A CHLORIDE CHANNEL FROM HUMAN PLACENTA RECONSTITUTED INTO GIANT LIPOSOMES

被引:27
作者
RIQUELME, G [1 ]
STUTZIN, A [1 ]
BARROS, LF [1 ]
LIBERONA, JL [1 ]
机构
[1] UNIV CHILE,SCH MED,DEPT EXPTL MED,SANTIAGO 7,CHILE
关键词
CHLORIDE CHANNEL; HUMAN PLACENTA; ION CHANNEL RECONSTITUTION; PATCH-CLAMP METHOD;
D O I
10.1016/0002-9378(95)90332-1
中图分类号
R71 [妇产科学];
学科分类号
100211 ;
摘要
OBJECTIVE: Ion channels play important roles in epithelial transport, but they are difficult to access for conventional electrophysiologic studies in intact placenta. The purpose of this work was to explore the suitability of purified trophoblast plasma membrane as a source of ion channels for reconstitution in artificial lipid membranes. STUDY DESIGN: Human placental brush border membranes were purified by differential and gradient centrifugation and fused with small liposomes. Giant liposomes were then generated by a cycle of dehydration and rehydration. These giant liposomes are suitable for electrophysiologic studies and were probed for the presence of active ion channels by the patch-clamp method. RESULTS: The results reported here indicate the presence of a high conductance chloride channel showing some similarities with ''maxi'' chloride channels described in secreting and absorbing epithelia. The channel had a slight outward rectification with conductances of 232 and 300 pS at negative and positive potentials, respectively. CONCLUSIONS: For the first time successful reconstitution of a human placental ion channel is achieved in a system suited for electrophysiologic studies. The chloride channel described might play a role in transplacental transport.
引用
收藏
页码:733 / 738
页数:6
相关论文
共 28 条
[1]  
BETTENDORFF L, 1993, J MEMBRANE BIOL, V136, P281
[2]   SINGLE VOLTAGE-DEPENDENT CHLORIDE-SELECTIVE CHANNELS OF LARGE CONDUCTANCE IN CULTURED RAT MUSCLE [J].
BLATZ, AL ;
MAGLEBY, KL .
BIOPHYSICAL JOURNAL, 1983, 43 (02) :237-241
[3]  
BOYD CAR, 1994, JPN J PHYSIOL, V44, pS273
[4]   CHLORIDE CHANNELS OF HIGH CONDUCTANCE IN THE MICROVILLOUS MEMBRANE OF TERM HUMAN PLACENTA [J].
BROWN, PD ;
GREENWOOD, SL ;
ROBINSON, J ;
BOYD, RDH .
PLACENTA, 1993, 14 (01) :103-115
[5]   CHLORIDE CONDUCTANCE IN MEMBRANE-VESICLES FROM HUMAN-PLACENTA USING A FLUORESCENT-PROBE - IMPLICATIONS FOR CYSTIC-FIBROSIS [J].
DECHECCHI, MC ;
CABRINI, G .
BIOCHIMICA ET BIOPHYSICA ACTA, 1988, 945 (02) :113-120
[6]  
FALLER D, 1992, J MEMBRANE BIOL, V130, P227
[7]   CHLORIDE CHANNELS OF BIOLOGICAL-MEMBRANES [J].
FRANCIOLINI, F ;
PETRIS, A .
BIOCHIMICA ET BIOPHYSICA ACTA, 1990, 1031 (02) :247-259
[8]   PURIFICATION AND NA+ UPTAKE BY HUMAN PLACENTAL MICROVILLUS MEMBRANE-VESICLES PREPARED BY 3 DIFFERENT METHODS [J].
GLAZIER, JD ;
JONES, CJP ;
SIBLEY, CP .
BIOCHIMICA ET BIOPHYSICA ACTA, 1988, 945 (02) :127-134
[9]   CHLORIDE CHANNELS IN EPITHELIA [J].
GOGELEIN, H .
BIOCHIMICA ET BIOPHYSICA ACTA, 1988, 947 (03) :521-547
[10]   HIGH CONDUCTANCE ANION-SELECTIVE CHANNELS IN RAT CULTURED SCHWANN-CELLS [J].
GRAY, PTA ;
BEVAN, S ;
RITCHIE, JM .
PROCEEDINGS OF THE ROYAL SOCIETY SERIES B-BIOLOGICAL SCIENCES, 1984, 221 (1225) :395-409