Gallagher E P, Gross T S, Sheehy K M
Department of Physiological Sciences, Center for Environmental and Human Toxicology, University of Florida, Gainesville, FL 32611-0885, USA.
Aquat Toxicol. 2001 Nov 12;55(3-4):223-37. doi: 10.1016/s0166-445x(01)00158-8.
A number of freshwater lakes and reclaimed agricultural sites in Central Florida have been the receiving waters for agrochemical and municipal runoff. One of these sites, Lake Apopka, is also a eutrophic system that has been the focus of several case studies reporting altered reproductive activity linked to bioaccumulation of persistent organochlorine chemicals in aquatic species. The present study was initiated to determine if brown bullheads (Ameiurus nebulosus) from the north marsh of Lake Apopka (Lake Apopka Marsh) exhibit an altered capacity to detoxify environmental chemicals through hepatic glutathione S-transferase (GST)-mediated conjugation as compared with bullheads from a nearby reference site (Lake Woodruff). We also compared plasma sex hormone concentrations (testosterone, 17-beta estradiol, and 11 keto-testosterone) in bullheads from the two sites. Female bullheads from Lake Apopka had 40% lower initial rate GST conjugative activity toward 1-chloro-2,4-dinitrobenzene (CDNB), 50% lower activity towards p-nitrobutyl chloride (NBC), 33% lower activity toward ethacrynic acid (ECA), and 43% lower activity toward Delta5-androstene-3,17-dione (Delta(5)-ADI), as compared with female bullheads from Lake Woodruff. Enzyme kinetic analyses demonstrated that female bullheads from Lake Apopka had lower GST-catalyzed CDNB clearance than did female Lake Woodruff bullheads. Western blotting studies of bullhead liver cytosolic proteins demonstrated that the reduced GST catalytic activities in female Lake Apopka bullheads were accompanied by lower expression of hepatic GST protein. No site differences were observed with respect to GST activities or GST protein expression in male bullheads. Female Lake Apopka bullheads also had elevated concentrations of plasma androgens (testosterone and 11-ketotestosterone) as compared with females from Lake Woodruff. In contrast, male Lake Apopka bullheads had elevated levels of plasma estrogen but similar levels of androgens as compared with male bullheads from Lake Woodruff. Collectively, our studies indicate the presence of reduced GST protein expression, reduced GST conjugative capacity and altered sex steroid homeostasis in female bullheads from a contaminated field site in Central Florida. The implications of these physiological alterations in terms of pollutant biotransformation and reproduction are discussed.
佛罗里达州中部的一些淡水湖和开垦的农业用地一直是农用化学品和城市径流的受纳水体。其中一个地点,阿波普卡湖,也是一个富营养化系统,它是几个案例研究的重点,这些研究报告了与水生物种中持久性有机氯化学物质的生物积累相关的生殖活动改变。本研究旨在确定来自阿波普卡湖(阿波普卡湖沼泽)北部沼泽的褐首鲶(Ameiurus nebulosus)与来自附近参考地点(伍德拉夫湖)的鲶鱼相比,是否通过肝脏谷胱甘肽S-转移酶(GST)介导的结合作用表现出改变的环境化学物质解毒能力。我们还比较了两个地点鲶鱼的血浆性激素浓度(睾酮、17-β雌二醇和11-酮睾酮)。与来自伍德拉夫湖的雌性褐首鲶相比,来自阿波普卡湖的雌性褐首鲶对1-氯-2,4-二硝基苯(CDNB)的初始速率GST结合活性低40%,对氯硝基丁酯(NBC)的活性低50%,对依他尼酸(ECA)的活性低33%,对Δ5-雄烯-3,17-二酮(Δ5-ADI)的活性低43%。酶动力学分析表明,来自阿波普卡湖的雌性褐首鲶的GST催化的CDNB清除率低于来自伍德拉夫湖的雌性褐首鲶。对褐首鲶肝脏胞质蛋白的蛋白质印迹研究表明,阿波普卡湖雌性褐首鲶中GST催化活性的降低伴随着肝脏GST蛋白表达的降低。在雄性褐首鲶中未观察到GST活性或GST蛋白表达的地点差异。与来自伍德拉夫湖的雌性相比,阿波普卡湖的雌性褐首鲶血浆雄激素(睾酮和11-酮睾酮)浓度也升高。相比之下,与来自伍德拉夫湖的雄性褐首鲶相比,阿波普卡湖的雄性褐首鲶血浆雌激素水平升高,但雄激素水平相似。总体而言,我们的研究表明,来自佛罗里达州中部受污染现场的雌性褐首鲶存在GST蛋白表达降低、GST结合能力降低和性类固醇稳态改变的情况。讨论了这些生理改变在污染物生物转化和繁殖方面的意义。