辽宁绒山羊公羊精子发生与生精性能的研究
发布时间：2018-08-20 20:15
【摘要】：辽宁绒山羊是全世界产绒量最高,也是我国利用最为广泛的绒山羊品种资源。随着全国绒山羊产区良种化进程的加速,如何提高辽宁绒山羊种公羊的繁殖潜能与利用效率已为破解当前绒山羊业健康发展的瓶颈性问题,但解决该问题的关键是了解辽宁绒山羊公羊的精子发生及其随年龄增长所引发的精液品质变化的规律,及在此过程中睾丸生殖细胞是如何变化等。为此,本研究分别以不同年龄的辽宁绒山羊为研究对象,通过对辽宁绒山羊公羔睾丸等主要生殖器官发育及精子发生进、对调控细胞凋亡的Bcl-2基因和Bax基因的蛋白与转录水平表达以及辽宁绒山羊精液品质及其利用进行了系统的研究。1.通过对40只出生日期相接近的公羔0~180日龄间的连续观测,获得如下研究结果:(1)辽宁绒山羊公羔的性行为发育较早,各个性行为在断奶后15~30d内全部出现并完善。(2)辽宁绒山羊公羔在出生后生长发育极为迅速,生后一个月龄时的体重为初生时的3倍以上,而断奶后至6月龄体重增加了近1倍左右。(3)辽宁绒山羊公羔的主要生殖器官均呈现随日龄增长而迅速发育的趋势,6月龄时单侧睾丸、单侧附睾、阴茎长度和精细管直径分别为初生时的238倍、53倍、7.2倍和6.14倍,以单侧睾丸的重量生长发育最为迅速。(4)辽宁绒山羊公羔睾丸内的精原细胞数量自出生后呈逐渐增加的状态,30日龄时出现了初级精母细胞,60日龄时出现次级精母细胞,90日龄时出现精子细胞和精子,180日龄时精子细胞数量开始减少而精子开始增加;辽宁绒山羊公羔在4月龄可以采集出精液,但精液品质中的各个指标均低,尚不可进行受精,但5月龄之后则其精液品质指标已经与成年公羊相接近。(5)随着辽宁绒山羊公羔日龄的增长,其外周血清中睾酮的分泌水平也呈逐渐增加状态,30日龄以后的分泌水平分为为初生时的2.23倍、5倍、7.94倍、10.41倍、13.45倍和16.16倍。说明公羊的性行为决定公羊的种用性能和使用方向与年限;公羊初配与年龄和体重有关;睾丸的重要机能就是产生精子和分泌睾酮。2.通过对辽宁绒山羊8月龄~18月龄、18月龄~24月龄、36月龄以上不同年龄公羊在春夏秋冬4个季节内的精液物理性状检测,获得如下研究结果:各个年龄段的公羊的采精量和精子活力均是秋季优于春季、冬季,春季和冬季优于夏季,幼龄公羊的精子活力优于成年公羊;不同年龄阶段的精子密度均随季节发生变化,精子密度以秋季为最高;辽宁绒山羊成年公羊的顶体完整率以秋季为最高,而精子畸形率为最低,夏季公羊的顶体完整率最低而精子畸形率却最高;经典的卵黄稀释液稀释和无动物源性稀释液稀释法一样,在低温保存状态下,精液稀释倍数越大,精子体外存活的时间越长;温度越高,两种稀释方法在同一倍数下,精子的体外存活时间在逐渐减少。经典的卵黄稀释液稀释法,在同一温度下,精子在体外的存活时间,随着稀释倍数的增加而逐渐延长;但无动物源性稀释液稀释法,在同一温度下,精子在体外的存活时间,则随着稀释倍数的增加而逐渐减少。说明,幼龄公羊的精子活力优于成年羊,应选取幼龄公羊作为种公羊且最佳配种季节为秋季,可获得较高的受胎率。3.采用荧光定量PCR法对2月龄和15月龄辽宁绒山羊公羊睾丸的Bax和Bcl-2基因的mRNA表达的差异进行研究,获得如下研究结果:在辽宁绒山羊睾丸性成熟前后,Bax基因和Bcl-2基因的mRNA均有表达,Bax基因mRNA的表达量差异极显著(P0.01),Bax基因性成熟后的表达量是性成熟前的120倍,Bcl-2基因的mRNA的表达量差异不显著(P0.05),Bcl-2基因性成熟后的表达量是性成熟前的1.4倍,说明性成熟前后均有细胞凋亡现象,但以性成熟后为主。
[Abstract]:Liaoning cashmere goat is the highest cashmere production in the world and the most widely used cashmere goat breed resource in China. With the acceleration of the process of fine breeding in the cashmere goat producing areas of China, how to improve the reproductive potential and utilization efficiency of Liaoning cashmere goat breed ram has become a bottleneck problem to solve the current healthy development of cashmere goat industry, but to solve this problem. The key is to understand the spermatogenesis of Liaoning cashmere goats and the changes of sperm quality with age, and the changes of testicular germ cells in the process. The protein and transcription levels of Bcl-2 and Bax genes regulating apoptosis and the semen quality and utilization of Liaoning cashmere goats were studied systematically during breeding and spermatogenesis. The development of sexual behavior was earlier, and all sexual behaviors appeared and perfected within 15-30 days after weaning. (2) Liaoning cashmere goat male lambs grew very fast after birth. The weight at the age of one month after birth was more than three times that at the time of birth, and the weight increased nearly twice from weaning to 6 months. (3) The main reproductive organs of Liaoning cashmere goat male lambs were all present. At the age of 6 months, unilateral testis, unilateral epididymis, penis length and seminiferous tubule diameter were 238, 53, 7.2 and 6.14 times respectively. The weight of unilateral testis grew and developed most rapidly. (4) The number of spermatogonia in the testis of Liaoning cashmere goats increased gradually after birth. Primary spermatocytes appeared at 30 days of age, secondary spermatocytes appeared at 60 days of age, sperm cells and sperm appeared at 90 days of age, sperm cells began to decrease at 180 days of age and sperm began to increase; Liaoning cashmere goat male lambs could collect sperm at 4 months of age, but each index of semen quality was low and could not be fertilized yet, but the number of sperm cells and sperm began to decrease at 50 days of age. The semen quality index of Liaoning cashmere goats was similar to that of adult rams after one month of age. (5) With the increase of day-old Liaoning cashmere goats, the level of serum testosterone secretion increased gradually. After 30 days of age, the level of serum testosterone secretion was divided into 2.23 times, 5 times, 7.94 times, 10.41 times, 13.45 times and 16.16 times of that at birth. In order to determine the breeding performance and use direction of rams and years; ram primary mating is related to age and weight; the important function of testis is sperm production and testosterone secretion. 2. Through the Liaoning cashmere goats 8 months to 18 months old, 18 months to 24 months old, 36 months old rams of different ages in the spring, summer, autumn and winter four seasons physical characteristics of semen were detected. The results showed that the sperm collection and sperm motility of rams of all ages were better in autumn than in spring, winter, spring and winter were better than in summer, and the sperm motility of young rams was better than that of adult rams. The highest rate of acrosome integrity was in autumn, and the lowest rate of sperm deformity was in summer. The lowest rate of acrosome integrity and the highest rate of sperm deformity were in summer rams. The survival time of spermatozoa in vitro decreased gradually with the same dilution ratio. The survival time of spermatozoa in vitro was gradually prolonged with the increase of dilution ratio in the classical yolk dilution method at the same temperature, but the survival time of spermatozoa in vitro was prolonged with the increase of dilution ratio in the non-animal dilution method at the same temperature. The results showed that the sperm motility of young ram was better than that of adult ram, and the best breeding season was autumn. 3. The difference of Bax and Bcl-2 mRNA expression between 2-month-old and 15-month-old Liaoning cashmere goat testes was detected by fluorescence quantitative PCR. The results showed that Bax gene and Bcl-2 gene mRNA were expressed before and after sexual maturation in Liaoning cashmere goats. The expression of Bax gene mRNA was significantly different (P 0.01). The expression of Bax gene after sexual maturation was 120 times higher than that before sexual maturation. The expression of Bcl-2 gene mRNA was not significantly different (P 0.05). The expression level after maturation was 1.4 times higher than that before sexual maturation, indicating that apoptosis occurred before and after sexual maturation, but mainly after sexual maturation.
【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S827
本文编号：2194864
[Abstract]:Liaoning cashmere goat is the highest cashmere production in the world and the most widely used cashmere goat breed resource in China. With the acceleration of the process of fine breeding in the cashmere goat producing areas of China, how to improve the reproductive potential and utilization efficiency of Liaoning cashmere goat breed ram has become a bottleneck problem to solve the current healthy development of cashmere goat industry, but to solve this problem. The key is to understand the spermatogenesis of Liaoning cashmere goats and the changes of sperm quality with age, and the changes of testicular germ cells in the process. The protein and transcription levels of Bcl-2 and Bax genes regulating apoptosis and the semen quality and utilization of Liaoning cashmere goats were studied systematically during breeding and spermatogenesis. The development of sexual behavior was earlier, and all sexual behaviors appeared and perfected within 15-30 days after weaning. (2) Liaoning cashmere goat male lambs grew very fast after birth. The weight at the age of one month after birth was more than three times that at the time of birth, and the weight increased nearly twice from weaning to 6 months. (3) The main reproductive organs of Liaoning cashmere goat male lambs were all present. At the age of 6 months, unilateral testis, unilateral epididymis, penis length and seminiferous tubule diameter were 238, 53, 7.2 and 6.14 times respectively. The weight of unilateral testis grew and developed most rapidly. (4) The number of spermatogonia in the testis of Liaoning cashmere goats increased gradually after birth. Primary spermatocytes appeared at 30 days of age, secondary spermatocytes appeared at 60 days of age, sperm cells and sperm appeared at 90 days of age, sperm cells began to decrease at 180 days of age and sperm began to increase; Liaoning cashmere goat male lambs could collect sperm at 4 months of age, but each index of semen quality was low and could not be fertilized yet, but the number of sperm cells and sperm began to decrease at 50 days of age. The semen quality index of Liaoning cashmere goats was similar to that of adult rams after one month of age. (5) With the increase of day-old Liaoning cashmere goats, the level of serum testosterone secretion increased gradually. After 30 days of age, the level of serum testosterone secretion was divided into 2.23 times, 5 times, 7.94 times, 10.41 times, 13.45 times and 16.16 times of that at birth. In order to determine the breeding performance and use direction of rams and years; ram primary mating is related to age and weight; the important function of testis is sperm production and testosterone secretion. 2. Through the Liaoning cashmere goats 8 months to 18 months old, 18 months to 24 months old, 36 months old rams of different ages in the spring, summer, autumn and winter four seasons physical characteristics of semen were detected. The results showed that the sperm collection and sperm motility of rams of all ages were better in autumn than in spring, winter, spring and winter were better than in summer, and the sperm motility of young rams was better than that of adult rams. The highest rate of acrosome integrity was in autumn, and the lowest rate of sperm deformity was in summer. The lowest rate of acrosome integrity and the highest rate of sperm deformity were in summer rams. The survival time of spermatozoa in vitro decreased gradually with the same dilution ratio. The survival time of spermatozoa in vitro was gradually prolonged with the increase of dilution ratio in the classical yolk dilution method at the same temperature, but the survival time of spermatozoa in vitro was prolonged with the increase of dilution ratio in the non-animal dilution method at the same temperature. The results showed that the sperm motility of young ram was better than that of adult ram, and the best breeding season was autumn. 3. The difference of Bax and Bcl-2 mRNA expression between 2-month-old and 15-month-old Liaoning cashmere goat testes was detected by fluorescence quantitative PCR. The results showed that Bax gene and Bcl-2 gene mRNA were expressed before and after sexual maturation in Liaoning cashmere goats. The expression of Bax gene mRNA was significantly different (P 0.01). The expression of Bax gene after sexual maturation was 120 times higher than that before sexual maturation. The expression of Bcl-2 gene mRNA was not significantly different (P 0.05). The expression level after maturation was 1.4 times higher than that before sexual maturation, indicating that apoptosis occurred before and after sexual maturation, but mainly after sexual maturation.
【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S827
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