NGF、NT3和BDNF在鹅等级前卵泡中的表达规律研究

发布时间：2018-11-27 06:46

【摘要】：鹅多数品种具有明显的季节性繁殖与及强烈的就巢性等特点,严重制约了鹅业发展。籽鹅是世界高产蛋鹅品种之一,因其体型小、产蛋多等特点,素有“产蛋王”之美称。家禽属于卵生动物,在适宜环境条件下,卵巢上的卵泡可持续发育、成熟、排卵,发育到性成熟的时候受精,且家禽排卵后不会有黄体形成,是较为理想的动物试验模型。本试验以籽鹅作为试验动物,围绕鹅等级前卵泡发育调控机制展开研究。神经营养因子家族((Neurotrophic factors,NTFs)属于蛋白质类大分子,由多种神经营养因子组成,其中脑源性神经营养因子(Brain-derived neurotrophie factor,BDNF))、神经生长因子(Nerve growth factor.NGF),神经营养因子-3(Neurotrophin-3)是该家族中的重要成员,同时也是发现最早、当前研究较为热门的神经营养因子。现有研究表明,NGF、NT-3以及BDNF与其受体TrkB基因不仅对神经细胞具有调控作用,在心血管、内分泌、免疫以及繁殖等系统中也有相应的表达。在鹅卵泡发育过程中神经营养因子有关基因及其代谢产物起着重要作用,为了解神经营养因子以及具有调控作用基因的生物学信息以及相关基因的表达趋势,本研究采用高通量转录组测序技术。它是组织或细胞不同生理状态下转录出来的所有RNA的总和。本试验主要以籽鹅等级前卵泡作为研究对象,选用新一代Illumina Hiseq 2500高通量测序平台对提取的籽鹅等级前卵泡(5个等级)的总RNA进行转录组测序。本次转录组测序,共得到长度在150 bp左右的reads上百万条。鹅归属非模式生物,尚未有参考基因组,通过对鹅等级前卵泡优质序列的拼接组装以及GO、COG数据库注释,挖掘出与繁殖、代谢、生长速度甚至抗病等相关的候选基因,为以后实践分子育种以及功能基因研究提供理论基础;KEGG pathway注释到ERK信号通路以及该通路上的主要基因,计算FPKM值作为基因的表达量,利用RT-PCR和Western Blot技术,分析鹅等级前卵泡中具有主要调控作用的NT3、NGF、BDNF及其受体TrkB基因的表达,结果如下:4个基因在鹅5个等级前卵泡中均有表达,其中在小白卵泡中的表达量最多。
[Abstract]:Most goose varieties have obvious characteristics of seasonal reproduction and strong nesting, which seriously restrict the development of goose industry. Seed goose is one of the high-yielding egg geese in the world. Poultry belongs to oviparous animal. Under suitable environment, follicle on ovary can develop continuously, mature, ovate, fertilize when developing to sexual maturity, and there will not be luteal body formation after ovulation of poultry. It is an ideal animal experimental model. In this study, the control mechanism of prefollicular development of geese was studied with Seedling Goose as experimental animal. The family of neurotrophic factors (Neurotrophic factors,NTFs) belongs to protein macromolecules and consists of a variety of neurotrophic factors, including Brain-derived neurotrophie factor,BDNF), neurotrophic factor (Brain-derived neurotrophie factor,BDNF), neurotrophic factor (Nerve growth factor.NGF), and neurotrophic factor (Nerve growth factor.NGF). Neurotrophic factor-3 (Neurotrophin-3) is an important member of the family and is the most popular neurotrophic factor. Current studies have shown that NGF,NT-3, BDNF and its receptor TrkB genes not only regulate nerve cells, but also express them in cardiovascular, endocrine, immune and reproductive systems. Neurotrophin-related genes and their metabolites play an important role in the development of goose follicles. In order to understand the biological information of neurotrophic factors and regulatory genes and the expression trend of related genes, neurotrophic factors and their related genes play an important role in the development of goose follicles. High-throughput transcriptome sequencing was used in this study. It is the sum of all RNA transcribed by tissues or cells in different physiological states. A new generation of Illumina Hiseq 2500 high throughput sequencing platform was used to sequence the total RNA of prefollicles (5 grades) extracted from the seed geese. The transcriptome was sequenced and millions of reads about 150 bp in length were obtained. Geese belong to non-model organisms without reference genomes. Candidate genes related to reproduction, metabolism, growth rate and even disease resistance are mined by splicing and assembling high-quality pre-follicular sequences and annotating GO,COG database. It provides a theoretical basis for the practice of molecular breeding and functional gene research in the future. KEGG pathway annotates the ERK signaling pathway and the major genes in the pathway, calculates the FPKM value as the expression amount of the gene, and uses RT-PCR and Western Blot techniques to analyze the NT3,NGF, which plays a major role in the pre-grade follicles of goose. The expression of BDNF and its receptor TrkB gene was as follows: the four genes were expressed in goose prefollicles of 5 grades, and most of them were expressed in small white follicles.
【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S835

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