The signaling mechanisms of long distance intercellular calcium waves (far waves) in cultured human uterine myocytes.
The signaling mechanisms of long distance intercellular calcium waves (far waves) in cultured human uterine myocytes.
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培养的人子宫肌细胞中长距离细胞间钙波（远波）的信号机制。
DOI：
10.1023/a:1022052910585
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发表时间：
2002
影响因子：
2.7
通讯作者：
Zhang,Peisheng
中科院分区：
文献类型：
--
作者：
Young,RogerC;Schumann,Ralph;Zhang,Peisheng
培养的人心肌细胞显示出细胞间钙波，其传播距离超过100 μm（“远波”）。本文研究了远波传播的机理。从足月妊娠妇女的子宫肌层活检开始培养细胞系。钙绿色-1被用作细胞内游离钙的荧光探针。以0.83帧/秒的帧速率进行连续成像。细胞间钙波是通过无创伤地将小滴矿物油施加到单层的表面上来机械启动的。使用标准化网格对每个细胞间钙波进行评分，并根据波传播的距离和每个区域内观察到的荧光强度分配点数。在间隙连接和连接蛋白半通道（辛醇）、ATP（腺苷三磷酸双磷酸酶和MDL 12330 A）、前列腺素（吲哚美辛，高浓度镧）、前列腺素转运蛋白、PGT（DIDS）和跨膜钙通量（低浓度镧）的抑制剂存在下进行实验。辛醇，腺苷三磷酸双磷酸酶和MDL 12330 A未能修改远波，表明间隙连接，连接蛋白半通道和ATP不参与旁分泌机制。30、100和300 μM吲哚美辛以剂量依赖性方式将远波评分降至0，表明前列腺素在该机制中起关键作用。DIDS降低了远波评分，但没有完全抑制波传播，表明存在PGT依赖性和非依赖性成分的机制。0.1 mM的镧没有影响，但在1 mM时，降低了远波评分。这些结果与PGF 2 α和/或PGE 2是PGT依赖性组分的信号分子一致。两者合计，这些数据表明，长距离细胞间钙波在培养的人心肌细胞利用旁分泌信号机制，但与一个以上的细胞外信号化合物。
Cultured human myocytes exhibit intercellular calcium waves that travel farther than 100 μm (‘far waves’). This work investigates the mechanism of far wave propagation. Culture lines were initiated from myometrial biopsies of term pregnant women. Calcium green-1 was used as a fluorescence probe for intracellular free calcium. Serial imaging was performed at a frame rate of 0.83 frames/s. Intercellular calcium waves were mechanically initiated by atraumatically applying small drops of mineral oil onto the surface of the monolayer. Each intercellular calcium wave was scored using a standardized grid, and points were assigned depending upon the distance the wave traveled and the fluorescent intensity observed within each region. Experiments were performed in the presence of inhibitors of gap junctions and connexin hemichannels (octanol), ATP (apyrase and MDL 12330 A), prostaglandins (indomethacin, high concentrations of lanthanum), the prostaglandin transporter, PGT (DIDS), and transmembrane calcium flux (low concentrations of lanthanum). Octanol, apyrase and MDL 12330 A failed to modify the far waves, indicating gap junctions, connexin hemichannels and ATP do not participate in the paracrine mechanism. Indomethacin at 30, 100 and 300 μM, in a dose dependent manner, reduced the far wave score to 0, suggesting a prostaglandin was critically involved in the mechanism. DIDS reduced the far wave score, but did not fully inhibit wave propagation, suggesting the presence of PGT-dependent and -independent components to the mechanism. Lanthanum at 0.1 mM had no effect, but at 1 mM, reduced the far wave score. These results are consistent with PGF2αand/or PGE2being the signal molecule for the PGT-dependent component. Taken together, these data indicate that long distance intercellular calcium waves in cultured human myocytes utilizes a paracrine signaling mechanism, but with more than one extracellular signaling compound.
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DOI：
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发表时间：
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期刊：
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作者：
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TRIANDIS, HC;LISANSKY, J;BETANCOURT, H
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