Cell proliferation alterations in Chlorella cells under stress conditions
Cell proliferation alterations in Chlorella cells under stress conditions
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DOI：
10.1016/j.aquatox.2009.07.009
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发表时间：
2009-09-14
影响因子：
4.5
通讯作者：
Cid, Angeles
中科院分区：
文献类型：
--
作者：
Rioboo, Carmen;O'Connor, Jose Enrique;Cid, Angeles
关于藻类的生长和增殖与细胞周期调节的关系知之甚少。当提到污染物对微藻细胞分裂的潜在毒性作用时，知识的缺乏就更严重了。为了评估三嗪除草剂特丁炔对淡水微藻小球藻增殖的影响，使用了三种流式细胞术方法：(1) 使用 5,6-羧基荧光素二乙酸琥珀酰亚胺酯 (CFSE) 染色测量体内细胞分裂，(2) 通过细胞计数确定生长动力学，(3) 使用膜不可渗透的细胞活力评估细胞活力双链核酸染色碘化丙啶(PI)。使用CFSE鉴定微藻细胞后代的生长动力学研究所得结果与细胞计数法测定的结果一致。在所有 C. vulgaris 培养物中，每个母细胞在 96 小时的测定中仅经历一轮分裂，并且细胞分裂发生在黑暗时期。暴露于除草剂的培养物的细胞分裂是异步的。特布丁改变了从每个母细胞获得的子细胞（4 个自孢子）的正常数量。在用 250 nM 处理的培养物中，这一数字仅为 2。接触特丁烯后滞后期的持续时间可能取决于激活细胞质分裂的临界细胞大小的存在。微藻细胞的细胞大小、叶绿素 a 的复杂性和荧光呈现出明显的光/暗（白天/黑夜）循环，但在非分裂 500 nM 培养物中除外，其中特布丁林在循环开始时阻止细胞分裂。活力结果表明，在此浓度下，特丁烯对普通藻细胞具有抑藻作用。通过 CFSE 染色快速、精确地测定细胞增殖，使我们能够开发出一种模型，用于评估普通藻的细胞周期以及污染物在微藻细胞水平上对生长和繁殖的体内影响。 (C) 2009 Elsevier B.V. 保留所有权利。
Very little is known about growth and proliferation in relation to the cell cycle regulation of algae. The lack of knowledge is even greater when referring to the potential toxic effects of pollutants on microalgal cell division. To assess the effect of terbutryn, a triazine herbicide, on the proliferation of the freshwater microalga Chlorella vulgaris three flow cytometric approaches were used: (1) in vivo cell division using 5,6-carboxyfluorescein diacetate succinimidyl ester (CFSE) staining was measured, (2) the growth kinetics were determined by cytometric cell counting and (3) cell viability was evaluated with the membrane-impermeable double-stranded nucleic acid stain propidium iodide (PI).The results obtained in the growth kinetics study using CFSE to identify the microalgal cell progeny were consistent with those determined by cytometric cell counting. In all C. vulgaris cultures, each mother cell had undergone only one round of division through the 96 h of assay and the cell division occurred during the dark period. Cell division of the cultures exposed to the herbicide was asynchronous. Terbutryn altered the normal number of daughter cells (4 autospores) obtained from each mother cell. The number was only two in the cultures treated with 250 nM. The duration of the lag phase after the exposure to terbutryn could be dependent on the existence of a critical cell size to activate cytoplasmic division. Cell size, complexity and fluorescence of chlorophyll a of the microalgal cells presented a marked light/dark (day/night) cycle, except in the non-dividing 500 nM cultures, where terbutryn arrested cell division at the beginning of the cycle. Viability results showed that terbutryn has an algastatic effect in C. vulgaris cells at this concentration.The rapid and precise determination of cell proliferation by CFSE staining has allowed us to develop a model for assessing both the cell cycle of C. vulgaris and the in vivo effects of pollutants on growth and reproduction at microalgal cell level. (C) 2009 Elsevier B.V. All rights reserved.