A new method for in vitro detection of microbially produced mitochondrial toxins
A new method for in vitro detection of microbially produced mitochondrial toxins
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DOI：
10.1016/s0887-2333(03)00097-3
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发表时间：
2003-10-01
影响因子：
3.2
通讯作者：
Salkinoja-Salonen, MS
中科院分区：
文献类型：
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作者：
Hoornstra, D;Andersson, MA;Salkinoja-Salonen, MS
精子运动抑制试验，早期显示出有价值的检测食物中毒的非蛋白毒素的芽孢杆菌物种被开发成一种检测线粒体损伤毒素有用的特异性。这是通过在质膜渗透性屏障保持完整的条件下评估线粒体内膜跨膜电位Δ Psi（m）的耗散来完成的。Δ Psi（m）估计为暴露精子的线粒体鞘中橙子JC-1荧光的强度。通过观察碘化丙啶从细胞质中的排除来评估相同细胞的质膜完整性。三种类型的线粒体毒性反应微生物产生的生物活性物质被确认。短杆菌肽的线粒体毒性（A.，B、C、D）、尼日利亚菌素、盐霉素、甲基盐霉素、莫能菌素、卡西霉素和抗霉素A的特征在于JC-1荧光在线粒体中逐渐褪色。Deltapsi（m）被cereulide、缬氨霉素和恩镰孢菌素（A，A（1），B，B-1）耗散，可见线粒体JC-1荧光的点状猝灭。此外，这些物质引起质膜超极化。寡霉素（A、B、C）、离子霉素和星形孢菌素抑制精子运动，但Deltapsi（m）精子活力得以完全保存。表面活性素和地衣多糖A在质膜也被破坏的浓度下引起线粒体损伤。(C)2003爱思唯尔有限公司。保留所有权利。
Sperm motility inhibition assay, earlier shown valuable for the detection of food poisoning non-protein toxins of Bacillus species was developed into an assay useful for specific detection of mitochondria damaging toxins. This was done by assessing the dissipation of the mitochondrial inner membrane transmembrane potential, Deltapsi(m) under conditions where the plasma membrane permeability barrier remained intact. The Deltapsi(m) was estimated as the intensity of orange JC-1 fluorescence in the mitochondrial sheath of the exposed spermatozoa. The plasma membrane integrity of the same cells was assessed by observing the exclusion of propidium iodide from the cytoplasm. Three types of mitochondrial toxic responses to microbially made bioactive substances were recognised. Mitochondrial toxicity by gramicidin (A., B, C, D), nigericin, salinomycin, narasin, monensin, calcimycin and antimycin A was characterised by gradual fading of the JC-1 fluorescence in the mitochondria. Dissipation of the Deltapsi(m) by cereulide, valinomycin and enniatin (A, A(1), B, B-1) was visible as spotwise quenching of the mitochondrial JC-1 fluorescence. In addition these substances caused hyperpolarisation of the plasma membrane. Oligomycin (A, B, C), ionomycin and staurosporine inhibited the spermatozoan motility, but Deltapsi(m) was fully preserved. Surfactin and lichenysin A caused mitochondrial damage at concentrations where the plasma membrane was also damaged. (C) 2003 Elsevier Ltd. All rights reserved.