Cañete Elisabeth, Diogène Jorge
Centre d' Aqüicultura, IRTA, Ctra Poble Nou s/n Km 5.5, 43540 Sant Carles de la Rapita, Tarragona, Spain.
Toxicon. 2008 Sep 15;52(4):541-50. doi: 10.1016/j.toxicon.2008.06.028. Epub 2008 Jul 9.
The suitability and sensitivity of two neural cell models, NG108-15 and Neuro-2a, to different marine toxins were evaluated under different incubation and exposure times and in the presence or absence of ouabain and veratridine (O/V). NG108-15 cells were more sensitive to pectenotoxin-2 than Neuro-2a cells. For saxitoxin, brevetoxin-3, palytoxin, okadaic acid and dinophysistoxin-1 both cell types proved to be sensitive and suitable for toxicity evaluation. For domoic acid preliminary results were presented. Setting incubation time and exposure time proved to be critical for the development of the assays. In order to reduce the duration of the assays, it was better to reduce cell time incubation previous to toxin exposure than exposure time. For palytoxin, after 24h of growth, both cell types were sensitive in the absence of O/V. When growth time previous to toxin exposure was reduced, both cell types were unsensitive to palytoxin when O/V was absent. Although dinophysistoxin-1 and okadaic acid are both phosphatase inhibitors, these toxins did not respond similarly in front of the experimental conditions studied. Both cell types were able to identify Na-channel acting toxins and allowed to quantify the effect of saxitoxin, brevetoxin-3, palytoxin, okadaic acid, dinophysistoxin-1 and pectenotoxin-2 under different experimental conditions.
在不同的孵育和暴露时间以及存在或不存在哇巴因和藜芦定(O/V)的情况下,评估了两种神经细胞模型NG108-15和Neuro-2a对不同海洋毒素的适用性和敏感性。NG108-15细胞对pectenotoxin-2比Neuro-2a细胞更敏感。对于石房蛤毒素、短裸甲藻毒素-3、刺尾鱼毒素、冈田酸和鳍藻毒素-1,两种细胞类型都被证明对毒性评估敏感且适用。对于软骨藻酸,给出了初步结果。设定孵育时间和暴露时间被证明对检测的开展至关重要。为了缩短检测时间,减少毒素暴露前的细胞孵育时间比减少暴露时间更好。对于刺尾鱼毒素,在生长24小时后,在不存在O/V的情况下两种细胞类型都敏感。当毒素暴露前的生长时间减少时,在不存在O/V的情况下两种细胞类型对刺尾鱼毒素都不敏感。尽管鳍藻毒素-1和冈田酸都是磷酸酶抑制剂,但在研究的实验条件下,这些毒素的反应并不相似。两种细胞类型都能够识别作用于钠通道的毒素,并能够在不同实验条件下对石房蛤毒素、短裸甲藻毒素-3、刺尾鱼毒素、冈田酸、鳍藻毒素-1和pectenotoxin-2的作用进行定量。