Area of Toxicology, Faculty of Pharmacy, University of Seville, Seville, Spain.
Aquat Toxicol. 2011 Jan 17;101(1):109-16. doi: 10.1016/j.aquatox.2010.09.009. Epub 2010 Sep 25.
The cyanobacteria Cylindrospermopsis raciborskii is considered a threat to aquatic organisms due to the production of the toxin cylindrospermopsin (CYN). Despite the numerous reports evidencing the toxic effects of C. raciborskii cells and CYN in different species, not much is known regarding the toxicity mechanisms associated with this toxin and the cyanobacteria. In this work, a proteomics approach based in the two-dimensional gel electrophoresis and mass spectrometry was used to study the effects of the exposure of two bivalve species, Mytilus galloprovincialis and Corbicula fluminea, to CYN producing (CYN+) and non-producing (CYN-) C. raciborskii cells. Additionally the activities of glutathione S-transferase (GST) and glutathione peroxidase (GPx) were determined. Alterations in actin and tubulin isoforms were detected in gills of both bivalve species and digestive gland of M. galloprovincialis when exposed to CYN- and CYN+ cells. Moreover, GST and GPx activities changed in gills and digestive tract of bivalves exposed to both C. raciborskii freeze dried cells, in comparison to control animals exposed to the green alga Chlorella vulgaris. These results suggest the induction of physiological stress and tissue injury in bivalves by C. raciborskii. This condition is supported by the changes observed in GPx and GST activities which indicate alterations in the oxidative stress defense mechanisms. The results also evidence the capacity of CYN non-producing C. raciborskii to induce biochemical responses and therefore its toxicity potential to bivalves. The heat shock protein 60 (HSP60), extrapallial (EP) fluid protein and triosephosphate isomerase homologous proteins from gills of M. galloprovincialis were down-regulated specifically with the presence of CYN+ C. raciborskii cells. The presence of CYN may lead to additional toxic effects in M. galloprovincialis. This work demonstrates that proteomics is a powerful approach to characterize the biochemical effects of C. raciborskii and to investigate the physiological condition of the exposed organisms.
集胞藻（Cylindrospermopsis raciborskii）被认为是水生生物的威胁，因为它会产生毒素微囊藻毒素（CYN）。尽管有大量报道证实了 C. raciborskii 细胞和 CYN 在不同物种中的毒性作用，但对于与这种毒素和蓝藻相关的毒性机制知之甚少。在这项工作中，采用基于二维凝胶电泳和质谱的蛋白质组学方法研究了两种双壳类动物贻贝（Mytilus galloprovincialis）和河蚬（Corbicula fluminea）暴露于产生 CYN（CYN+）和不产生 CYN（CYN-）的 C. raciborskii 细胞后的影响。此外，还测定了谷胱甘肽 S-转移酶（GST）和谷胱甘肽过氧化物酶（GPx）的活性。当暴露于 CYN-和 CYN+细胞时，两种双壳类动物的鳃和贻贝的消化腺中都检测到肌动蛋白和微管蛋白同工型的改变。此外，与暴露于绿藻小球藻的对照组动物相比，暴露于 C. raciborskii 冻干细胞的双壳类动物的鳃和消化道中的 GST 和 GPx 活性发生了变化。这些结果表明，C. raciborskii 会诱导双壳类动物产生生理应激和组织损伤。这种情况得到了 GST 和 GPx 活性变化的支持，这表明氧化应激防御机制发生了改变。结果还证明了不产生 CYN 的 C. raciborskii 具有诱导生化反应的能力，因此对双壳类动物具有潜在的毒性。来自贻贝鳃的热休克蛋白 60（HSP60）、外套膜（EP）液蛋白和磷酸丙糖异构酶同源蛋白在 CYN+ C. raciborskii 细胞存在的情况下特异性下调。CYN 的存在可能会导致贻贝产生额外的毒性作用。这项工作表明，蛋白质组学是一种强大的方法，可以用于描述 C. raciborskii 的生化效应，并研究暴露生物的生理状况。
