和田羊与策勒黑羊血液中Mel变化规律和卵巢子宫MTRN1A的表达研究

发布时间：2018-03-04 14:22

本文选题：策勒黑羊　切入点：和田羊　出处：《塔里木大学》2017年硕士论文　论文类型：学位论文

【摘要】：新疆和田地区的策勒黑羊与和田羊是当地主要羊种,前者具有性成熟早、全年发情和密集产羔的特点,后者为半粗毛羊,羊毛品质好,但繁殖率低,为季节性发情动物。有研究报道,褪黑激素(melatonin,Mel)在一定程度上能对动物繁殖性状产生影响,本试验旨在通过对不同品种绵羊褪黑激素与褪黑激素受体表达的时空互作关系的研究,探讨Mel对和田羊与策勒黑羊繁育机能的影响。本次试验采集年周期下春分日、夏至日、秋分日和冬至日四个节气当日早晨、中午、傍晚和凌晨四个时间点的血液,并通过ELISA法测定每个时间点血液中Mel浓度,同时采集和田羊与策勒黑羊的卵巢和子宫组织并提取RNA,采用半定量RT-PCR技术和免疫组化方法,对年周期下和田羊与策勒黑羊Mel分泌、褪黑激素受体(MTRN1A)基因在mRNA水平与蛋白水平的表达情况进行研究。结果表明:日周期下,策勒黑羊血液中Mel浓度在春分当日中午最低,显著低于早晨、傍晚和凌晨(P0.05),傍晚最高,但差异不显著(P0.05);和田羊最高值与最低值出现时间同策勒黑羊,但四个时间点两两之间的差异并不显著(P0.05)。夏至日两种绵羊均是傍晚最高、早晨最低,但差异并不显著(P0.05);秋分日策勒黑羊凌晨浓度显著高于其余时间(P0.05),早晨、中午差异不显著(P0.05),但二者显著低于傍晚(P0.05);和田羊总体变化与策勒黑羊相同,但四个时间点之间并无显著差异(P0.05)。冬至日两种绵羊傍晚含量显著高于其它时间点(P0.05),早晨、凌晨之间差异不显著(P0.05),但均显著高于中午(P0.05)。年周期下,策勒黑羊春分Mel浓度显著高于夏至与秋分(P0.05),秋分显著低于冬至(P0.05),夏至显著低于秋分、冬至(P0.05);同时,春分日卵巢中MTRN1A在mRNA和蛋白水平上,表达量均最高,夏至则最低,显著低于冬至和秋分(P0.05),同时显著低于春分(P0.05),受体表达量则是秋分最低,显著低于冬至(P0.05),但与夏至日差异不显著(P0.05)。子宫中MTRN1A在基因与蛋白均在夏至日表达量最低,且与秋分日并无显著差异(P0.05),但显著低于春分(P0.05),三个季节同时显著低于冬至(P0.05)蛋白表达量。而和田羊则是血液Mel含量冬至显著高于夏至与秋分(P0.05),与春风差异并不显著(P0.05),夏至最低,与秋分差异并不显著(P0.05),但二者显著低于春分(P0.05);卵巢中MTRN1A基因表达量在冬至最高,显著高于春分与夏至(P0.05),与秋分差异并不显著(P0.05),夏至日最低,显著低于春分(P0.05)MTRN1A蛋白的表达量夏至最低,与秋分差异不显著(P0.05),但二者显著低于春分(P0.05),同时,三者显著低于冬至(P0.05);和田羊子宫内MTRN1A的基因与受体表达规律均与策勒黑羊相同。研究表明,两种绵羊血液中Mel含量均存在明显的昼夜节律和季节性节律,但策勒黑羊血液中Mel总体含量高于和田羊,且昼夜变化较大;同样,两种绵羊卵巢和子宫组织中MTRN1A基因与受体的表达均呈现先降后升的季节性变化规律,春分日策勒黑羊卵巢中基因表达量显著高于和田羊,秋分则相反,其他季节无显著差异;同时子宫中的mRNA表达也无显著不同。同样,策勒黑羊春分日卵巢中MTRN1A受体的表达显著高于和田羊,冬至日则相反,其余季节并无显著差异;春分日和田羊子宫中受体的表达显著高于策勒黑羊,其余季节则无显著差异。说明年周期下血液中Mel浓度、MTRN1A基因与蛋白表达量的节律性变化规律,因品种、组织不同而异。本次试验通过对策勒黑羊与和田羊血液中Mel含量变化、子宫卵巢中MTRN1A在m RNA水平和蛋白水平表达的动态规律研究,旨在为研究Mel对不同品种绵羊繁育机能的影响提供一定参考,对实际生产中,规模化养殖场的饲养管理提供一定的科学依据。
[Abstract]:Xinjiang Hetian area of Qira black sheep and Hetian Sheep are the main local sheep, the former has early sexual maturity, characteristics of annual estrus and intensive lambing, the latter half coarse wool sheep, wool quality is good, but the low reproductive rate, seasonal oestrus. Animal studies have reported that melatonin (melatonin, Mel) can have an impact on animal reproductive traits to a certain extent, this experiment aims to study the relationship between the interaction of melatonin and melatonin receptor expression in temporal sheep of different varieties, the effect of Mel on the Hotan sheep and Qira black goat breeding function. The influence of the test collection cycle years the spring equinox, summer solstice, four solar term the autumnal equinox and winter solstice day morning, noon, evening and at four time points of the blood, and through the ELISA method for the determination of Mel concentration in each time point in the blood, while collecting the Hetian Sheep and Qira black goat ovaries and uterus tissue and extract RNA, By using semi quantitative RT-PCR technique and immunohistochemical method, the annual cycle under the Hotan sheep and Qira black sheep Mel secretion of melatonin receptor (MTRN1A) gene expression in mRNA level and protein level. The results showed that the diurnal cycle of Qira black sheep, in the blood concentration of Mel in the first day of spring was lowest at noon, below the morning, evening and morning (P0.05), the highest in the evening, but the difference was not significant (P0.05); and the highest value of the same time Hetian Sheep of Qira black sheep and the lowest value, but no significant difference between the four time points of 22 (P0.05). The summer solstice two sheep were lowest in the morning and evening is the highest. But the difference was not significant (P0.05); Qira black sheep at the equinox were significantly higher than the rest of the time (P0.05) morning, noon, the difference was not significant (P0.05), but the two was significantly lower than that in the evening (P0.05); Hetian Sheep and Qira black sheep of the same overall change, but the four time points There is no significant difference between (P0.05). The winter solstice two evening of sheep was significantly higher than that of other time points (P0.05), there was no significant difference between the early morning (P0.05), but significantly higher than at noon (P0.05). The annual cycle, Qira black sheep Mel was significantly higher than the spring equinox and the autumnal equinox to the summer (P0.05). The autumnal equinox was significantly lower than that of winter solstice (P0.05), was significantly lower than that of summer solstice equinox, winter solstice (P0.05); at the same time, the spring equinox in the ovary of MTRN1A in mRNA and protein level, the expression was the highest, the summer solstice is the lowest, significantly lower than that of the winter solstice and equinox (P0.05), and was significantly lower than that of the vernal equinox (P0.05), receptor expression is the lowest was significantly lower than that of the winter solstice (P0.05), but the difference between the summer solstice (P0.05). MTRN1A gene and protein in the uterus were the lowest expression in the summer solstice and equinox, and there was no significant difference (P0.05), but significantly lower than that in spring (P0.05), the three seasons and was significantly lower than that of the winter solstice (P 0.05) protein expression. While the Hetian Sheep is the blood content of Mel was significantly higher than that of the winter solstice summer solstice and equinox (P0.05), the spring breeze and the difference was not significant (P0.05), the lowest, and the difference was not significant (P0.05), but the two was significantly lower than that of the spring equinox (P0.05); ovarian expression of MTRN1A gene in the winter solstice the highest, significantly higher than the spring equinox and solstice (P0.05), and the difference was not significant (P0.05), the lowest was significantly lower than that of the summer solstice, the vernal equinox (P0.05) the expression of MTRN1A protein was the lowest and the summer solstice, the difference was not significant (P0.05), but the two was significantly lower than that of the spring equinox (P0.05), at the same time, the three was significantly lower than that of the winter solstice (P0.05); and the receptor gene and the expression of MTRN1A in uterine Hetian Sheep rules are the same with the Qira black sheep. The study shows that the Mel contents of two sheep blood existed in the circadian rhythm and seasonal rhythm is obvious, but the Qira black sheep blood total Mel was higher than that in Hetian Sheep, and day and night Large changes; similarly, the expression of MTRN1A two gene in sheep ovarian and uterine tissue and receptor showed seasonal variation rose after the first drop, the gene expression of the equinox of Qira black sheep ovarian volume was significantly higher than that of Hetian Sheep, the contrary, no significant difference in other seasons; at the same time, the expression of mRNA in uterus no significant different. Similarly, the expression of MTRN1A receptor of Qira black sheep ovary was significantly higher than that of the spring equinox and winter solstice Tian sheep, on the contrary, there is no significant difference in other seasons; spring equinox and receptor expression in the uterus of Hetian Sheep was significantly higher than that of Qira black sheep, there was no significant difference in other seasons. The blood concentration of Mel in the year cycle next, MTRN1A gene and protein expression rhythmicity, the amount due to variety, different organizations. Through this experiment of Qira black sheep and Hetian Sheep Blood Mel content change, ovary and uterus in M RNA and MTRN1A The study of the dynamic regulation of protein level aims to provide some references for the study of the effects of Mel on the breeding performance of different breeds of sheep, and provide a scientific basis for the breeding and management of large scale farms in actual production.

【学位授予单位】：塔里木大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S826

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