摘要
目的以鲫鱼Carassius auratus胚胎为动物模型,观察孕酮对其形态发育、昼夜节律和下丘脑-垂体-性腺(HPG)轴相关基因表达的影响。方法将性成熟的鲫鱼通过干法人工授精获取受精卵,将胚胎暴露于环境相关浓度的孕酮(P4)中,采用倒置显微镜观察胚胎的形态,并统计胚胎的存活率、畸形率和孵出率,通过实时荧光定量PCR检测HPG轴和昼夜节律信号相关基因的表达。结果与对照组相比,各浓度P4暴露组胚胎的存活率、孵出率、畸形率并无显著差异。高浓度P4暴露显著诱导了昼夜节律相关基因Clock1、Cry1、Arntl1的表达。P4暴露组上调了HPG轴相关基因Gnrh3、Lhβ的表达,并进一步调节类固醇生成相关基因Cyp11a、Cyp19a、Hsd3β、Hsd20β的表达水平,导致了昼夜节律和内分泌系统的紊乱。结论P4对鲫鱼胚胎发育没有明显的致死、致畸效应,环境相关浓度的P4干扰了其HPG轴和昼夜节律相关基因的表达,从而可能导致细胞和生理代谢的紊乱,进而影响鲫鱼幼鱼的性腺分化和性别比例。
Objective To explore the effects of progesterone on the morphological development and the expression of genes related to circadian rhythm signaling and the hypothalamic-pituitary gonadal(HPG)axis of Carassius auratus embryos.Methods The fertilized eggs of C.auratus were obtained by dry artificial insemination.The embryos were exposed to progesterone(P4)at environmentally relevant concentrations,and then the embryonic morphology was observed by inverted microscope,followed by calculating the survival rate,malformation rate and hatching rate.Finally,qRT-PCR was used to detect the expression of genes related to circadian rhythm signaling and HPG axis.Results Compared with the control group,there were no significant differences in the survival rate,hatching rate and malformation rate of P4-exposed groups.The highest concentration of P4 exposure led to a significant up-regulation of circadian rhythm related genes Clock1,Cry1 and Arntl1.In addition,P4 treatment increased the expression levels of HPG axis-related genes Gnrh3 and Lhβ,and further regulated the expressions of steroidogenesis related genes Cyp11a,Cyp19a,Hsd3β,and Hsd20β,leading to the disturbance of circadian rhythm and endocrine system.Conclusion P4 at environmentally relevant concentrations has no obvious effects on the death and teratogenesis of C.auratus embryos,but can interfere with the expression of HPG axis and circadian rhythm-related genes.This may lead to the disorder of cell and physiological metabolism,and then influence the gonadal differentiation and sex ratio of juvenile fish.
作者
王晨
满晓婷
王颖
张云紫
周新雨
万汝燕
夏晓华
WANG Chen;MAN Xiaoting;WANG Ying;ZHANG Yunzi;ZHOU Xinyu;WAN Ruyan;XIA Xiaohua(College of Life Science,Henan Normal University,Xinxiang,Henan Province 453007,China)
出处
《四川动物》
北大核心
2023年第1期46-53,共8页
Sichuan Journal of Zoology
基金
国家自然科学基金项目(31972773)
河南省自然科学基金项目(222300420471)
河南省重点研发与推广专项(科技攻关)(212102110379)
河南师范大学面上培育基金项目(2021PL16)。
关键词
鲫鱼
孕酮
胚胎
昼夜节律
HPG轴
Carassius auratus
progesterone
embryos
circadian rhythm
HPG axis
