LUMINOL-DEPENDENT CHEMILUMINESCENCE BY HEMOCYTES OF 2 MARINE BIVALVES, OSTREA-EDULIS AND CRASSOSTREA-GIGAS
LUMINOL-DEPENDENT CHEMILUMINESCENCE BY HEMOCYTES OF 2 MARINE BIVALVES, OSTREA-EDULIS AND CRASSOSTREA-GIGAS
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DOI：
10.3354/dao011173
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发表时间：
1991-10-17
影响因子：
1.4
通讯作者：
MIALHE, E
中科院分区：
文献类型：
--
作者：
BACHERE, E;HERVIO, D;MIALHE, E
使用鲁米诺增强化学发光 (CL) 技术研究了 2 种牡蛎（美味牡蛎和巨牡蛎）血细胞的杀微生物吞噬功能和氧化代谢。首先，在 C. gigas 的血淋巴池上建立了适合海洋双壳类血细胞的实验参数，以获得定性和定量均质的样品，从而进行统计上可比较的测定。使用改良的 Alsever 溶液 (MAS) 可以使血细胞保持非聚集和非刺激状态，直到开始测定。 MAS 终浓度为 3.5% 的大量 2 X 10(5) 血细胞等分试样在酵母聚糖颗粒刺激后显示出高化学发光反应。 80:1 的颗粒:血细胞比例可提供最佳的 CL 活性。活性受到细胞松弛素 B（一种吞噬细胞抑制剂）和叠氮化钠的抑制，后者表明氧产物的参与。确定了合适的参数值后，将 CL 方案应用于 0. edulis，对物种和个体水平的呼吸爆发能力进行定性和定量分析。结果表明 CL 反应的种间和种内差异。在相同的实验条件下，美味O. edulis 血细胞普遍表现出比C gigas 更高的CL 活性。此外，还证明了明显的个体差异。相同实验数量的血细胞在个体之间显示出 CL 反应的巨大差异，这表明可能与血象特征（例如细胞类型百分比）相关。
Microbicidal phagocytic function and oxidative metabolism of the hemocytes of 2 oyster species, Ostrea edulis and Crassostrea gigas, were investigated using a luminol-enhanced chemiluminescence (CL) technique. First, experimental parameters adapted to marine bivalve hemocytes were established on hemolymph pools of C. gigas in order to obtain qualitatively and quantitatively homogeneous samples and so to perform statistically comparable assays. The use of Modified Alsever Solution (MAS) allowed hemocytes to be kept non-aggregated and in a non-stimulated state until starting the assays. A number of aliquots of 2 X 10(5) hemocytes with an MAS final concentration of 3.5% showed high chemiluminescent response after stimulation by zymosan particles. A particle:hemocyte ratio of 80:1 gave optimal CL activity. Activity was inhibited by cytochalasin B, a phagocytic inhibitor, and by sodium azide, the latter indicating the involvement of oxygen products. Having thus defined suitable parameter values, the CL protocol was then applied to 0. edulis for qualitative and quantitative analyses of respiratory burst capacity at the species and individual levels. Results suggested inter- and intra-specific variability of CL responses. Under the same experimental conditions, O. edulis hemocytes generally displayed higher CL activities than C gigas. Moreover, clear individual variability was demonstrated. The same experimental number of hemocytes showed great differences in CL responses between individuals suggesting a possible correlation with hemogram characteristics such as cell type percentages.