Chandler G Thomas, Volz David C
Department of Environmental Health Sciences, Norman J. Arnold School of Public Health, University of South Carolina, Columbia, SC 29208, USA.
Mar Biotechnol (NY). 2004 Mar-Apr;6(2):128-37. doi: 10.1007/s10126-002-0105-y. Epub 2004 Apr 19.
Confocal laser scanning microscopy (CLSM) represents a powerful, but largely unexplored ecotoxicologic tool for rapidly assessing in vivo effects of toxicants on marine invertebrate embryo quality and development. We describe here a new semiquantitative CLSM approach for assessing relative yolk quantity in marine invertebrate embryos (harpacticoid copepods) produced by parents reared from hatching to adult in the polycylic aromatic hydrocarbon chrysene. This method is based on fluorogenic labeling of embryo yolk and subsequent statistical analysis of areal pixel intensities over multiple Z-series using a general linear model (GLM)-nested analysis of variance. The fluorescent yolk-labeling method described here was able to detect statistically significant differences in yolk concentrations in marine copepod (Amphiascus tenuiremis) eggs or embryos from females exposed to ultraviolet light and chrysene-contaminated sediments. Yolk intensities in embryos from females cultured throughout their life cycles in clean sediments were statistically identical with or without UV exposure. In contrast, yolk intensities in embryos of females cultured throughout their life cycle in chrysene-contaminated sediments were significantly higher in the non-UV-exposed treatment with chrysene at 2500 ng/g sediment (65.7% higher) and the UV-exposed treatment with chrysene at 500 ng/g sediment (76.6% higher).
共聚焦激光扫描显微镜（CLSM）是一种强大但在很大程度上未被探索的生态毒理学工具，可用于快速评估有毒物质对海洋无脊椎动物胚胎质量和发育的体内效应。我们在此描述一种新的半定量CLSM方法，用于评估由在多环芳烃chrysene中从孵化饲养至成年的亲体产生的海洋无脊椎动物胚胎（猛水蚤桡足类）中的相对卵黄量。该方法基于胚胎卵黄的荧光标记以及随后使用通用线性模型（GLM）-嵌套方差分析对多个Z系列上的区域像素强度进行统计分析。本文所述的荧光卵黄标记方法能够检测出暴露于紫外线和chrysene污染沉积物的雌性海洋桡足类（Amphiascus tenuiremis）卵或胚胎中卵黄浓度的统计学显著差异。在清洁沉积物中终生培养的雌性所产胚胎中的卵黄强度，无论有无紫外线暴露，在统计学上都是相同的。相比之下，在chrysene污染沉积物中终生培养的雌性所产胚胎中的卵黄强度，在沉积物中chrysene含量为2500 ng/g的非紫外线暴露处理中显著更高（高65.7%），在沉积物中chrysene含量为500 ng/g的紫外线暴露处理中也显著更高（高76.6%）。
