【摘要】：鵝多數(shù)品種具有明顯的季節(jié)性繁殖與及強烈的就巢性等特點,嚴重制約了鵝業(yè)發(fā)展。籽鵝是世界高產(chǎn)蛋鵝品種之一,因其體型小、產(chǎn)蛋多等特點,素有“產(chǎn)蛋王”之美稱。家禽屬于卵生動物,在適宜環(huán)境條件下,卵巢上的卵泡可持續(xù)發(fā)育、成熟、排卵,發(fā)育到性成熟的時候受精,且家禽排卵后不會有黃體形成,是較為理想的動物試驗模型。本試驗以籽鵝作為試驗動物,圍繞鵝等級前卵泡發(fā)育調控機制展開研究。神經(jīng)營養(yǎng)因子家族((Neurotrophic factors,NTFs)屬于蛋白質類大分子,由多種神經(jīng)營養(yǎng)因子組成,其中腦源性神經(jīng)營養(yǎng)因子(Brain-derived neurotrophie factor,BDNF))、神經(jīng)生長因子(Nerve growth factor.NGF),神經(jīng)營養(yǎng)因子-3(Neurotrophin-3)是該家族中的重要成員,同時也是發(fā)現(xiàn)最早、當前研究較為熱門的神經(jīng)營養(yǎng)因子�，F(xiàn)有研究表明,NGF、NT-3以及BDNF與其受體TrkB基因不僅對神經(jīng)細胞具有調控作用,在心血管、內分泌、免疫以及繁殖等系統(tǒng)中也有相應的表達。在鵝卵泡發(fā)育過程中神經(jīng)營養(yǎng)因子有關基因及其代謝產(chǎn)物起著重要作用,為了解神經(jīng)營養(yǎng)因子以及具有調控作用基因的生物學信息以及相關基因的表達趨勢,本研究采用高通量轉錄組測序技術。它是組織或細胞不同生理狀態(tài)下轉錄出來的所有RNA的總和。本試驗主要以籽鵝等級前卵泡作為研究對象,選用新一代Illumina Hiseq 2500高通量測序平臺對提取的籽鵝等級前卵泡(5個等級)的總RNA進行轉錄組測序。本次轉錄組測序,共得到長度在150 bp左右的reads上百萬條。鵝歸屬非模式生物,尚未有參考基因組,通過對鵝等級前卵泡優(yōu)質序列的拼接組裝以及GO、COG數(shù)據(jù)庫注釋,挖掘出與繁殖、代謝、生長速度甚至抗病等相關的候選基因,為以后實踐分子育種以及功能基因研究提供理論基礎;KEGG pathway注釋到ERK信號通路以及該通路上的主要基因,計算FPKM值作為基因的表達量,利用RT-PCR和Western Blot技術,分析鵝等級前卵泡中具有主要調控作用的NT3、NGF、BDNF及其受體TrkB基因的表達,結果如下:4個基因在鵝5個等級前卵泡中均有表達,其中在小白卵泡中的表達量最多。
[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.
【學位授予單位】：吉林農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S835

【參考文獻】

相關期刊論文 前10條

1 馮月枝;邱綺;楊冬梓;張清學;;新生大鼠卵巢體外培養(yǎng)中NT3與LIF對始基卵泡發(fā)育的影響[J];中山大學學報(醫(yī)學科學版);2016年04期

2 趙建華;唐金海;;乳腺癌基礎研究與精準醫(yī)學[J];中國腫瘤外科雜志;2015年03期

3 胡民強;郭金彪;張建華;;鵝反季節(jié)繁殖生產(chǎn)技術[J];安徽農(nóng)業(yè)科學;2013年20期

4 金艷梅;;顆粒細胞對卵泡發(fā)育的影響[J];中國畜牧獸醫(yī);2010年08期

5 林鵬飛;郝元斌;郭慧利;劉紅林;芮榮;;豬分離卵泡體外培養(yǎng)過程中Fas/FasL對顆粒細胞凋亡的作用[J];動物學研究;2010年03期

6 陳士林;何柳;劉明珠;錢俊;閆海霞;程翔林;錢忠直;;本草基因組方法學研究[J];世界科學技術(中醫(yī)藥現(xiàn)代化);2010年03期

7 馬永和;孫永峰;李純錦;周虛;;神經(jīng)生長因子受體TrkA在母牛性腺及性腺外生殖器官中的表達研究[J];吉林農(nóng)業(yè)大學學報;2010年03期

8 王曉麗;楊娟;陳玉;成俊萍;潘紅平;韋精衛(wèi);陸鳳花;石德順;;水牛卵丘顆粒細胞凋亡與卵母細胞體外發(fā)育潛能的研究[J];畜牧獸醫(yī)學報;2009年09期

9 陳秀萍;陳鋒劍;姜勛平;丁家桐;;鵝卵泡顆粒細胞凋亡與卵黃E_2和P_4水平關系的研究[J];揚州大學學報(農(nóng)業(yè)與生命科學版);2008年02期

10 施振旦;黃運茂;吳偉;;鵝產(chǎn)蛋周期及其生理學調控機制研究的回顧[J];中國家禽;2008年09期

相關博士學位論文 前6條

1 謝偉林;強直性脊柱炎發(fā)病的免疫遺傳機制及風險預報研究[D];第二軍醫(yī)大學;2012年

2 孫永峰;神經(jīng)營養(yǎng)因子4及其受體在母牛生殖系統(tǒng)中的表達與部分功能研究[D];吉林大學;2011年

3 林金杏;局部性促生長因子對雞卵泡發(fā)育的調控及其機理的研究[D];浙江大學;2011年

4 馬永和;神經(jīng)生長因子及其受體在母牛生殖系統(tǒng)中的表達與功能研究[D];吉林大學;2010年

5 易康樂;Figla和BDNF對豬和牛卵母細胞及早期胚胎生長發(fā)育的影響[D];吉林大學;2008年

6 金艷梅;雞等級前卵泡顆粒細胞發(fā)育的激素和營養(yǎng)調控及機理研究[D];浙江大學;2007年

相關碩士學位論文 前8條

1 王超;萎縮芽孢桿菌(Bacillus atrophaeus)與鈾、鉻相互作用機理研究[D];西南科技大學;2015年

2 侯曉峰;神經(jīng)生長因子對小鼠卵巢顆粒細胞增殖及孕酮分泌的影響[D];吉林大學;2014年

3 朱海洋;NGF和BDNF mRNA在鵝等級前卵泡中表達規(guī)律的研究[D];吉林農(nóng)業(yè)大學;2014年

4 瞿長偉;神經(jīng)生長因子及其受體在蛋雞等級卵泡和輸卵管上的表達研究[D];南京農(nóng)業(yè)大學;2012年

5 朱曉玲;神經(jīng)營養(yǎng)因子NGF、BDNF、NT-4/5在雌性水牛生殖器官的表達研究[D];吉林大學;2011年

6 李r，

本文編號：2359738