【摘要】：遼寧絨山羊是全世界產(chǎn)絨量最高,也是我國利用最為廣泛的絨山羊品種資源。隨著全國絨山羊產(chǎn)區(qū)良種化進(jìn)程的加速,如何提高遼寧絨山羊種公羊的繁殖潛能與利用效率已為破解當(dāng)前絨山羊業(yè)健康發(fā)展的瓶頸性問題,但解決該問題的關(guān)鍵是了解遼寧絨山羊公羊的精子發(fā)生及其隨年齡增長所引發(fā)的精液品質(zhì)變化的規(guī)律,及在此過程中睪丸生殖細(xì)胞是如何變化等。為此,本研究分別以不同年齡的遼寧絨山羊為研究對象,通過對遼寧絨山羊公羔睪丸等主要生殖器官發(fā)育及精子發(fā)生進(jìn)、對調(diào)控細(xì)胞凋亡的Bcl-2基因和Bax基因的蛋白與轉(zhuǎn)錄水平表達(dá)以及遼寧絨山羊精液品質(zhì)及其利用進(jìn)行了系統(tǒng)的研究。1.通過對40只出生日期相接近的公羔0~180日齡間的連續(xù)觀測,獲得如下研究結(jié)果:(1)遼寧絨山羊公羔的性行為發(fā)育較早,各個性行為在斷奶后15~30d內(nèi)全部出現(xiàn)并完善。(2)遼寧絨山羊公羔在出生后生長發(fā)育極為迅速,生后一個月齡時的體重為初生時的3倍以上,而斷奶后至6月齡體重增加了近1倍左右。(3)遼寧絨山羊公羔的主要生殖器官均呈現(xiàn)隨日齡增長而迅速發(fā)育的趨勢,6月齡時單側(cè)睪丸、單側(cè)附睪、陰莖長度和精細(xì)管直徑分別為初生時的238倍、53倍、7.2倍和6.14倍,以單側(cè)睪丸的重量生長發(fā)育最為迅速。(4)遼寧絨山羊公羔睪丸內(nèi)的精原細(xì)胞數(shù)量自出生后呈逐漸增加的狀態(tài),30日齡時出現(xiàn)了初級精母細(xì)胞,60日齡時出現(xiàn)次級精母細(xì)胞,90日齡時出現(xiàn)精子細(xì)胞和精子,180日齡時精子細(xì)胞數(shù)量開始減少而精子開始增加;遼寧絨山羊公羔在4月齡可以采集出精液,但精液品質(zhì)中的各個指標(biāo)均低,尚不可進(jìn)行受精,但5月齡之后則其精液品質(zhì)指標(biāo)已經(jīng)與成年公羊相接近。(5)隨著遼寧絨山羊公羔日齡的增長,其外周血清中睪酮的分泌水平也呈逐漸增加狀態(tài),30日齡以后的分泌水平分為為初生時的2.23倍、5倍、7.94倍、10.41倍、13.45倍和16.16倍。說明公羊的性行為決定公羊的種用性能和使用方向與年限;公羊初配與年齡和體重有關(guān);睪丸的重要機能就是產(chǎn)生精子和分泌睪酮。2.通過對遼寧絨山羊8月齡~18月齡、18月齡~24月齡、36月齡以上不同年齡公羊在春夏秋冬4個季節(jié)內(nèi)的精液物理性狀檢測,獲得如下研究結(jié)果:各個年齡段的公羊的采精量和精子活力均是秋季優(yōu)于春季、冬季,春季和冬季優(yōu)于夏季,幼齡公羊的精子活力優(yōu)于成年公羊;不同年齡階段的精子密度均隨季節(jié)發(fā)生變化,精子密度以秋季為最高;遼寧絨山羊成年公羊的頂體完整率以秋季為最高,而精子畸形率為最低,夏季公羊的頂體完整率最低而精子畸形率卻最高;經(jīng)典的卵黃稀釋液稀釋和無動物源性稀釋液稀釋法一樣,在低溫保存狀態(tài)下,精液稀釋倍數(shù)越大,精子體外存活的時間越長;溫度越高,兩種稀釋方法在同一倍數(shù)下,精子的體外存活時間在逐漸減少。經(jīng)典的卵黃稀釋液稀釋法,在同一溫度下,精子在體外的存活時間,隨著稀釋倍數(shù)的增加而逐漸延長;但無動物源性稀釋液稀釋法,在同一溫度下,精子在體外的存活時間,則隨著稀釋倍數(shù)的增加而逐漸減少。說明,幼齡公羊的精子活力優(yōu)于成年羊,應(yīng)選取幼齡公羊作為種公羊且最佳配種季節(jié)為秋季,可獲得較高的受胎率。3.采用熒光定量PCR法對2月齡和15月齡遼寧絨山羊公羊睪丸的Bax和Bcl-2基因的mRNA表達(dá)的差異進(jìn)行研究,獲得如下研究結(jié)果:在遼寧絨山羊睪丸性成熟前后,Bax基因和Bcl-2基因的mRNA均有表達(dá),Bax基因mRNA的表達(dá)量差異極顯著(P0.01),Bax基因性成熟后的表達(dá)量是性成熟前的120倍,Bcl-2基因的mRNA的表達(dá)量差異不顯著(P0.05),Bcl-2基因性成熟后的表達(dá)量是性成熟前的1.4倍,說明性成熟前后均有細(xì)胞凋亡現(xiàn)象,但以性成熟后為主。
[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.
【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S827

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