Validation of an Avian Model of Induced-Thyrotoxicosis

T. Hill
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Abstract

TYLER P. HILL, and HOLLY RACINE, Dept of Biomedical Science, West Liberty University, West Liberty, WV, 26074, Validation of an Avian Model of Induced-Thyrotoxicosis This project aimed to validate the avian model developed in our lab with the ultimate goal of discovering the potential link between maternal hyperthyroidism and craniosynostosis (CS). The method of validation for this model was to qualitatively measure the levels of thyroxine (T4) present in the treated embryos' systems at different time points post-injection. T4 was fluorescently labeled using Alexa fluorophore 488 and was injected into an experimental group of N=37 on embryonic day 11 (E11). Three control groups were used one N=9 Saline injected, another N=5 injected with the unconjugated free dye, and lastly, N=5 T4 injected only.    The results showed that after 24 hours post-injection the fluorescent tag was present in the albumen with a peak at 72 hours, after 48 hours fluorescence is detected in the yolk with a peak at 96 hours, finally, after 72 hours there is a large peak in the blood before becoming undetectable at 96 hours. These trends suggest the injection is traveling from the injection site through these tissues and ultimately into the bloodstream, which is supported by the molecular assaying via ELISAs previously conducted. The results of this project in conjunction with the ELISA data show that the peaks detected are from injected T4 rather than a natural spike in development. Prior physiological data collected supports that the T4 injection not only makes its way into the bloodstream but also has a metabolic effect leading to changes in development. These results combined validate this model of induced thyrotoxicosis. (Supported by NIH Grant P20GM103434 to the West Virginia IDeA Network for Biomedical Research Excellence) Acknowledgment of NASA West Virginia Space Grant Consortium (Grant #80NSSC20M0055)
鸡诱导性甲状腺毒症模型的验证
TYLER P. HILL和HOLLY RACINE, West Liberty大学生物医学科学系,West Liberty, WV, 26074,诱导性甲状腺毒症鸟类模型的验证本项目旨在验证我们实验室开发的鸟类模型,最终目的是发现母体甲状腺功能亢进与颅骨闭锁(CS)之间的潜在联系。该模型的验证方法是定性测量注射后不同时间点处理胚胎系统中甲状腺素(T4)的水平。用Alexa荧光团488对T4进行荧光标记,于胚胎第11天(E11)注射到N=37的实验组。3个对照组,N=9注射生理盐水,N=5注射未偶联游离染料,N=5只注射T4。结果表明,注射后24小时,蛋白中出现荧光标记,72小时出现荧光峰,48小时后,蛋黄中出现荧光,96小时出现荧光峰,72小时后,血液中出现一个大峰,96小时时无法检测到荧光。这些趋势表明,注射是从注射部位经过这些组织并最终进入血液,这得到了先前通过elisa进行的分子分析的支持。本项目的结果与ELISA数据相结合表明,检测到的峰值来自注射T4,而不是发育过程中的自然峰值。先前收集的生理学数据支持T4注射不仅进入血液,而且具有导致发育变化的代谢作用。这些结果共同验证了这种诱导甲状腺毒症模型。(由NIH资助P20GM103434向西弗吉尼亚IDeA网络提供卓越生物医学研究支持)
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