Trela Zenon, Burdach Zbigniew, Siemieniuk Agnieszka, Przestalski Stanisław, Karcz Waldemar
Department of Physics and Biophysics, Wrocław University of Environmental and Life Sciences, Norwida 25, PL-50-375, Wrocław, Poland.
Department of Plant Physiology, Faculty of Biology and Environmental Protection, University of Silesia, Jagiellońska 28, PL-40-032, Katowice, Poland.
PLoS One. 2015 Aug 28;10(8):e0136346. doi: 10.1371/journal.pone.0136346. eCollection 2015.
In the present study, patch-clamp techniques have been used to investigate the effect of trimethyltin chloride (Met3SnCl) on the slow vacuolar (SV) channels in vacuoles from red beet (Beta vulgaris L.) taproots. Activity of SV channels has been measured in whole-vacuole and cytosolic side-out patch configurations. It was found that addition of trimethyltin chloride to the bath solution suppressed, in a concentration-dependent manner, SV currents in red beet vacuoles. The time constant, τ, increased significantly in the presence of the organotin. When single channel activity was analyzed, only little channel activity could be recorded at 100 μM Met3SnCl. Trimethyltin chloride added to the bath medium significantly decreased (by ca. threefold at 100 μM Met3SnCl and at 100 mV voltage, as compared to the control medium) the open probability of single channels. Single channel recordings obtained in the presence and absence of trimethyltin chloride showed that the organotin only slightly (by <10%) decreased the unitary conductance of single channels. It was also found that Met3SnCl significantly diminished the number of SV channel openings, whereas it did not change the opening times of the channels. Taking into account the above and the fact that under the here applied experimental conditions (pH = 7.5) Met3SnCl is a non-dissociated (more lipophilic) compound, we suggest that the suppression of SV currents observed in the presence of the organotin results probably from its hydrophobic properties allowing this compound to translocate near the selectivity filter of the channel.
在本研究中,采用膜片钳技术研究了三甲基氯化锡(Met3SnCl)对红甜菜(Beta vulgaris L.)主根液泡中慢液泡(SV)通道的影响。在全液泡和胞质侧向外膜片配置下测量了SV通道的活性。发现向浴液中添加三甲基氯化锡以浓度依赖性方式抑制了红甜菜液泡中的SV电流。在有机锡存在下,时间常数τ显著增加。当分析单通道活性时,在100 μM Met3SnCl时只能记录到很少的通道活性。添加到浴液介质中的三甲基氯化锡显著降低了单通道的开放概率(在100 μM Met3SnCl和100 mV电压下,与对照介质相比约降低了三倍)。在有和没有三甲基氯化锡的情况下获得的单通道记录表明,有机锡仅略微(<10%)降低了单通道的单位电导。还发现Met3SnCl显著减少了SV通道的开放次数,而没有改变通道的开放时间。考虑到上述情况以及在本文应用的实验条件(pH = 7.5)下Met3SnCl是一种未解离(更具亲脂性)的化合物这一事实,我们认为在有机锡存在下观察到的SV电流抑制可能是由于其疏水特性使该化合物能够转运到通道选择性过滤器附近。