Distinctive gap junction channel types connect WB cells, a clonal cell line derived from rat liver.
Distinctive gap junction channel types connect WB cells, a clonal cell line derived from rat liver.
复制标题
独特的间隙连接通道类型连接 WB 细胞，这是一种源自大鼠肝脏的克隆细胞系。
DOI：
10.1152/ajpcell.1991.260.3.c513
复制
发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Meda,P
中科院分区：
文献类型：
--
作者：
Spray,DC;Chanson,M;Moreno,AP;Dermietzel,R;Meda,P
间隙连接，染料耦合，和交界处的电导进行了研究，在细胞系（WB），是来自大鼠肝脏，并显示类似于“椭圆形”细胞的表型。在冷冻断裂复制品，两个独特的颗粒大小检测间隙连接斑块。免疫细胞化学研究表明，点状染色在膜附着使用抗体连接蛋白43和脑间隙连接相关抗原（34 kDa）。使用针对大鼠肝脏间隙连接蛋白（连接蛋白32和26）制备的抗体未观察到染色。WB电池对被电偶联和染料偶联。细胞对之间的结电导（gj）平均约为10 nS;偶尔，gj足够低，可以检测到单位结电导（γ j）。使用含CsCl的电极溶液，记录了不同的γ j值：约20-30 pS，约80-90 pS，以及其他大小的总和。最大的伽马j事件显然是由于随机重合的开口或较小的通道关闭。几种处理降低了gj。γ j的频率分布不变的2 mM氟烷或3.5庚醇，但中间和最大的事件的大小略有减少100 nM佛波酯，和最大的事件的相对频率增加10 μ M戊二醛。我们的结论是，独特的伽马j值代表两种不同类型的间隙连接通道，而不是一个单一的通道类型的子状态的开口和关闭，这些单一的电导可能对应于双重免疫反应性和冷冻断裂中看到的两个颗粒大小。
Gap junctions, dye coupling, and junctional conductance were studied in a cell line (WB) that is derived from rat liver and displays a phenotype similar to “oval” cells. In freeze-fracture replicas, two distinctive particle sizes were detected in gap junctional plaques. Immunocytochemical studies indicated punctate staining at membrane appositions using antibodies to connexin 43 and to a brain gap junction-associated antigen (34 kDa). No staining was observed using antibodies prepared against rat liver gap junction proteins (connexins 32 and 26). Pairs of WB cells were electrically and dye coupled. Junctional conductance (gj) between cell pairs averaged approximately 10 nS; occasionally, gj was low enough that unitary junctional conductances (gamma j) could be detected. Using a CsCl-containing electrode solution, distinctive gamma j values were recorded: approximately 20-30 pS, approximately 80-90 pS, and the sum of the other sizes. The largest gamma j events were apparently due to random coincident openings or closures of the smaller channels. Several treatments reduced gj. Frequency distributions of gamma j were unaltered by 2 mM halothane or 3.5 heptanol, but the sizes of intermediate and largest events were reduced slightly by 100 nM phorbol ester, and the relative frequency of the largest events was increased by 10 microM glutaraldehyde. We conclude that the distinctive gamma j values represent openings and closures of two distinct types of gap junction channels rather than substates of a single channel type; these unitary conductances may correspond to the dual immunoreactivity and to the two particle sizes seen in freeze fracture.