【作者】 傅启高；

【导师】 陈杰；

【作者基本信息】 南京农业大学 ， 基础兽医学， 2000， 博士

【摘要】 本文在测定了绍兴鸭和高邮鸭产蛋性能的基础上，比较研究了绍兴鸭和高邮鸭卵巢卵泡生长发育的变化规律及其与产蛋性能的关系，探讨了绍兴鸭优良产蛋性能的内分泌及分子生物学机制。一，绍兴鸭和高邮鸭产蛋基本规律的比较研究 取14周龄圈养绍兴鸭和高邮鸭各102只和52只，记录鸭开产后每天的产蛋率和蛋重，及环境因素的变化。结果表明，绍兴鸭比高邮鸭开产早（1周龄），50％以上产蛋率维持时间长（39vsl6周）。突然变换日粮、环境温度、饲养密度和换羽明显影响产蛋性能。 取14周龄笼养绍兴鸭85只，单只笼养，填写个体产蛋记录，分析产蛋基本规律。结果表明，绍兴鸭有2个主产蛋序列，连续产蛋天数分别为31.62±2.94和36.33±2.34天；连续停产天数分别为1.91±0.27和4.55±0.68天。 与圈养鸭相比，在产蛋中后期（24～59周龄），笼养绍兴鸭平均产蛋率（85.99±1.25％ vs 78.67±1.91％，p＜0.002）和产蛋后期（51～59周龄）平均蛋重（72.63±0.54克 vs 70.87±0.30克，p＜0.01）都较高。整个产蛋期的产蛋率（77.74±3.21％ vs 73.52±2.75％，p＜0.05）和蛋重（66.87±1.3克 vs66.35±1.12克，p＞0.05）也都较高。二，绍兴鸭和高邮鸭卵巢卵泡发育基本规律的比较研究 在14、23、28、36、40、51和59周龄随机取圈养绍兴鸭和高邮鸭以及笼养绍兴鸭各8只屠宰，观测卵巢发育情况，记录卵巢重和排卵前卵泡数。结果表明，绍兴鸭卵巢发育成熟早于高邮鸭（17周龄vs37周龄），圈养绍兴鸭卵巢重显著低于高邮鸭，绍兴鸭卵泡闭锁少见而高邮鸭则较常见。 圈养绍兴鸭卵巢重和大卵泡数与蛋重呈显著正相关，圈养高邮鸭卵巢重和大卵泡数与产蛋率和蛋重均呈显著负相关；笼养绍兴鸭卵巢重和大卵泡数与产蛋率呈显著正相关。 取59周龄圈养绍兴鸭和高邮鸭各5只屠宰，测定第1（F₁），第3（F₃），第5（F₅）大卵泡和直径小于8mm的小卵泡（SF）细胞的增殖指数和凋亡率。结果表明，绍兴鸭SF细胞早期和累积凋亡率显著高于高邮鸭，细胞增殖指数显著低于高邮鸭。绍兴鸭和高邮鸭SF细胞的累积凋亡率和增殖指数显著高于F₅～F₁细胞。 傅启高；绍兴瞩优良产蛋性愧的内分泌及分子生物 制研灾三，绍兴瞩和高邮瞩血清生长激素结合蛋白结合活性（GHBP lA）、生长激素（Gff和雌二醇田二冰平的比效研究 比较测定绍兴鸭和高邮鸭血清GHBPSA、OH和EZ水平。结果表明，绍兴鸭血清EZ水平显著高于高邮鸭。笼养和圈养绍兴鸭血清GHBP8A与卵巢重和大卵泡个数负相关。四，绍兴瞩和高邮瞩卵巢卵泡黄体生成运受体0HR人卵泡刺澈索 受体（FSRR）和生长激蠢受体（GHR因表达的比较研究 比较测定绍兴鸭和高邮鸭卵巢中h、F3、FS和 SF卵泡膜上L皿、FS皿和GHR基因的表达水平。结果表明，两种鸭同等级卵泡上LHR、FSHR和GHR基因的表达水平没有明显的差异，随卵泡发育呈一致的趋势。F3和FS卵泡 LHR基因的表达水平高于 SFto＜0．05X而 FS皿和 OHR基因的表达水平低于8＊O＜0．05）。五，绍兴瞩卵巢卵泡颗粒细胞和膜细胞LRR、FSRR和GHR基 因表达的研究及 LHR和 FSHR cDNA序列分析 65周龄绍兴鸭4只，取其FI、F3、F，、以及新进入等级的卵泡（NRP）和小黄卵泡侣），分离膜细胞和颗粒细胞，测定细胞LHR、FSHR和 GHR基因的表达水平。结果表明，卵巢发育备阶段卵泡膜细胞GllR基因的表达水平要比颗粒细胞高，其中FI差异显著O＜0．05卜FI，FS和N’RPF颗粒细胞FSHR基因的表达水平高于膜细胞O＜O．05卜民和8＊F颗粒细胞FS皿基因的表达水平也有高于膜细胞的趋势。nO＜o．05卜＊y F和S WW卜o．01)膜细胞＊＊R基因的表达水平高于颗粒细胞。 卵巢卵泡颗粒细胞 LHR基因的表达水平随着卵泡的成熟而升高O＜0刀 1)，而颗粒细胞和膜细胞GHR和FSHR基因。膜细胞LHR基因的表达则未见有这种规律。 LHR不 FSHR CDNA序列分析的结果表明，百泡 LHR CDNA（膜外结构域部们序列在 89、189和 349位分别有 Sacl、Ekorx和 hOI切点。核苦酸序列与鹤鸦（468b）和鸡O）的同源性分别为88．46％和89．74％，由此椎导的氨基酸序列与鹤鸦156ed和鸡（156eq的同源性分别为po．23％和95．sl％。FSHRcDNA（膜外结构域部分）核昔酸＊2】M和由此推导的氨基酸＊）序列与鸡的同源性分别为 89．96％和 89．47％。绍兴鸭在 2！位（相对于鸡的 328位）和 5 6位湘对于鸡的 823位)碱基不明(或缺失＞绍兴鸭n＊位(鸡的 718位X碱基变换为 c碱基，EcOR切点消失。2 摘 要 六，GH对绍兴鸭卵巢卵泡颗粒细胞和膜细胞LHR、FSHR和 GHR基因表达及孕回和E。分泌的彤响 取40周龄绍兴鸭，无菌获取民，F3，F。，N’－－－F和SYF，分离膜层和颗 粒层，收获细胞，在无血清MI99培养液中培养，并用鸡重组GH处理。结果更多还原

【Abstract】 Endocrinological and molecular biological mechanisms of excellent egg performance in Shaoxing ducks were examined in the present paper, involving following aspects: 1. Comparison of egg performance between Shaoxing and Gaoyou ducks Laying rate and egg weight of 102 Shaoxing ducks and 52 Gaoyou ducks reared on floor were recorded. The results showed that the onset of lay in Shaoxing ducks were earlier than that in Gaoyou ducks (17 vs 37 weeks of age), the duration of 50% laying rate was longer in Shaoxing ducks than in Gaoyou ducks (39 vs 16 weeks). In addition, acute change of diet, molting, temperature and ducks density influenced the egg performance in Sbaoxing ducks. Laying rate and egg weight of 85 Shaoxing ducks reared in cage were recorded. The sequence length and the pause length were analyzed. There were two main sequence lengths that are 31.62+2.94 and 36.33+2.34 days respectively, while pause lengths are 1.91+0.27 and 4.55?.68 respectively. Laying rate of cage reared Sbaoxing ducks was improved compared to that of floor reared ducks (77.74?.21 vs 73.52?.75%, p<0.05), while egg weight (66.87?1.3 vs 66.35?.12 gram) was not affected. 2. Developmental changes of ovarian follicles, follicular cellular proliferation and apoptosis in Shaoxing and Gaoyou ducks during lay Ovary were sampled from 8 Shaoxing and Gaoyou ducks reared in cage versus on floor at 14, 23, 28, 36, 40, 51 and 59 weeks of age respectively. Their ovary weight and numbers of preovulatory follicles were recorded. The results showed that the maturity of ovary was earlier in Shaoxing ducks than that in I0I ~L*4~F~ Gaoyou ducks (17 vs 37 weeks of age). The ovary was heavier in floor reared Gao you ducks than that in Shaoxing ducks. The follicles atresia was common in Gaoyou ducks while seldom in Shaoxing ducks. There are strong positive correlations between ovary weight, preovulatory follicle numbers and laying rate, egg weight in cage versus floor reared Shaoxing ducks, while strong negative correlations in floor reared Gaoyou ducks. The first, the third and the fifth largest preovulatory follicles (F1, F3 and F5), and the small follicles (SF, diameter < 8 mm) were sampled from 5 Shaoxing and Gaoyou ducks reared on floor at 59 weeks of age respectively. Cell proliferation and apoptosis were detected using the f]owcytonietiy. The results showed that the percentages of overall and early cell apoptosis of SF were significantly higher in Shaoxing ducks than in Gaoyou ducks, while the proliferation index were lower in Shaoxing ducks than in Gaoyou ducks. 3. Changes of serum levels of growth hormone(GH), 17- ~ estradiol (E2) and binding activity of GH binding protein (GHBP-BA) in Shaoxing ducks during iay Sera were prepared from the 8 Shaoxing and Gaoyou ducks at different weeks of age respectively above-mentioned, their levels of OH and E2 were analyzed using the radio-immuno-assay (RIA), Gl-IBP-13A was analyzed using the radio -binding assay. The results showed that serum E2 level was higher in Shaoxing ducks than in Gaoyou ducks. There are strong negative correlations between serum level of GHBP-BA and ovary weight and preovulatoiy follicle numbers in Shaoxing ducks reared cage versus floor.更多还原