Development of a serum vitellogenin MRM biomarker for determining sex in American eel (Anguilla rostrata)

鳗鱼的性别决定并不像人类那样固定，雄性与雌性的比例也是未知的。 比如环境因素。在商业密度下生长的幼鳗（玻璃/幼鳗）显示， 主要是男性（95%）。对于商业鳗鱼养殖，雌鳗将达到预期的市场规模 （>300 g），而雄性成熟时尺寸较小（<200 g），无法实现有利可图的市场规模。成功 鳗鱼养殖业的未来发展，将取决于操纵鳗鱼的性别比例，以增加雌性的比例。一些 研究人员已经证明，使用β-雌二醇和其他物质对欧洲和日本鳗鱼进行了性别操纵 雌激素化合物开发操纵鳗鱼性别的方法的困难在于， 改变幼鳗的性别（从1 - 30 g），但直到它们开始 性腺发育成熟（80-150 g）。本项目旨在开发一种具有成本效益的液相色谱- 多反应监测质谱法（LC-MRM-MS/MS）用于开发特定的雌性动物 血液生物标志物，卵黄蛋白原，以确定性别在美国鳗鱼（Anguilla rostrata）在早期阶段比 可通过标准显微镜方法观察到。将对血液进行LC-MRM-MS/MS方法， 在NovaEel的雌性化试验中采集的鳗鱼性腺样本。卵黄原蛋白是一种在体内合成的蛋白质 卵生动物，在肝脏中产生，并通过血液运送到发育中的卵母细胞， 卵巢它已被用于其他鱼类作为雌性的标记。将对数据进行关联和验证 通过显微镜方法确定性腺发育。制定具有成本效益的方法， 使鳗鱼雌性化并快速确定鳗鱼的性别将有助于该公司开发商业上可行的 加拿大东部的鳗鱼养殖业

Sex determination in eels is not fixed as in humans and the ratio of males to females is subject to unknown factors, such as environmental cues. Juvenile (glass/elvers) eels grown at commercial densities, were shown to predominantly end up as males (95%). For commercial eel farming, female eels will reach desired market size (>300 g), whereas males mature at a smaller size (<200 g) and do not achieve profitable market size. Success of eel farming will depend on manipulating the sex ratio of eels to increase the proportion of females. Some researchers have demonstrated sex manipulation in European and Japanese eels using ß-estradiol and other estrogenic compounds. The difficulty in developing methods to manipulate sex in eels is that it is best to change the sex in young eels (from 1 -30 g), but it is difficult to determine the sex of eels until they begin to develop maturing gonads (80-150 g). This project intends to develop a cost-effective Liquid Chromatography - Multiple Reaction Monitoring Mass Spectrometry (LC-MRM-MS/MS) method to develop a specific female blood biomarker, vitellogenin, to determine sex in the American eel (Anguilla rostrata) at an earlier stage than is observable by standard microscopic methods. LC-MRM-MS/MS methods will be carried out on blood, and gonad samples from eels sampled in NovaEel's feminization trials. Vitellogenin is a protein synthesized in oviparous animals that is made in the liver and carried in the blood to be delivered to the developing oocytes in the ovaries. It has been used in other fish species as a marker for females. Data will be correlated and validated with gonad development as determined by microscopic methods. Development of cost-effective methods to feminize eels and to rapidly determine the sex of eels will help the company to develop commercially viable eel farming in Eastern Canada.