Moncaut Natalia, Nostro Fabiana Lo, Maggese María Cristina
Laboratorio de Embriología Animal, Departamento de Biodiversidad y Biología Experimental, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, Pab. II, 4to piso C1428EHA, Buenos Aires, Argentina.
Aquat Toxicol. 2003 Apr 10;63(2):127-37. doi: 10.1016/s0166-445x(02)00175-3.
During the last decade, special attention has been focused on the consequences of exposure to environmental estrogens on reproduction in wild fish species. For this reason, characterization of biomarkers of such exposures could result in useful tools for these studies. The detection of vitellogenin (Vtg), a precursor of yolk proteins, is being intensely studied since its synthesis in the liver is regulated by the estradiol-17beta and is influenced by other estrogenic compounds. The aim of this work was to assess the presence of Vtg in the surface mucus of males of Cichlasoma dimerus (Teleostei, Perciformes), a typical South American freshwater cichlid, after hormonal treatment with estradiol-17beta (intraperitoneal injections of 10 microg E(2)/g fish). Plasma and mucus from females and treated males analyzed by Western blot revealed that different heterologous antisera against Vtg bind to putative protein of 180 & 120 kDa and 120 & 110 kDa, respectively, whereas no reaction was found in samples of untreated males. The same profile was observed in mucus samples using Dot blot, a very easy and direct technique. Using immunocytochemistry techniques, immunoreactive Vtg (ir-Vtg) producing cells in the liver of females and treated males were detected. Testes and liver tissues were also assessed by histological techniques. Marked alterations in both organs were observed, such as lower sperm production, presence of immature germ cells in the lobular lumen of testes, and some morphology changes in the hepatocytes due to the accumulation of Vtg. This is the first report about the effects of an estrogen in the Vtg synthesis and their consequences on liver and gonads of a South American fresh water cichlid. These results also support the possibility of using Vtg from surface mucus as a potential biomarker for estrogenic compounds through a noninvasive, useful and easy assay to monitor the presence of endocrine disrupting chemicals in the environment.
在过去十年中,人们特别关注环境雌激素暴露对野生鱼类繁殖的影响。因此,表征此类暴露的生物标志物可能会为这些研究带来有用的工具。卵黄蛋白原(Vtg)是卵黄蛋白的前体,由于其在肝脏中的合成受17β-雌二醇调节且受其他雌激素化合物影响,因此对其检测正在进行深入研究。本研究的目的是评估在南美淡水慈鲷——双斑丽体鱼(硬骨鱼纲,鲈形目)雄性个体经17β-雌二醇激素处理(腹腔注射10μg E₂/g鱼)后,其体表黏液中是否存在Vtg。通过蛋白质免疫印迹法分析雌性和经处理雄性的血浆及黏液发现,针对Vtg的不同异源抗血清分别与推定的180 kDa和120 kDa以及120 kDa和110 kDa的蛋白质结合,而未处理雄性的样本中未发现反应。使用斑点印迹法(一种非常简便直接的技术)在黏液样本中也观察到了相同的图谱。采用免疫细胞化学技术,在雌性和经处理雄性的肝脏中检测到了产生免疫反应性Vtg(ir-Vtg)的细胞。还通过组织学技术评估了睾丸和肝脏组织。观察到两个器官都有明显变化,如精子产量降低、睾丸小叶腔中存在未成熟生殖细胞以及由于Vtg积累导致肝细胞出现一些形态变化。这是关于雌激素对南美淡水慈鲷Vtg合成的影响及其对肝脏和性腺后果的首次报道。这些结果还支持通过一种无创、有用且简便的检测方法,将体表黏液中的Vtg用作雌激素化合物潜在生物标志物的可能性,以监测环境中内分泌干扰化学物质的存在。