The purpose of the present study was to further study the characteristics and regulation of spontaneous transient outward currents(STOCs)in freshly isolated porcine coronary artery smooth muscle cells(ASMCs).STOCs wer...The purpose of the present study was to further study the characteristics and regulation of spontaneous transient outward currents(STOCs)in freshly isolated porcine coronary artery smooth muscle cells(ASMCs).STOCs were recorded using the perforated whole-cell patch-clamp configuration.STOCs were voltage-dependent and superimposed stochastically onto whole-cell Ca^(2+)-activated-K+(BKCa)currents.Charybdotoxin(ChTX,200 nmol/L),a selective blocker of BKCa channels,completely inhibited STOCs within 10 min.STOCs activity was greatly suppressed when extracellular Ca^(2+)concentration decreased from 1.8 mmol/L to 200 nmol/L,further removal of Ca^(2+)abolished STOCs activity.Ca^(2+)ionophore A23187(10μmol/L)increased STOCs activity significantly.Verapamil(20μmol/L)and CdCl2(200μmol/L),two kinds of organic L-type voltage-dependent Ca^(2+)channels(L-VDCCs)antagonists,had little effect on STOCs.In addition,the ryanodine receptors(RyRs)agonist caffeine(5 mmol/L)significantly activated STOCs.Application of ryanodine(50μmol/L)to block RyRs abolished STOCs,subsequent washout of ryanodine or application of caffeine failed to reproduce STOCs activity.Inhibition of inositol 1,4,5-trisphosphate receptors(IP_(3)Rs)by 2APB(40μmol/L)greatly suppressed the activity of STOCs,application of caffeine(5 mmol/L)in the presence of 2APB caused a burst of outward currents followed by inhibition of STOCs.These results suggest that STOCs in porcine coronary ASMCs are mediated by BKCa channels.Extracellular Ca^(2+)is essential for STOCs activity,while Ca^(2+)entry through L-VDCCs has little effect on STOCs.Intracellular Ca^(2+)release induced by RyRs is responsible for the regulation of STOCs,whereas IP_(3)Rs might also be involved.展开更多
The transmural heterogeneous changes of transient outward potassium currents (Ito) in rabbit hypertrophic cardiaomyocytes and the effects of long-term prophylactic treatment with volsartan were investigated. Rabbits w...The transmural heterogeneous changes of transient outward potassium currents (Ito) in rabbit hypertrophic cardiaomyocytes and the effects of long-term prophylactic treatment with volsartan were investigated. Rabbits were divided into hypertrophy group (left ventricular hypertrophy induced by partial ligation of abdominal aorta), vol-treated group (volsartan was administrated after the ligation), and control group (sham operated). Myocytes were isolated by a two-step enzymatical method. The sub-endocardial (Endo) and sub-epicardium (Epi) tissues were separated from midmyocardium (Mid) with a razor. Whole-cell patch-clamp technique was used to record potassium currents. The results showed that membrane capacitance was larger in hypertrophic cells than those in control and vol-treated cells (P<0.01 vs control cells, n=30). The densities of Ito in hypertrophic cells were reduced by sub-epicardium (Epi) (27.8±2.9) %, midmyocardium (Mid) (41.0±4.7) %, and sub-endocardium (Endo) (20.3±3.4) % compared with those in control cells. The decrease of Ito density was more pronounced in Mid than in Epi and Endo (P<0.01 vs Epi or Endo). There were no significant differences in Ito densities between vol-treated group and control group in three layers separately. In conclusion, volsartan can inhibit the transmural heterogeneous changes of Ito in left ventricular hypertrophic cardiomyocytes in rabbit.展开更多
The electrophysiological properties of potassium ion channels are regarded as a basic index for determining the functional differentiation of neural stem cells. In this study, neural stem cells from the hippocampus of...The electrophysiological properties of potassium ion channels are regarded as a basic index for determining the functional differentiation of neural stem cells. In this study, neural stem cells from the hippocampus of newborn rats were induced to differentiate with neurotrophic growth factor, and the electrophysiological properties of the voltage-gated potassium ion channels were observed. Immunofluorescence staining showed that the rapidly proliferating neural stem cells formed spheres in vitro that expressed high levels of nestin. The differentiated neurons were shown to express neuron-specific enolase. Flow cytometric analysis revealed that the neural stem cells were actively dividing and the percentage of cells in the S ＋ G2/M phase was high. However, the ratio of cells in the S ＋ G2/M phase decreased obviously as differentiation proceeded. Whole-cell patch-clamp re- cordings revealed apparent changes in potassium ion currents as the neurons differentiated. The potassium ion currents consisted of one transient outward potassium ion current and one delayed rectifier potassium ion current, which were blocked by 4-aminopyridine and tetraethylammonium, respectively. The experimental findings indicate that neural stem cells from newborn rat hippo- campus could be cultured and induced to differentiate into functional neurons under defined condi- tions in vitro. The differentiated neurons expressed two types of outward potassium ion cur＇ents similar to those of mature neurons in vivo.展开更多
Acutely isolated mouse hippocampal CA3 pyramidal neurons were exposed to 3 mT static magnetic field,and the characteristics of transient outward K+ channel were studied using the whole-cell patch-clamp technique.The e...Acutely isolated mouse hippocampal CA3 pyramidal neurons were exposed to 3 mT static magnetic field,and the characteristics of transient outward K+ channel were studied using the whole-cell patch-clamp technique.The experiment revealed that the amplitude of transient outward potassium channel current was reduced.The maximum activated current densities of control group and exposure group were 163.62±20.68 pA/pF and 98.74±16.57 pA/pF(n=12,P<0.01) respectively.The static magnetic field exposure affected the activation and inactivation process of transient outward potassium channel current.Due to the magnetic field exposure,the half-activation voltage of the activation curves changed from 5.59±1.96 mV to 27.87±7.24 mV(n=12,P<0.05) ,and the slope factor changed from 19.43±2.11 mV to 25.87±4.22 mV(n=12,P<0.05) .The half-inactivation voltage of the inactivation curves also changed from-56.09±0.89 mV to-57.16±1.10 mV(n=12,P>0.05) and the slope factor of the inactivation curves from 8.69±0.80 mV to 10.87±1.02 mV(n=12,P<0.05) .The results show that the static magnetic field can change the characteristics of transient outward K+ channel,and affect the physiological functions of neurons.展开更多
Background Some studies have confirmed that the right ventricular walls of most rodents, such as canines and humans, have evident transient outward potassium current (I to1) heterogeneity, and this heterogeneity i...Background Some studies have confirmed that the right ventricular walls of most rodents, such as canines and humans, have evident transient outward potassium current (I to1) heterogeneity, and this heterogeneity is closely related to J point elevation, J wave formation, and some ventricular tachycardias such as ventricular fibrillations caused by Brugada syndrome. This study is designed to investigate transmural electrical heterogeneity of the canine right ventricle during repolarization (phase 1) from the viewpoint of 4-aminopyridine sensitive and calcium-independent I to1. Methods Adult canine single right ventricular epicardial (Epi) cells, mid-myocardial (M) cells, and endocardial (Endo) cells were enzymatically dissociated. Whole cell voltage-clamp recordings were made to compare the I to1 values of the three cell types. Results At 37℃ and using 0.2 Hz and +70 mV depolarizing test potentials, the average peak I to1 values of Epi cells and M cells averaged (4070±1720) pA and (3540±1840) pA, respectively. The activated and inactivated Epi and M cells kinetic processes were in accordance with the Boltzmann distribution. Compared with I to1 in Epi cells and M cells, the average peak I to1 in Endo cells was very low, averaged (470±130) pA. Conclusions These results suggest that there are evident differences and potent gradients in I to1 between the three cardiac cell types, especially between Epi and Endo cells. These differences are among the prominent manifestations of right ventricular electrical heterogeneity, and may form an important ionic basis and prerequisite for some malignant arrhythmias in the right ventricle, including those arising from Brugada syndrome and other diseases.展开更多
目的:探讨参松养心胶囊对大鼠心室肌细胞L-型钙电流(Ica,L)和瞬时外向钾电流(Ito)的抑制作用。方法:酶解法分离大鼠心室肌细胞,使用参松养心溶液(0.5 mg/100 m L,0.5%)灌流心室肌细胞,利用全细胞膜片钳技术记录Ica,L和Ito,在细胞破膜后...目的:探讨参松养心胶囊对大鼠心室肌细胞L-型钙电流(Ica,L)和瞬时外向钾电流(Ito)的抑制作用。方法:酶解法分离大鼠心室肌细胞,使用参松养心溶液(0.5 mg/100 m L,0.5%)灌流心室肌细胞,利用全细胞膜片钳技术记录Ica,L和Ito,在细胞破膜后的20 min内记录所有数据,对每个细胞给药前后做自身对照,观察参松养心溶液对Ica,L和Ito通道电流的影响。结果:0.5%的参松养心溶液对Ica,L和Ito均有抑制作用,参松养心溶液对Ito的缓慢电流成分Itos无明显作用,但可以明显抑制Ito的快速电流成分Itof。在-10 m V测试电压下,在0.5%的参松养心溶液作用后,Ica,L的最大激活峰值电流密度从(-8.64+-2.43)p A/p F降为(-5.92+-2.15)p A/p F,抑制率为31.85%+10.25%,电流密度-电压(I-V)曲线上移;在-60 m V测试电压下,在0.5%的参松养心溶液作用后,Itof的最大激活峰值电流密度从(23.53+3.31)p A/p F降为(16.86+2.34)p A/p F,抑制率为(28.33+10.59)%,I-V曲线下移。参松养心溶液不改变Ica,L的通道动力学,但会使Ito通道失活后的恢复变慢。结论:参松养心溶液对Ica,L和Ito均有明显的抑制作用。展开更多
基金Supported by the National Natural Science Foundation of China (Grant No. 30370527)
文摘The purpose of the present study was to further study the characteristics and regulation of spontaneous transient outward currents(STOCs)in freshly isolated porcine coronary artery smooth muscle cells(ASMCs).STOCs were recorded using the perforated whole-cell patch-clamp configuration.STOCs were voltage-dependent and superimposed stochastically onto whole-cell Ca^(2+)-activated-K+(BKCa)currents.Charybdotoxin(ChTX,200 nmol/L),a selective blocker of BKCa channels,completely inhibited STOCs within 10 min.STOCs activity was greatly suppressed when extracellular Ca^(2+)concentration decreased from 1.8 mmol/L to 200 nmol/L,further removal of Ca^(2+)abolished STOCs activity.Ca^(2+)ionophore A23187(10μmol/L)increased STOCs activity significantly.Verapamil(20μmol/L)and CdCl2(200μmol/L),two kinds of organic L-type voltage-dependent Ca^(2+)channels(L-VDCCs)antagonists,had little effect on STOCs.In addition,the ryanodine receptors(RyRs)agonist caffeine(5 mmol/L)significantly activated STOCs.Application of ryanodine(50μmol/L)to block RyRs abolished STOCs,subsequent washout of ryanodine or application of caffeine failed to reproduce STOCs activity.Inhibition of inositol 1,4,5-trisphosphate receptors(IP_(3)Rs)by 2APB(40μmol/L)greatly suppressed the activity of STOCs,application of caffeine(5 mmol/L)in the presence of 2APB caused a burst of outward currents followed by inhibition of STOCs.These results suggest that STOCs in porcine coronary ASMCs are mediated by BKCa channels.Extracellular Ca^(2+)is essential for STOCs activity,while Ca^(2+)entry through L-VDCCs has little effect on STOCs.Intracellular Ca^(2+)release induced by RyRs is responsible for the regulation of STOCs,whereas IP_(3)Rs might also be involved.
文摘The transmural heterogeneous changes of transient outward potassium currents (Ito) in rabbit hypertrophic cardiaomyocytes and the effects of long-term prophylactic treatment with volsartan were investigated. Rabbits were divided into hypertrophy group (left ventricular hypertrophy induced by partial ligation of abdominal aorta), vol-treated group (volsartan was administrated after the ligation), and control group (sham operated). Myocytes were isolated by a two-step enzymatical method. The sub-endocardial (Endo) and sub-epicardium (Epi) tissues were separated from midmyocardium (Mid) with a razor. Whole-cell patch-clamp technique was used to record potassium currents. The results showed that membrane capacitance was larger in hypertrophic cells than those in control and vol-treated cells (P<0.01 vs control cells, n=30). The densities of Ito in hypertrophic cells were reduced by sub-epicardium (Epi) (27.8±2.9) %, midmyocardium (Mid) (41.0±4.7) %, and sub-endocardium (Endo) (20.3±3.4) % compared with those in control cells. The decrease of Ito density was more pronounced in Mid than in Epi and Endo (P<0.01 vs Epi or Endo). There were no significant differences in Ito densities between vol-treated group and control group in three layers separately. In conclusion, volsartan can inhibit the transmural heterogeneous changes of Ito in left ventricular hypertrophic cardiomyocytes in rabbit.
基金supported by the National Natural Science Foundation of China,No.31000514the Scientific Research Project for Talent with High Education of Xinxiang Medical University,No.2007502002
文摘The electrophysiological properties of potassium ion channels are regarded as a basic index for determining the functional differentiation of neural stem cells. In this study, neural stem cells from the hippocampus of newborn rats were induced to differentiate with neurotrophic growth factor, and the electrophysiological properties of the voltage-gated potassium ion channels were observed. Immunofluorescence staining showed that the rapidly proliferating neural stem cells formed spheres in vitro that expressed high levels of nestin. The differentiated neurons were shown to express neuron-specific enolase. Flow cytometric analysis revealed that the neural stem cells were actively dividing and the percentage of cells in the S ＋ G2/M phase was high. However, the ratio of cells in the S ＋ G2/M phase decreased obviously as differentiation proceeded. Whole-cell patch-clamp re- cordings revealed apparent changes in potassium ion currents as the neurons differentiated. The potassium ion currents consisted of one transient outward potassium ion current and one delayed rectifier potassium ion current, which were blocked by 4-aminopyridine and tetraethylammonium, respectively. The experimental findings indicate that neural stem cells from newborn rat hippo- campus could be cultured and induced to differentiate into functional neurons under defined condi- tions in vitro. The differentiated neurons expressed two types of outward potassium ion cur＇ents similar to those of mature neurons in vivo.
基金Supported by National Natural Science Foundation of China(No. 60674111)
文摘Acutely isolated mouse hippocampal CA3 pyramidal neurons were exposed to 3 mT static magnetic field,and the characteristics of transient outward K+ channel were studied using the whole-cell patch-clamp technique.The experiment revealed that the amplitude of transient outward potassium channel current was reduced.The maximum activated current densities of control group and exposure group were 163.62±20.68 pA/pF and 98.74±16.57 pA/pF(n=12,P<0.01) respectively.The static magnetic field exposure affected the activation and inactivation process of transient outward potassium channel current.Due to the magnetic field exposure,the half-activation voltage of the activation curves changed from 5.59±1.96 mV to 27.87±7.24 mV(n=12,P<0.05) ,and the slope factor changed from 19.43±2.11 mV to 25.87±4.22 mV(n=12,P<0.05) .The half-inactivation voltage of the inactivation curves also changed from-56.09±0.89 mV to-57.16±1.10 mV(n=12,P>0.05) and the slope factor of the inactivation curves from 8.69±0.80 mV to 10.87±1.02 mV(n=12,P<0.05) .The results show that the static magnetic field can change the characteristics of transient outward K+ channel,and affect the physiological functions of neurons.
文摘Background Some studies have confirmed that the right ventricular walls of most rodents, such as canines and humans, have evident transient outward potassium current (I to1) heterogeneity, and this heterogeneity is closely related to J point elevation, J wave formation, and some ventricular tachycardias such as ventricular fibrillations caused by Brugada syndrome. This study is designed to investigate transmural electrical heterogeneity of the canine right ventricle during repolarization (phase 1) from the viewpoint of 4-aminopyridine sensitive and calcium-independent I to1. Methods Adult canine single right ventricular epicardial (Epi) cells, mid-myocardial (M) cells, and endocardial (Endo) cells were enzymatically dissociated. Whole cell voltage-clamp recordings were made to compare the I to1 values of the three cell types. Results At 37℃ and using 0.2 Hz and +70 mV depolarizing test potentials, the average peak I to1 values of Epi cells and M cells averaged (4070±1720) pA and (3540±1840) pA, respectively. The activated and inactivated Epi and M cells kinetic processes were in accordance with the Boltzmann distribution. Compared with I to1 in Epi cells and M cells, the average peak I to1 in Endo cells was very low, averaged (470±130) pA. Conclusions These results suggest that there are evident differences and potent gradients in I to1 between the three cardiac cell types, especially between Epi and Endo cells. These differences are among the prominent manifestations of right ventricular electrical heterogeneity, and may form an important ionic basis and prerequisite for some malignant arrhythmias in the right ventricle, including those arising from Brugada syndrome and other diseases.
