能蛋水平对北京油鸡后期繁殖性能的影响及睾丸基因表达谱的建立
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摘要
本研究是以50-58周龄北京油鸡种公鸡为试验样本,探讨地方种公鸡繁殖后期对精液品质的影响最佳的日粮能量、蛋白水平,并通过孵化试验进行进一步的验证。为更深入地研究影响精液品质的分子机理,本试验首次采用数字基因表达谱技术(DGE)对种公鸡无精睾丸和有精睾丸RNA样构建表达谱,对差异表达的基因进行初步的分析。
试验一日粮能量、蛋白水平对种公鸡后期繁殖性能的影响。将99只50周龄北京油鸡种公鸡随机分为9个处理,每个处理11只鸡,分别饲喂能量(11.30MJ/kg、11.72 MJ/kg、12.14 MJ/kg)和蛋白(12%、14%、16%)3×3因子组合的9种日粮。试验结果显示,蛋白水平对生长性能和繁殖性能无显著影响(P>0.05),但精子活力随蛋白升高有降低的趋势;能量为12.14 MJ/kg时对采食量和精子活率有显著性影响(P<0.05);11.30MJ/kg-12%组的畸形率显著高于其它组(P<0.05);能量和蛋白之间有较强的互作,孵化验证试验显示11.72 MJ/kg-12%组的受精率、孵化率和健雏率较佳。北京油鸡种公鸡后期日粮蛋白水平为12%,能量水平为11.72 MJ / kg。
试验二应用数字基因表达谱技术(DGE)建立种公鸡无精症睾丸基因表达谱,初步分析差异基因的表达和功能。本试验筛选一只不产精种公鸡和一只精液正常的种公鸡睾丸样本,提取睾丸RNA,建立表达谱,分析无精症和正常种公鸡睾丸基因的表达情况。结果显示,无精症睾丸和正常睾丸共有16379个基因存在表达差异,上调基因545个,下调基因202个。差异表达基因主要参与烟酸和烟酰胺的代谢、造血干细胞生成、细胞生成、细胞周期、细胞结构、Wnt信号通路、炎症反应等生物学通路。在正常睾丸样中与精子发生、蛋白代谢、线粒体结构、呼吸链组分和影响多种催化酶活性的基因高表达,这可能是精子正常生成的原因。在无精睾丸中与炎症反应、负向调控蛋白激酶、异常代谢、蛋白酶抑制剂活性等相关基因高表达,这些因素可能是导致精子生成障碍的原因。建立种公鸡无精症表达谱为进一步分析精子生成的分子机制和探究无精症的病因将会有所帮助。
This study chose 50-58 week Beijing-You chickens as test samples to explore that effects of energy-protein levels on reproduction performance in late reproductive period. It was also further validated by incubation tests. To further study the molecular mechanism of sperm quality, the trial first used digital gene expression profiling technology (DGE) of the species of cock testis and azoospermia testicular sperm RNA samples to construct expression profiles and analyse differential expressed genes.
Test 1. This experiment was conducted to study the effect of energy-protein levels on reproduction performance in late reproductive period of Beijing-You Chickens. 99 50-week old Beijing-You chickens were assigned to 9 treatments, 11 cokes every treatment. In a 3×3 factorial arrangem to experimental diets with11.30、11.72、12.14MJ/kg ME; each contained 12% ,14% and16% CP. The results showed that protein levels on growth performance and reproductive performance was not significantly affected(P<0.05). But sperm motility tended to be improved with the lower of CP level; 12.14MJ/kg energy level on feed intake and sperm motility rate were significantly affected(P<0.05). 11.30MJ/kg-12% group of deformity rate was significantly higher than other groups. We obtained this result by incubation verification tests.Fertilization rate, hatching rate and healthy chick rate of the 11.72 MJ/kg-12% group was the best. The results show that Beijing-You chicken dietary CP is 12%, ME is 11.72 MJ/kg in late reproductive period is suggested.
Test 2. The experiment used the application of digital gene expression profiling technology (DGE) to establish species of cock testis gene expression azoospermia, preliminarily analysing differences in gene expression and functional analysis. The experiment screened samples of a rooster which does not produce fine sperm and a rooster testicles semen of the normal species and extracted testicular RNA to establish expression profilesand analyse azoospermia and normal kinds of cock testis gene expression. The results show that testicular azoospermia and normal testicular expression have a total of 16379 different genes, 545 up-regulated genes and 202 down-regulated genes. Differentially expressed genes is mainly involved in nicotinic acid, niacinamid the metabolism, hematopoietic stem cell formation,cell formation, the cell cycle, the cellular structure Wnt signaling pathways and inflammatory reaction biological pathways.In normal testicular sperm sample with occurrence, protein metabolism, mitochondrial structure, composition and respiratory chain of catalytic influence of gene expression of enzyme activity. This may be reasons that high sperm normal generated. In no refined testis and inflammation, negative control protein kinase, abnormal metabolism, protease inhibitors activity and other related gene high expression, these factors maybe cause the sperm production barrier. Establishing a rooster no refined disease would be helpful, for further analysis express spectrum of sperm production molecular mechanism and study the etiopathogenisis of azoospermia.
试验一日粮能量、蛋白水平对种公鸡后期繁殖性能的影响。将99只50周龄北京油鸡种公鸡随机分为9个处理,每个处理11只鸡,分别饲喂能量(11.30MJ/kg、11.72 MJ/kg、12.14 MJ/kg)和蛋白(12%、14%、16%)3×3因子组合的9种日粮。试验结果显示,蛋白水平对生长性能和繁殖性能无显著影响(P>0.05),但精子活力随蛋白升高有降低的趋势;能量为12.14 MJ/kg时对采食量和精子活率有显著性影响(P<0.05);11.30MJ/kg-12%组的畸形率显著高于其它组(P<0.05);能量和蛋白之间有较强的互作,孵化验证试验显示11.72 MJ/kg-12%组的受精率、孵化率和健雏率较佳。北京油鸡种公鸡后期日粮蛋白水平为12%,能量水平为11.72 MJ / kg。
试验二应用数字基因表达谱技术(DGE)建立种公鸡无精症睾丸基因表达谱,初步分析差异基因的表达和功能。本试验筛选一只不产精种公鸡和一只精液正常的种公鸡睾丸样本,提取睾丸RNA,建立表达谱,分析无精症和正常种公鸡睾丸基因的表达情况。结果显示,无精症睾丸和正常睾丸共有16379个基因存在表达差异,上调基因545个,下调基因202个。差异表达基因主要参与烟酸和烟酰胺的代谢、造血干细胞生成、细胞生成、细胞周期、细胞结构、Wnt信号通路、炎症反应等生物学通路。在正常睾丸样中与精子发生、蛋白代谢、线粒体结构、呼吸链组分和影响多种催化酶活性的基因高表达,这可能是精子正常生成的原因。在无精睾丸中与炎症反应、负向调控蛋白激酶、异常代谢、蛋白酶抑制剂活性等相关基因高表达,这些因素可能是导致精子生成障碍的原因。建立种公鸡无精症表达谱为进一步分析精子生成的分子机制和探究无精症的病因将会有所帮助。
This study chose 50-58 week Beijing-You chickens as test samples to explore that effects of energy-protein levels on reproduction performance in late reproductive period. It was also further validated by incubation tests. To further study the molecular mechanism of sperm quality, the trial first used digital gene expression profiling technology (DGE) of the species of cock testis and azoospermia testicular sperm RNA samples to construct expression profiles and analyse differential expressed genes.
Test 1. This experiment was conducted to study the effect of energy-protein levels on reproduction performance in late reproductive period of Beijing-You Chickens. 99 50-week old Beijing-You chickens were assigned to 9 treatments, 11 cokes every treatment. In a 3×3 factorial arrangem to experimental diets with11.30、11.72、12.14MJ/kg ME; each contained 12% ,14% and16% CP. The results showed that protein levels on growth performance and reproductive performance was not significantly affected(P<0.05). But sperm motility tended to be improved with the lower of CP level; 12.14MJ/kg energy level on feed intake and sperm motility rate were significantly affected(P<0.05). 11.30MJ/kg-12% group of deformity rate was significantly higher than other groups. We obtained this result by incubation verification tests.Fertilization rate, hatching rate and healthy chick rate of the 11.72 MJ/kg-12% group was the best. The results show that Beijing-You chicken dietary CP is 12%, ME is 11.72 MJ/kg in late reproductive period is suggested.
Test 2. The experiment used the application of digital gene expression profiling technology (DGE) to establish species of cock testis gene expression azoospermia, preliminarily analysing differences in gene expression and functional analysis. The experiment screened samples of a rooster which does not produce fine sperm and a rooster testicles semen of the normal species and extracted testicular RNA to establish expression profilesand analyse azoospermia and normal kinds of cock testis gene expression. The results show that testicular azoospermia and normal testicular expression have a total of 16379 different genes, 545 up-regulated genes and 202 down-regulated genes. Differentially expressed genes is mainly involved in nicotinic acid, niacinamid the metabolism, hematopoietic stem cell formation,cell formation, the cell cycle, the cellular structure Wnt signaling pathways and inflammatory reaction biological pathways.In normal testicular sperm sample with occurrence, protein metabolism, mitochondrial structure, composition and respiratory chain of catalytic influence of gene expression of enzyme activity. This may be reasons that high sperm normal generated. In no refined testis and inflammation, negative control protein kinase, abnormal metabolism, protease inhibitors activity and other related gene high expression, these factors maybe cause the sperm production barrier. Establishing a rooster no refined disease would be helpful, for further analysis express spectrum of sperm production molecular mechanism and study the etiopathogenisis of azoospermia.
引文
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