CHANGES IN GAP JUNCTION PERMEABILITY, GAP JUNCTION NUMBER, AND CONNEXIN43 EXPRESSION IN LINDANE-TREATED RAT-LIVER EPITHELIAL-CELLS
CHANGES IN GAP JUNCTION PERMEABILITY, GAP JUNCTION NUMBER, AND CONNEXIN43 EXPRESSION IN LINDANE-TREATED RAT-LIVER EPITHELIAL-CELLS
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DOI：
10.1006/taap.1995.1011
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发表时间：
1995-01-01
影响因子：
3.8
通讯作者：
RUCH, RJ
中科院分区：
文献类型：
--
作者：
GUAN, XJ;BONNEY, WJ;RUCH, RJ
农药林丹（γ -六氯环己烷）是一种哺乳动物神经毒素和肝癌致癌物。林丹具有抑制间隙连接细胞间通讯（GJIC）的作用，这可能与林丹的毒性有关。研究林丹对WB-F344大鼠肝上皮细胞GJIC的抑制机制，以确定是否与间隙连接通透性、间隙连接数量和/或间隙连接蛋白（connexin43）表达的改变有关。荧光路西法黄CH染料显微注射（染料偶联）监测GJIC。使用抗connexin43单克隆抗体间接免疫染色间隙连接后，目视定量间隙连接数。分别用Northern blotting和Western blotting检测Connexin43 mRNA和蛋白水平。林丹（50 μ M）短期处理（10-30 min）导致染料偶联迅速（10 min内）几乎完全丧失，但间隙连接数和connexin43 mRNA或蛋白水平未发生变化。中期处理（1-4小时）导致染料偶联、间隙连接和磷酸化的connexin43蛋白的丧失。长期处理（1-14天）导致染料偶联、总connexin43蛋白和connexin43 mRNA减少。核运行试验表明，林丹非特异性地降低了connexin43基因的转录。这些数据表明，间隙结通透性、数量和表达的减少可能与GJIC的抑制有关，这取决于农药处理时间。(C) 1995学术出版社，Inc。
The pesticide lindane (gamma-hexachlorocyclohexane) is a mammalian neurotoxin and hepatocarcinogen. Lindane can inhibit gap junctional intercellular communication (GJIC) and this effect may contribute to its toxic properties. The mechanism of inhibition of GJIC by lindane in WB-F344 rat liver epithelial cells was studied to determine if altered gap junction permeability, gap junction number, and/or gap junction protein (connexin43) expression were involved. GJIC was monitored by fluorescent Lucifer Yellow CH dye microinjection (dye-coupling). Gap junction number was quantified visually after indirect immunostaining of gap junctions using an anti-connexin43 monoclonal antibody. Connexin43 mRNA and protein levels were determined by Northern and Western blotting, respectively. Short-term treatment (10-30 min) with lindane (50 mu M) resulted in the rapid (within 10 min), nearly complete loss of dye-coupling but no changes in gap junction number or connexin43 mRNA or protein levels. Medium-term treatment (1-4 hr) resulted in the loss of dye-coupling, gap junctions, and phosphorylated connexin43 proteins. Long-term treatment (1-14 days) led to reductions in dye-coupling, total connexin43 protein, and connexin43 mRNA. Nuclear run-on assays indicated that transcription of the connexin43 gene was reduced nonspecifically by lindane. These data indicate that reductions in gap junction permeability, number, and expression may be involved in the inhibition of GJIC depending on pesticide treatment duration. (C) 1995 Academic Press, Inc.