GnIH对母鹅繁殖性能和公鸡生殖发育的影响

发布时间：2018-02-27 13:51

本文关键词： 马岗鹅公鸡促性腺抑制激素繁殖性能生殖发育　出处：《仲恺农业工程学院》2015年硕士论文　论文类型：学位论文

【摘要】：Gn IH可以作用于垂体前叶或作用于Gn RH来抑制促性腺激素的合成与释放,是调节动物繁殖性能的重要生殖激素之一。目前用生殖免疫调控的方法提高家畜繁殖力已受到广泛关注。许多研究应用抑制素、催乳素、Gn IH等生殖激素作用于动物体内或体外实验均可发现其参与调控动物的生长繁殖。因此,本实验主要以马岗鹅和公鸡为研究对象,从外源性给予Gn IH和主动免疫Gn IH两种不同的研究方法进行研究:实验一选取健康的成年母鹅36只,平均分成3组。第一组仅在d1给予30%PVP-K30溶解的Gn IH溶液,给予Gn IH 1 mg/只;第二组在d1、d4、d7给予生理盐水溶解的Gn IH溶液,给予Gn IH 1 mg/只;第三组在d1、d4、d7给予生理盐水,给予1 m L每只。试验中记录母鹅每日产蛋数和就巢情况;并在d1、d4、d7、d10定时采集血清,在试验结束时屠宰母鹅,采集下丘脑和垂体组织样和检查卵泡发育情况。研究表明给予产蛋母鹅外源性Gn IH能一定程度上抑制自身Gn IH的基因表达,降低血液中LH浓度,提高雌二醇浓度,主要通过抑制中小卵泡发育,进而在一定程度上抑制母鹅产蛋。实验二选取30只处于就巢期健康的成年马岗鹅母鹅,平均分成免疫组和对照组(n=15),试验在正式开始时,对母鹅进行人工强制终止就巢。免疫组分别在d1、d21、d41接种含Gn IH蛋白的疫苗1 m L/只(以此含1.0、1.0、0.5 mg的Gn IH蛋白),对照组在d1、d21、d41注射未含免疫原的生理盐水制备的乳化苗。两组鹅试验开始饲养于短光照(11L:13D)下以继续维持正常产蛋,在d72将每天的光照时间延长至18小时(18L:6D)至d95试验结束。试验期记录鹅的就巢发生、每日产蛋数,并每20天对母鹅采血一次,分离血清留待激素测定。在试验结束时对两组母鹅进行屠宰,检查卵泡发育情况,并采集下丘脑和垂体样本,留待基因表达水平检测。研究结果表明对成年母鹅主动免疫Gn IH可诱导体内产生特异性抗Gn IH的抗体,抑制Gn IH的基因表达,略微提高了Gn RH、VIP和PRL的基因表达,调节血清中LH、E2、P4等相关生殖激素进而提高母鹅的产蛋性能。另外,延长光照会抑制母鹅的产蛋,但其对对照组的作用比免疫组更强,这可能是因为主动免疫Gn IH对母鹅的产蛋促进作用还在维持。实验三选120只60日龄健康公鸡,随机平均分成两组,免疫组和对照组,所有鸡均采用笼养,每笼2只。免疫组接种含Gn IH蛋白的疫苗1 m L/只(含0.5 mg的Gn IH蛋白),对照组注射未含免疫原的生理盐水制备的乳化苗。分别在试验的d1、d21和d41(即60、80和100日龄)进行胸部肌肉注射接种疫苗。在试验期,从60日龄开始,每隔20天进行对公鸡进行采血,所得血清保存于-20℃,留待测定血液中抗体水平和激素水平;每隔30天分别从两组随机挑选8只公鸡进行屠宰,采取睾丸组织并检测其发育情况,并采集下丘脑和垂体留待基因表达水平检测。从195日龄开始两组公鸡进行人工采精训练(按摩法),待一周建立条件反射后,每隔两天采精一次进行精液品质检测。研究结果表明,对发育期公鸡主动免疫Gn IH能有效的刺激了机体产生特异性抗Gn IH抗体,并在整个实验过程中维持较高水平。免疫Gn IH后能抑制Gn IH、VIP、PRL的基因表达,提高LH的基因表达,进而促进睾丸发育和血液循环中睾酮、INH、ACT的分泌,提高精液品质。
[Abstract]:Gn IH can function in the anterior pituitary function in Gn or RH to inhibit the synthesis and release of gonadotropin, is one of the most important reproductive hormone regulation of animal reproductive performance. The current method for reproductive immune regulation to improve the fecundity of cattle has attracted widespread attention. Many studies by prolactin inhibin, Gn, IH and other reproductive hormones in animal in vivo or in vitro experiments can be found which is involved in the regulation of animal growth and reproduction. Therefore, this experiment mainly in Magang geese and cock as the research object, Gn IH and Gn IH active immunization with two different research methods from exogenous research: the first experiment healthy adult female geese 36, divided into 3 groups the first group was given 30%PVP-K30 dissolved in D1 Gn IH Gn IH mg/ 1 solution, given only; second in D1 group, D4 Gn, IH D7 solution of saline solution, given the Gn IH 1 mg/; third in D1 group, D4, D7 Saline, 1 m L each. The goose egg number per Nissan and nest record test; and in D1, D4, D7, D10 in serum collected at the end of the experiment, the slaughter of the goose, collecting the hypothalamus and pituitary tissue samples and check the follicular development. The results show that the goose egg exogenous Gn IH can inhibit the Gn gene expression of IH, reduce the LH concentration in the blood, improve the concentration of estradiol, mainly through the inhibition of small follicular development, thereby inhibiting the goose egg in a certain extent. In experiment two, a total of 30 healthy adult in broodiness period Magang goose goose, divided into immune group and control group (n=15), the test officially started, the goose was forced to terminate the broodiness. Artificial immune group respectively in D1, D21, d41 Gn IH protein vaccine inoculation with 1 m L/ only (mg Gn to 1.0,1.0,0.5 containing IH protein), the control group in D1, D21, d41 without injection Immunogenicity of saline prepared emulsion seedlings. Two groups of goose started to short photoperiod test (11L:13D) to continue to maintain normal production, the D72 daily illumination time is extended to 18 hours (18L:6D d95) to the end of the experiment. Recorded during the experiment of goose nest, daily egg number, and every 20 days for the goose blood once, for the separation and determination of serum hormone. The slaughter of two groups of goose at the end of the test, check the follicular development and acquisition of the hypothalamus and pituitary samples, for the detection of expression level of genes. The results of the study show that the antibody of adult goose Gn active immunization can induce the body to produce specific IH anti Gn IH, inhibit the expression of Gn IH gene, and slightly increased the Gn RH expression of VIP and PRL genes, regulating the serum levels of LH, E2, P4 and other related reproductive hormones and improve the performance of laying goose. In addition, extended light inhibition goose egg, but the The role of the two groups were stronger, this may be because the active immunization of Gn IH on the goose egg role still maintain. Experiment three choose 120 healthy 60 day old cocks were randomly divided into two groups, immune group and control group, all the chickens were used in cages, each cage 2 group. Immune vaccination Gn containing IH protein vaccine M 1 L/ (0.5 mg Gn IH protein), the control group was injected with saline without immunogen prepared emulsion vaccine. Respectively in the test of D1, D21 and d41 (60,80 and 100 days of age) chest muscle injection vaccination. In the trial period. From the beginning of the 60 days, every 20 days of blood serum of cock, stored in the -20 the C, determination of antibody levels and hormone levels in the blood for; every 30 days respectively from two randomly selected group of 8 cocks were slaughtered, take testicular tissue and detect its development situation, and the hypothalamic and pituitary for acquisition Gene expression detection. From 195 days of age two group cock artificial sperm collection training (massage), for a week after the establishment of conditioned reflex, once every two days of semen collection semen quality testing. The results show that can effectively stimulate the body to produce specific anti Gn IH antibody on the development period of the cock active immunization of Gn IH, and to maintain a high level during the whole experiment. Gn IH Gn can inhibit the immune IH, VIP, PRL gene expression, enhance the expression of LH gene, and promote testosterone, testis development and blood circulation in INH, ACT secretion, improve semen quality.

【学位授予单位】：仲恺农业工程学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S831;S835

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