本文選題：海蘭褐蛋雞 + FOXL2基因�。� 參考：《河南科技學(xué)院》2017年碩士論文

【摘要】：FOXL2基因是在脊椎動(dòng)物中最先發(fā)現(xiàn)的與卵巢分化有關(guān)的的性別二態(tài)性標(biāo)記,同時(shí)也是人類常染色體基因中第一個(gè)被認(rèn)定在促進(jìn)人的眼瞼的早期發(fā)育、卵巢發(fā)育和維持卵巢功能方面具有重要作用的因子。隨著人們對(duì)于相關(guān)研究的不斷加深,發(fā)現(xiàn)在人的眼瞼、胚胎及成體卵巢顆粒細(xì)胞中FOXL2基因均呈現(xiàn)出特異性的表達(dá),與人的卵巢早衰、卵巢腫瘤、不孕不育、性別決定以及小眼瞼裂綜合征等病癥有關(guān)。FOXL2基因在抑制卵泡凋亡和促進(jìn)卵巢儲(chǔ)備功能方面發(fā)揮著重要作用,其正常表達(dá)對(duì)雞產(chǎn)蛋性能高低和卵巢發(fā)育程度很重要。但FOXL2基因在不同時(shí)期的產(chǎn)蛋雞體內(nèi)各組織器官中的表達(dá)情況以及與蛋雞卵巢發(fā)育不同時(shí)期的相關(guān)性仍未見報(bào)道。本研究首先通過(guò)HE染色法觀察不同時(shí)期海蘭褐蛋雞卵巢的發(fā)育情況和組織學(xué)結(jié)構(gòu),并通過(guò)進(jìn)行卵泡數(shù)量的統(tǒng)計(jì)學(xué)分析,研究蛋雞卵巢卵泡的具體發(fā)育狀況。根據(jù)GenBank上收錄的紅原雞的FOXL2基因序列(登錄號(hào):JF_708868),通過(guò)生物信息學(xué)分析其保守序列及酶切位點(diǎn),利用Primmer_5.0設(shè)計(jì)并合成3對(duì)引物。通過(guò)瓊脂糖凝膠電泳試驗(yàn),篩選出最優(yōu)引物。采集30日齡,60日齡,90日齡,120日齡、160日齡、220日齡、330日齡、480日齡共8個(gè)時(shí)期的蛋雞的下丘腦、眼瞼、肌肉、心臟、胸腺、肝臟、脾臟、腎臟組織樣品,進(jìn)行qR T-PCR試驗(yàn)。對(duì)比不同時(shí)期卵巢中FOXL2基因相對(duì)表達(dá)量的差異,分析FOXL2基因與雞卵巢發(fā)育及其與產(chǎn)蛋性能的相關(guān)性;對(duì)比FOXL2基因在不同時(shí)期和不同組織中相對(duì)表達(dá)量的變化,分析FOXL2基因與蛋雞機(jī)體發(fā)育的相關(guān)性。HE染色和統(tǒng)計(jì)學(xué)分析結(jié)果表明,220日齡卵巢的體積和重量最大,之后卵巢逐漸萎縮,重量減少;隨日齡增加,總卵泡數(shù)逐漸減少,初級(jí)卵泡數(shù)呈現(xiàn)先增加后減少趨勢(shì),60日齡的初級(jí)卵泡數(shù)最高,然后逐漸減少;120日齡的次級(jí)卵泡數(shù)最高,并且明顯高于其他日齡產(chǎn)蛋雞;從30日齡開始,隨日齡增加,卵泡閉鎖率逐漸增多。qRT-PCR試驗(yàn)結(jié)果表明,120日齡雞卵巢中FOXL2基因的相對(duì)表達(dá)量高于其他日齡。FOXL2的表達(dá)與蛋雞卵巢發(fā)育存在相關(guān)性,產(chǎn)蛋雞不同組織中FOXL2基因相對(duì)表達(dá)量伴隨著產(chǎn)蛋雞組織器官的發(fā)育而變化,說(shuō)明FOXL2基因與產(chǎn)蛋雞其他組織的發(fā)育也存在一定的關(guān)系。本研究有助于進(jìn)一步認(rèn)識(shí)FOXL2基因在調(diào)控蛋雞卵巢發(fā)育和產(chǎn)蛋性能方面的作用,為深入研究該基因的生物學(xué)特性和功能奠定了基礎(chǔ)。
[Abstract]:The FOXL2 gene is the first sex-dimorphic marker associated with ovarian differentiation found in vertebrates and the first of the human autosomal genes to be thought to promote the early development of the human eyelid. An important factor in ovarian development and maintenance of ovarian function. With the deepening of related studies, it has been found that FOXL2 gene is expressed specifically in human eyelid, embryo and adult ovarian granulosa cells, and it is associated with premature ovarian failure, ovarian tumor, infertility. FOXL2 gene plays an important role in inhibiting follicular apoptosis and promoting ovarian reserve function. The normal expression of FOXL2 gene plays an important role in laying performance and ovarian development degree. However, the expression of FOXL2 gene in various tissues and organs of laying hens at different stages and the correlation with different stages of ovarian development of laying hens have not been reported. In this study, the development and histological structure of the ovary of Hailan brown laying hens at different stages were observed by HE staining method, and the specific development status of ovarian follicles was studied by statistical analysis of the number of follicles. Three pairs of primers were designed and synthesized by bioinformatics based on the sequence of FOXL2 gene (accession number: JFs 708868) collected on GenBank. Three pairs of primers were designed and synthesized by Primmer_5.0. The best primers were screened by agarose gel electrophoresis. The hypothalamus, eyelid, muscle, heart, thymus, liver, spleen and kidney of 8 laying hens were collected for QR T-PCR test in the hypothalamus, eyelid, muscle, heart, thymus, liver, spleen and kidney. To compare the relative expression of FOXL2 gene in different stages of ovary, to analyze the relationship between FOXL2 gene and ovarian development and egg laying performance, and to compare the changes of relative expression of FOXL2 gene in different stages and tissues. The correlation between FOXL2 gene and the development of laying hens was analyzed. The results of HE staining and statistical analysis showed that the size and weight of ovaries were the largest at the age of 220 days, then the ovaries gradually shrank and the weight decreased, and the total follicles decreased with the increase of age. The number of primary follicles increased first and then decreased. The number of primary follicles at 60 days of age was the highest, and the number of secondary follicles at 120 days of age was the highest, which was significantly higher than that of other laying hens, and increased with the age of 30 days. The results of follicular atresia. QRT-PCR test showed that the relative expression of FOXL2 gene in the ovaries of 120-day-old hens was higher than that of other day-old chickens, and there was a correlation between the expression of FOXL2 gene and the development of ovaries of laying hens. The relative expression of FOXL2 gene in different tissues of laying hens changed with the development of the tissues and organs of laying hens, which indicated that the FOXL2 gene had a certain relationship with the development of other tissues of laying hens. This study is helpful to further understand the role of FOXL2 gene in the regulation of ovarian development and egg laying performance of laying hens and lay a foundation for further study on the biological characteristics and functions of the gene.
【學(xué)位授予單位】：河南科技學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S831

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