Aim: To record the single-channel currents and characterize the electrophysiological properties of the Cl^- channels inhuman sperm membrane. Methods: The membrane proteins extracted from the human sperm were reassembl...Aim: To record the single-channel currents and characterize the electrophysiological properties of the Cl^- channels inhuman sperm membrane. Methods: The membrane proteins extracted from the human sperm were reassembled intoliposome bilayer, and the liposomes were fused into giant liposomes with a diameter more than 10μm by dehydration-rehydration procedure. The giant liposomes were used to study the Cl^- channel activities by patch-clamp technique.Results: By patch clamping the giant liposome in an asymmetric NMDG (N-methyl-D-glucamine)-Cl (bath 100//pipette 200 mmol/L) solution system, three kinds of single-channel events with unit conductances of (74.1 ± 8.3) pS,(117.0±5.7) pS and (144.7±4.5) pS, respectively, were detected. Their activities were voltage-dependent and allwere blocked by SITS (4-acetamido-4'-isothiocyanato-stilbene-2', 2'-disulfonic acid) in a concentration-dependentmanner. By constructing the open and close dwell time distribution histograms and then fitting them with exponentialfunction, two time constants were obtained in both the open and the close states. The burst activity and conductancesubstate of the channels were observed. Conclusion; There exist three kinds of Cl^- channels with different conduc-tance in human sperm membrane at least. (Asian J Androl 2001 Sep; 3: 185 - 191)展开更多
Previous data showed that a Na+-transmem-brane flux was accompanied with acrosome reaction of sperm. However, the electrophysiological recording and characterization of Na+ current in human sperm membrane have not bee...Previous data showed that a Na+-transmem-brane flux was accompanied with acrosome reaction of sperm. However, the electrophysiological recording and characterization of Na+ current in human sperm membrane have not been yet reported. In the present investigation, membrane proteins extracted from human sperms were reassembled into liposome bilayer, and then the liposomes were fused by dehydration-rehydration into giant liposomes with the diameter of more than 10μm. By patch clamping the giant liposomes two kinds of single channel currents were recorded in a NaCl solution system. Both of them were Na+-carried, TTX-sensitive and strongly rectifying, but with different unit conductance and open probability. Moreover, bursting activity and channel-substates as well as two open time constants were observed in the larger channel.展开更多
文摘Aim: To record the single-channel currents and characterize the electrophysiological properties of the Cl^- channels inhuman sperm membrane. Methods: The membrane proteins extracted from the human sperm were reassembled intoliposome bilayer, and the liposomes were fused into giant liposomes with a diameter more than 10μm by dehydration-rehydration procedure. The giant liposomes were used to study the Cl^- channel activities by patch-clamp technique.Results: By patch clamping the giant liposome in an asymmetric NMDG (N-methyl-D-glucamine)-Cl (bath 100//pipette 200 mmol/L) solution system, three kinds of single-channel events with unit conductances of (74.1 ± 8.3) pS,(117.0±5.7) pS and (144.7±4.5) pS, respectively, were detected. Their activities were voltage-dependent and allwere blocked by SITS (4-acetamido-4'-isothiocyanato-stilbene-2', 2'-disulfonic acid) in a concentration-dependentmanner. By constructing the open and close dwell time distribution histograms and then fitting them with exponentialfunction, two time constants were obtained in both the open and the close states. The burst activity and conductancesubstate of the channels were observed. Conclusion; There exist three kinds of Cl^- channels with different conduc-tance in human sperm membrane at least. (Asian J Androl 2001 Sep; 3: 185 - 191)
基金National Natural Science Foundation of China(61025021,60936002,51072089,51372130)National Key Project of Science and Technology(2011ZX02403-002)Natural Science Foundation of Beijing(NSF 3111002)
基金This work was supported by the National Natural Science Foundation of China (Grant No. 98700197).
文摘Previous data showed that a Na+-transmem-brane flux was accompanied with acrosome reaction of sperm. However, the electrophysiological recording and characterization of Na+ current in human sperm membrane have not been yet reported. In the present investigation, membrane proteins extracted from human sperms were reassembled into liposome bilayer, and then the liposomes were fused by dehydration-rehydration into giant liposomes with the diameter of more than 10μm. By patch clamping the giant liposomes two kinds of single channel currents were recorded in a NaCl solution system. Both of them were Na+-carried, TTX-sensitive and strongly rectifying, but with different unit conductance and open probability. Moreover, bursting activity and channel-substates as well as two open time constants were observed in the larger channel.
