摘要
【目的】明确FoxO3转录因子在不同日龄鸡卵巢组织中的表达模式及其具体定位情况,为后续开展家禽卵泡激活及发育调控等相关机理研究提供科学依据。【方法】在GenBank中搜索鸡与其他物种(鸭、鹅、老鼠、猪、牛及人类)的FoxO3氨基酸序列,通过LaserGene分析鸡FoxO3氨基酸序列与其他物种FoxO3氨基酸序列间的亲缘关系及同源差异情况;利用RT-PCR鉴定FoxO3基因是否在鸡卵巢组织中表达,再采用实时荧光定量PCR检测不同发育阶段鸡卵巢组织中FoxO3基因的表达水平,最后以免疫荧光检测FoxO3蛋白在鸡卵巢组织中的定位情况。【结果】鸡与鸭和鹅的FoxO3氨基酸序列相似性分别为92.7%和95.3%,基于FoxO3氨基酸序列相似性构建的系统发育进化树也显示鸡与鸭和鹅的亲缘关系较近,说明FoxO3基因的进化相对较保守。在不同发育阶段的鸡卵巢组织中均能检测到FoxO3基因表达,且FoxO3基因在0日龄鸡卵巢组织中的相对表达量最高,显著高于在其他发育阶段的相对表达量(P<0.05)。在0日龄鸡卵巢组织中未检测到FoxO3蛋白,但在21日龄和成年鸡的卵巢组织中均能检测到FoxO3蛋白;在21日龄鸡卵巢组织中FoxO3蛋白主要定位在原始卵泡细胞周围,在卵泡内部没有表达;而在成年鸡卵巢组织中FoxO3蛋白仅定位于大卵泡细胞边缘,小卵泡细胞内并未发现FoxO3蛋白。【结论】由于鸡和哺乳动物的FoxO3氨基酸序列高度同源,且FoxO3蛋白在鸡卵巢组织中的定位与在哺乳动物卵巢组织中的定位相似,说明FoxO3在鸡卵巢组织中发挥着与哺乳动物相似的功能,即在卵泡激活与成熟过程中发挥重要作用,因此通过调控FoxO3能有效提高鸡的繁殖性能。
【Objective】The expression pattern and specific localization of FoxO3 transcription factor in ovarian tissue of chickens at different ages were determined,providing a basis for the subsequent studies on related mechanisms such as follicular activation and developmental regulation in poultry.【Method】FoxO3 amino acid sequences of chicken and other species(duck,goose,mouse,pig,ox and human)were searched in GenBank,and the genetic relationship and homologous difference between FoxO3 amino acid sequence of chicken and FoxO3 amino acid sequence of other species were analyzed by Laser Gene.RT-PCR was used to identify whether FoxO3 gene was expressed in chicken ovarian tissue.Then realtime fluorescence quantitative PCR was used to detect FoxO3 gene expression level in chicken ovarian tissue at different stages.Finally,immunofluorescence was used to detect FoxO3 protein localization in chicken ovarian tissue.【Result】The similarity of FoxO3 amino acid sequence between chicken and duck and goose was 92.7%and 95.3%,respectively.The phylogenetic tree based on T-PCR similarity also showed that chicken was closely related to duck and goose,indicating that FoxO3 gene was relatively conservative in evolution.FoxO3 gene expression could be detected in different stages of chicken ovarian tissue,and the relative expression level of FoxO3 gene in 0-day-old chicken ovarian tissue was the highest,significantly higher than the relative expression level in other stages(P<0.05).FoxO3 protein was not detected in the ovarian tissue of 0-day-old chickens,but FoxO3 protein was detected in the ovarian tissue of 21-day-old and adult chickens.FoxO3 protein was mainly localized around the original follicle cells in the ovarian tissue of chicken at 21 days old,but was not expressed in the follicle.FoxO3 was found only at the edges of large follicular cells in adult chicken ovarian tissue,but not in small follicular cells.【Conclusion】Since the chicken and mammals FoxO3 amino acid sequence has high homology,the location in the chicken and FoxO3 protein in ovarian tissue is similar to the location in the ovarian tissue in mammals,which indicates thatFoxO3 in chicken ovary organization plays a similar function as mammals.The follicle plays an important role in the process of activation and mature,so through regulating FoxO3 can effectively increase the reproductive performance of chicken.
作者
洪永兴
彭腊如
黄振文
徐天鹏
濮黎萍
虞霖田
谢龙
廖玉英
陆阳清
HONG Yong-xing;PENG La-ru;HUANG Zhen-wen;XU Tian-peng;PU Li-ping;YU Lin-tian;XIE Long;LIAO Yu-ying;LU Yang-qing(College of Animal Science and Technology,Guangxi University/State Key Laboratory of Subtropical Agricultural Biological Resources Conservation and Utilization,Nanning 530004,China;Guangxi Agricultural Vocational and Technical College,Nanning 530007,China;Guangxi Animal Husbandry Research Institute,Nanning 530021,China)
出处
《南方农业学报》
CAS
CSCD
北大核心
2020年第8期1849-1856,共8页
Journal of Southern Agriculture
基金
国家自然科学基金项目(31960157)。
关键词
鸡
FoxO3转录因子
卵巢
卵泡激活
表达定位
chicken
FoxO3 transcription factor
ovary
follicle activation
localization of gene expression
