗段管周膜钾通道的影响及作用的研究。2、酪氨酸蛋白激酶和磷酸化酶对ROMK1的作用研究 3、酪氨酸蛋白激酶和磷酸化酶对髓袢升支粗短管周膜钾通道的作用研究 4、饮食钾摄入量改变及PTP对集合管管腔膜钾通道活性作用的研究 5、Ca2+对髓袢升支粗段管腔膜钾通道的影响及作用机制6、PGE2对髓袢升支粗短管周膜钾通道的影响及作用研究 7、腺苷对髓袢升支粗段管周膜钾通道的影响及作用机制 8、AA及代谢产物20-HETE对髓袢升支粗段管周膜氯离子通道的作用机制。从2000年开始,谷瑞民教授在美国、欧洲发表研究论文12篇,其中6篇第一作者的论文影响因子总和达34,分别发表在生理学界和肾脏研究领域的顶尖级杂志《普通生理学报》、《美国生理学报》及《世界肾脏》等刊物。2001年发表在美国生理学报的论文“The role of 20-HETE in mediating the effect of dietary K intake on the apical K channels in the mTAL”一文被sigma公司作为该公司每年产品目录20-HETE药物作用的佐证而收录。
谷瑞民教授目前作为项目主持人承担2项国家自然科学基金“肾脏髓袢升支粗段氯离子通道调控机制的研究”、“腺苷对髓袢升支粗段管周膜钾离子通道调控作用机制的研究”,1项国家教委博士点专项基金“花生四烯酸对肾脏髓袢升支粗段氯离子通道调控机制的研究”。
-
主页访问
1542
-
关注数
0
-
成果阅读
329
-
成果数
7
谷瑞民, RUIMIN GU, YUAN WEI, HOULI JIANG, MICHAEL BALAZY, AND WENHUI WANG
Am J Physiol Renal Physiol 280(2001)223-230,-0001,():
-1年11月30日
Gu, Ruimin, Yuan Wei, Houli Jiang, Michael Balazy, and Wenhui Wang. Role of 20-HETE in mediating the effect of dietary K intake on the apical K channels in the mTAL. Am J Physiol Renal Physiol 280: F223–F230, 2001.-We have used the patch-clamp technique to study the effect of dietary K intake on the apical K channels in the medullary thick ascending limb (mTAL) of rat kidneys. The channel activity, defined by the number of channels in a patch and the open probability (NPo), of the 30- and 70-pS K channels, was 0.18 and 0.11, respectively, in the mTAL from rats on a K-deficient diet. In contrast, NPo of the 30- and 70-pS K channels increased to 0.60 and 0.80, respectively, in the tubules from animals on a high-K diet. The concentration of 20-hydroxyeicosatetraenoic acid (20-HETE) measured with gas chromatography-mass spectrometry was 0.8 pg/mg protein in the mTAL from rats on a high-K diet and increased significantly to 4.6 pg/mg protein in the tubules from rats on a K-deficient diet. Addition of N-methylsulfonyl-12,12-dibromododec-11-enamide (DDMS) or 17-octadecynoic acid (17-ODYA), agents that inhibit the formation of 20-HETE, had no significant effect on the activity of the 30-pS K channels. However, DDMS/17-ODYA significantly increased the activity of the apical 70-pS K channel from 0.11 to 0.91 in the mTAL from rats on a K-deficient diet. In contrast, inhibition of the cytochrome P-450 metabolism of arachidonic acid increased NPo from 0.64 to 0.81 in the tubules from animals on a high-K diet. Furthermore, the sensitivity of the 70-pS K channel to 20-HETE was the same between rats on a high-K diet and on a K-deficient diet. Finally, the pretreatment of the tubules with DDMS increased NPo of the 70-pS K channels in the mTAL from rats on a K-deficient diet to 0.76. We conclude that an increase in 20-HETE production is involved in reducing the activity of the apical 70-pS K channels in the mTAL from rats on a K-deficient diet.
-
50浏览
-
0点赞
-
0收藏
-
0分享
-
86下载
-
0评论
-
引用
谷瑞民, RUI-MIN GU, , YUAN WEI, JOHN R. FALCK, U. MURALI KRISHNA, AND WEN-HUI WANGI
Am J Physiol Cell Physiol 281(2001)1188-1195,-0001,():
-1年11月30日
Gu, Rui-Min, Yuan Wei, John R. Falck, U. Murali Krishna, and Wen-Hui Wang. Effects of protein tyrosine kinase and protein tyrosine phosphatase on apical K1 channels in the TAL. Am J Physiol Cell Physiol 281: C1188-C1195, 2001.-We have previously demonstrated that the protein level of c-Src, a nonreceptor type of protein tyrosine kinase (PTK), was higher in the renal medulla from rats on a K-deficient (KD) diet than that in rats on a high-K (HK) diet (Wang WH, Lerea KM, Chan M, and Giebisch G. Am J Physiol Renal Physiol 278: F165–F171, 2000). We have now used the patch-clamp technique to investigate the role of PTK in regulating the apical K channels in the medullary thick ascending limb (mTAL) of the rat kidney. Inhibition of PTK with herbimycin A increased NPo, a product of channel number (N) and open probability (Po), of the 70-pS K channel from 0.12 to 0.42 in the mTAL only from rats on a KD diet but had no significant effect in tubules from animals on a HK diet. In contrast, herbimycin A did not affect the activity of the 30-pS K channel in the mTAL from rats on a KD diet. Moreover, addition of N-methylsulfonyl-12,12-dibromododec-11-enamide, an agent that inhibits the cytochrome P-450-dependent production of 20-hydroxyeicosatetraenoic acid, further increased NPo of the 70-pS K channel in the presence of herbimycin A. Furthermore, Western blot detected the presence of PTP-1D, a membrane-associated protein tyrosine phosphatase (PTP), in the renal outer medulla. Inhibition of PTP with phenylarsine oxide (PAO) decreased NPo of the 70-pS K channel in the mTAL from rats on a HK diet. However, PAO did not inhibit the activity of the 30-pS K channel in the mTAL. The effect of PAO on the 70-pS K channel was due to indirectly stimulating PTK because pretreatment of the mTAL with herbimycin A abolished the inhibitory effect of PAO. Finally, addition of exogenous c-Src reversibly blocked the activity of the 70-pS K channel in inside-out patches. We conclude that PTK and PTP have no effect on the low-conductance K channels in the mTAL and that PTK-induced tyrosine phosphorylation inhibits, whereas PTP-induced tyrosine dephosphorylation stimulates, the apical 70-pS K channel in the mTAL.
hypokalemia, hyperkalemia, c-Src, ROMK channel, herbimycin A, phenylarsine oxide
-
58浏览
-
0点赞
-
0收藏
-
0分享
-
76下载
-
0评论
-
引用
【期刊论文】Arachidonic acid inhibits K channels in basolateral membrane of the thick ascending limb
谷瑞民, RUI-MIN GU AND WEN-HUI WANG
Am J Phvsiol Renal Physiol 283(2002)407-414,-0001,():
-1年11月30日
Gu, Rui-Min, and Wen-Hui Wang. Arachidonic acid inhibits K channels in basolateral membrane of the thick ascending limb. Am J Physiol Renal Physiol 283: F407–F414, 2002. First published March 12, 2002; 10.1152/ajprenal. 00002.2002.-We have used the patch-clamp technique to study the effect of arachidonic acid (AA) on the basolateral K channels in the medullary thick ascending limb (mTAL) of rat kidney. An inwardly rectifying 50-pS K channel was identified in cell-attached and inside-out patches in the basolateral membrane of the mTAL. The channel open probability (Po) was 0.51 at the spontaneous cell membrane potential and decreased to 0.25 by 30mV hyperpolarization. The addition of 5M AA decreased channel activity, identified as NPo, from 0.58 to 0.08 in cell-attached patches. The effect of AA on the 50-pS K channel was specific because 10M cis-11,14,17-eicosatrienoic acid had no significant effect on channel activity. To determine whether the effect of AA was mediated by AA per se or by its metabolites, we examined the effect of AA on channel activity in the presence of indomethacin, an inhibitor of cyclooxygenase, or N-methylsulfonyl-12,12-dibromododec-11-enamide (DDMS), an inhibitor of cytochrome P-450 monooxygenase. Inhibition of cyclooxygenase increased channel activity from 0.54 to 0.9. However, indomethacin did not abolish the inhibitory effect of AA on the 50-pS K channel. In contrast, inhibition of cytochrome P-450 metabolism not only increased channel activity from 0.49 to 0.83 but also completely abolished the effect of AA. Moreover, addition of DDMS can reverse the inhibitory effect of AA on channel activity. The notion that the effect of AA was mediated by cytochrome P-450-dependent metabolites of AA is also supported by the observation that addition of 100 nM of 20-hydroxyeicosatetraenoic acid, a main metabolite of AA in the mTAL, can mimic the effect of AA. We conclude that AA inhibits the 50-pS K channel in the basolateral membrane of the mTAL and that the effect of AA is mainly mediated by cytochrome P-450-dependent metabolites of AA.
cyclooxygenase, cytochrome P-450-oxidation, 20-hydroxyeicosatetraenoic acid, basolateral K channel
-
39浏览
-
0点赞
-
0收藏
-
0分享
-
69下载
-
0评论
-
引用
谷瑞民, Hyacinth Sterling‡, Dao-Hong Lin‡, Rui-Min Gu‡, Ke Dong§¶, Steven C. Hebert§¶, and Wen-Hui Wang‡
THE JOURNAI OF BIOIOGICAI CHEMISTRY Vol. 277 No.64 (2002) 317-4323,-0001,():
-1年11月30日
We have previously shown that inhibiting proteintyrosine kinase increased whereas inhibiting proteintyrosine phosphatase (PTP) decreased renal outer medullary potassium channel 1 (ROMK1) channel activity (1). We have now used confocal microscopy, the patch clamp technique, and biotin labeling to further examine the role of tyrosine phosphorylation in regulating ROMK1 trafficking. Human embryonic kidney 293 cells were cotransfected with c-Src and green fluorescent protein-ROMK1, which has the same biophysical properties as those of ROMK1. Patch clamp studies have shown that phenylarsine oxide (PAO), an inhibitor of PTP, decreased the activity of ROMK1. Moreover, addition of PAO reduced the cell surface localization of green fluorescent protein-ROMK1 detected by confocal microscopy and diminished the surface ROMK1 density by 65% measured by biotin labeling. Also, PAO treatment significantly increased the phosphorylation of ROMK1. The notion that the effect of PAO is mediated by stimulating tyrosine phosphorylationinduced endocytosis of ROMK1 has also been supported by findings that mutating the tyrosine residue 337 of ROMK1 to alanine abolished the effect of PAO. Finally, the inhibitory effect of PAO on ROMK1 was completely blocked in the cells co-transfected with dominant negative dynamin (dynaminK44A). This indicates that the tyrosine phosphorylation-induced endocytosis of ROMK1 is dynamin-dependent. We conclude that inhibiting PTP increasesROMK1phosphorylationandresults inadynamindependent internalization of the channel.
-
49浏览
-
0点赞
-
0收藏
-
0分享
-
68下载
-
0评论
-
引用
谷瑞民, Yuan Wei, Peter Bloom, RuiMin Gu, and WenHui Wang
THE JOURNAI OF BIOIOGICAI CHEMISTRY Vol. 275 No.27 (2000) 0502-20507,-0001,():
-1年11月30日
Previous studies have demonstrated that an increase in the activity of protein-tyrosine kinase (PTK) is involved in the down-regulation of the activity of apical small conductance K1 (SK) channels in the cortical collecting duct (CCD) from rats on a K1-deficient diet (1). We used the patch clamp technique to investigate the role of protein-tyrosine phosphatase (PTP) in the regulation of the activity of SK channels in the CCD from rats on a high K1 diet. Western blot analysis indicated that PTP-1D is expressed in the renal cortex. Application of 1 mM phenylarsine oxide (PAO) or 1 mM benzylphosphonic acid, agents that inhibit PTP, reversibly reduced channel activity by 95%. Pretreatment of CCDs with PAO for 30 min decreased the mean NPo reversibly from control value 3.20 to 0.40. Addition of 1 mM herbimycin A, an inhibitor of PTK, had no significant effect on channel activity in the CCDs from rats on a high K1 diet. However, herbimycin A abolished the inhibitory effect of PAO, indicating that the effect of PAO is the result of interaction between PTK and PTP. Addition of brefeldin A, an agent that blocks protein trafficking from Golgi complex to the membrane, had no effect on channel activity. Moreover, application of colchicine, a microtubule inhibitor, or paclitaxel, a microtubule stabilizer, had no effect on channel activity. In contrast, PAO still reduced channel activity in the presence of brefeldin A, colchicine, or paclitaxel. Furthermore, the effect of PAO on channel activity was absent when the tubules were bathed in 16% sucrose-containing bath solution or treated with concanavalin A. We conclude that PTP is involved in the regulation of the activity of SK channels and that inhibition of PTP may facilitate the internalization of the SK channels.
-
41浏览
-
0点赞
-
0收藏
-
0分享
-
65下载
-
0评论
-
引用
【期刊论文】Regulation of ROMK1 Channels by Protein-tyrosine Kinase and-tyrosine Phosphatase*
谷瑞民, Zebunnessa Moral‡, Ke Dong§, Yuan Wei‡, Hyacinth Sterling‡, Huan Deng‡, Shariq Ali‡, RuiMin Gu‡, Xin-Yun Huang¶, Steven C. Hebert§, Gerhard Giebisch§, and Wen-Hui Wang‡¶
THE JOURNAI OF BIOIOGICAI CHEMISTRY Vol. 276 No.10 (2001) 7156-7163,-0001,():
-1年11月30日
We have used the two-electrode voltage clamp technique and the patch clamp technique to investigate the regulation of ROMK1 channels by protein-tyrosine phosphatase (PTP) and protein-tyrosine kinase (PTK) in oocytes coexpressing ROMK1 and cSrc. Western blot analysis detected the presence of the endogenous PTP-1D isoform in the oocytes. Addition of phenylarsine oxide (PAO), an inhibitor of PTP, reversibly reduced K1 current by 55% in oocytes coinjected with ROMK1 and cSrc. In contrast, PAO had no significant effect on K1 current in oocytes injected with ROMK1 alone. Moreover, application of herbimycin A, an inhibitor of PTK, increased K1 current by 120% and completely abolished the effect of PAO in oocytes coexpressing ROMK1 and cSrc. The effects of herbimycin A and PAO were absent in oocytes expressing the ROMK1 mutant R1Y337A in which the tyrosine residue at position 337 was mutated to alanine. However, addition of exogenous cSrc had no significant effect on the activity of ROMK1 channels in inside-out patches. Moreover, the effect of PAO was completely abolished by treatment of oocytes with 20% sucrose and 250 mg/ml concanavalin A, agents that inhibit the endocytosis of ROMK1 channels. Furthermore, the effect of herbimycin A is absent in the oocytes pretreated with either colchicine, an inhibitor of microtubules, or taxol, an agent that freezes microtubules. We conclude that PTP and PTK play an important role in regulating ROMK1 channels. Inhibiting PTP increases the internalization of ROMK1 channels, whereas blocking PTK stimulates the insertion of ROMK1 channels.
-
57浏览
-
0点赞
-
0收藏
-
0分享
-
40下载
-
0评论
-
引用
谷瑞民, YUAN WEI, PETER BLOOM, DAOHONG LIN, RUIMIN GU, AND WEN HUI WANG
Am J Physiol Renal Physiol 281(2001)206-212,-0001,():
-1年11月30日
Wei, Yuan, Peter Bloom, Daohong Lin, Ruimin Gu, and Wen Hui Wang. Effect of dietary K intake on apical small-conductance K channel in CCD: role of protein tyrosine kinase. Am J Physiol Renal Physiol 281: F206–F212, 2001.-We have used Western blot to examine the expression of cSrc protein tyrosine kinase (PTK) and protein tyrosine phosphatase (PTP)-1D in the renal cortex, and the patchclamp technique to determine the role of PTK in mediating the effect of dietary K intake on the small-conductance K (SK) channel in the cortical collecting duct (CCD). When rats were on a K-deficient (KD) diet for 1, 3, 5, and 7 days, the expression of cSrc increased by 40, 90, 140, and 135%, respectively. In contrast, the expression of cSrc in the renal cortex from rats on a high-K (HK) diet for 1, 2, and 3 days decreased by 40, 60, and 75%, respectively. However, the protein level of PTP-1D was not significantly changed by dietary K intake. The addition of 1 mM herbimycin A increased NPo, a product of channel number (N) and open probability (Po) in the CCD from rats on a normal diet or on a KD diet. The increase in NPo was 0.30 (normal), 0.45 (1-day KD), 0.65 (3-day KD), 1.55 (5-day KD), and 1.85 (7-day KD), respectively. Treatment of the CCD with herbimycin A from rats on a KD diet increased NPo per patch from the control value (0.7) to 1.4 (1-day KD), 1.6 (3-day KD), 2.6 (5-day KD), and 3.5 (7-day KD), respectively. In contrast, HK intake for as short as 1 day abolished the effect of herbimycin A. Furthermore, the expression of ROMK channels in the renal cortex was the same between rats on a KD diet or on a HK diet. Moreover, treatment with herbimycin A did not further increase NPo in the CCDs from rats on a HK diet. We conclude that dietary K intake plays a key role in regulating the activity of the SK channels and that PTK is involved in mediating the effect of the K intake on channel activity in the CCD.
hyperkalemia, hypokalemia, protein tyrosine phosphatase 1D, renal potassium secretion, cortical collecting duct
-
35浏览
-
0点赞
-
0收藏
-
0分享
-
34下载
-
0评论
-
引用