本文關(guān)鍵詞： FSHR基因 KISS1基因 轉(zhuǎn)基因小鼠 產(chǎn)仔數(shù) 陽性率 豬　出處：《西南大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：繁殖性能是家畜的重要經(jīng)濟(jì)性狀,繁殖力低是限制現(xiàn)代畜牧業(yè)發(fā)展的原因之一。卵泡刺激素受體(Follicle Stimulating Hormone Receptor, FSHR)和親吻素(KISS1)基因?qū)倚蟮漠a(chǎn)仔數(shù)有顯著影響。卵泡刺激素(Follicle stimulating hormone,FSH)在促進(jìn)和維持性腺發(fā)育和生殖功能中發(fā)揮重要作用,但是其生理作用需通過分布于細(xì)胞膜表面的卵泡刺激素受體(Follicle stimulating hormone receptor, FSHR)介導(dǎo)。KISS1基因在下丘腦-垂體-性腺軸系統(tǒng)中起到中樞節(jié)點(diǎn)作用。KISS1基因的翻譯產(chǎn)物kisspeptins刺激GnRH分泌,激活性腺軸,影響黃體生成素(Luteinizing hormone,LH)和FSH的分泌,進(jìn)而對(duì)哺乳動(dòng)物生殖內(nèi)分泌起調(diào)控作用。KISS1及其受體G蛋白偶聯(lián)受體(G protein-coupled receptor 54,GPR54)基因還是調(diào)節(jié)動(dòng)物初情期的關(guān)鍵基因。實(shí)驗(yàn)室前期通過分別構(gòu)建小鼠卵巢顆粒細(xì)胞特異表達(dá)的豬FSHR基因(pFSHR)真核載體AMH-pFSHR和小鼠腦部特異表達(dá)豬KISS1基因(pKISS1)的真核載體pCaMKIIa-KISS1,通過顯微注射的方法制備了轉(zhuǎn)豬FSHR和KISS1基因小鼠。本文針對(duì)F1-F5代小鼠陽性率、pFSHR和pKISS1基因在小鼠體內(nèi)的表達(dá)情況、以及對(duì)雌性小鼠繁殖性能的影響進(jìn)行研究,實(shí)驗(yàn)結(jié)果如下：1.轉(zhuǎn)豬FSHR基因小鼠：F1-F5代轉(zhuǎn)基因小鼠平均每窩產(chǎn)仔數(shù)均高于野生型小鼠,F1和F3代的窩產(chǎn)仔數(shù)達(dá)到顯著水平(P0.05)在轉(zhuǎn)基因小鼠卵巢中特異表達(dá),心臟、肝臟、脾臟、腎臟、子宮及下丘腦中則未檢測(cè)到其表達(dá),而在轉(zhuǎn)基因小鼠和野生型小鼠的卵巢都有mFSHR mRNA的表達(dá)。豬FSHR蛋白在轉(zhuǎn)基因小鼠卵巢中有表達(dá),其表達(dá)水平在陽性小鼠之間沒有顯著差異,與mRNA水平結(jié)果一致。FSH、LH、雌二醇、孕酮在間情期轉(zhuǎn)基因小鼠和野生型小鼠中沒有顯著差異(P0.05)。產(chǎn)仔數(shù)曲線表明轉(zhuǎn)基因小鼠和野生型小鼠的窩產(chǎn)仔數(shù)隨著年齡增加先升高,再降低；在同一鼠齡,轉(zhuǎn)基因小鼠的產(chǎn)仔數(shù)比野生型小鼠多,且都在220日齡左右具有最大的窩產(chǎn)仔數(shù),且達(dá)到顯著水平(P0.05)；轉(zhuǎn)基因小鼠的生產(chǎn)間隔時(shí)間比野生型小鼠更短(P0.05)。2.轉(zhuǎn)豬KISS1基因小鼠： F1-F5代轉(zhuǎn)基因小鼠平均每窩產(chǎn)仔數(shù)均高于野生型小鼠,而且F1和F2代的窩產(chǎn)仔數(shù)達(dá)到顯著水平(分別為P0.01和P0.05)。pKISS1 mRNA在轉(zhuǎn)基因小鼠下丘腦中有表達(dá),其他組織中則未檢測(cè)到其表達(dá)；在轉(zhuǎn)基因小鼠和野生型小鼠的卵巢和下丘腦中都能檢測(cè)到mKISS1 mRNA的表達(dá)。豬kisspeptin蛋白在轉(zhuǎn)基因小鼠下丘腦中有表達(dá)。F4代轉(zhuǎn)基因小鼠血清中FSH和雌二醇激素濃度顯著高于野生型小鼠(P0.05)。產(chǎn)仔數(shù)曲線表明：同一鼠齡轉(zhuǎn)基因小鼠比野生型小鼠具有更多的產(chǎn)仔數(shù),都在200日齡左右具有最大的窩產(chǎn)仔數(shù),且生產(chǎn)間隔時(shí)間比野生型小鼠更短。F4代轉(zhuǎn)豬KISS1基因小鼠后代提前進(jìn)入初情期,卵巢重量顯著增加(P0.05)。
[Abstract]:Reproductive performance is an important economic trait of livestock. Low fecundity is one of the reasons that restrict the development of modern animal husbandry. Follicle Stimulating Hormone Receptor. FSHRand KISS1) genes have a significant effect on litter size of domestic animals. Follicle stimulating hormone. FSHs play an important role in promoting and maintaining gonadal development and reproductive function. However, its physiological function is through Follicle stimulating hormone receptor, which is distributed on the surface of the cell membrane. FSHR). KISS1 gene plays a central node role in hypothalamus-pituitary-gonadal axis system. The translation product of KISS1 gene kisspeptins stimulates the secretion of GnRH. The activation of gonadal axis affects the secretion of luteinizing hormone (LHH) and FSH. KISS1 and its receptor G protein coupled receptor G protein-coupled receptor 54. GPR54) gene is also the key gene to regulate the early estrus of animal. The porcine FSHR gene expressed specifically in granulosa cells of mouse ovary was constructed by constructing porcine FSHR gene pFSHRs in the early stage of laboratory. Eukaryotic vector AMH-pFSHR and mouse brain specific eukaryotic vector pCaMKIIa-KISS1 expressing porcine KISS1 gene pKISS1). The transgenic FSHR and KISS1 gene mice were prepared by microinjection. The expression of pFSHR and pKISS1 genes in mice with positive rate of F1-F5 generation was studied. The results were as follows: 1. The average litter size of transgenic mice with FSHR gene was higher than that of wild-type mice. The litter size of F1 and F3 generation reached a significant level (P0.05), but no expression was found in the ovary, heart, liver, spleen, kidney, uterus and hypothalamus of transgenic mice. The expression of mFSHR mRNA was found in the ovaries of transgenic mice and wild-type mice, and porcine FSHR protein was expressed in the ovaries of transgenic mice. There was no significant difference between the positive mice and the positive mice, and the results were consistent with the results of mRNA. There was no significant difference in progesterone between transgenic mice and wild type mice. The litter size curve showed that the litter size of transgenic mice and wild type mice increased first and then decreased with age. The litter size of transgenic mice was more than that of wild-type mice at the same age, and it had the largest litter size at 220 days old, and reached the significant level of P0.05; The production interval of transgenic mice was shorter than that of wild-type mice. The average litter size of transgenic mice of F1-F5 generation was higher than that of wild-type mice. Moreover, the litter size of F1 and F2 generation reached a significant level (P0.01 and P0.05, respectively. PKISS1 mRNA was expressed in the hypothalamus of transgenic mice. No expression was detected in other tissues. MKISS1 was detected in ovary and hypothalamus of transgenic mice and wild-type mice. Expression of mRNA. The expression of porcine kisspeptin protein in the hypothalamus of transgenic mice. F4 generation transgenic mice serum FSH and estradiol hormone concentrations were significantly higher than wild-type mice (. The curve of litter size showed that transgenic mice of the same age had more offspring than wild-type mice. All of them had the largest litter size at the age of 200 days, and the interval of production was shorter than that of wild type mice. The progenies of transgenic pig KISS1 gene mice of passage F4 entered the early stage of estrus earlier than those of wild type mice. The weight of ovary increased significantly (P 0.05).
【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S828

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相關(guān)期刊論文 前1條

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本文編號(hào)：1442044