牛精子不同获能方法对体外受精效果的影响

为了提高牛体外受精率,探讨牛冷冻精液不同获能方法对体外受精效果的影响,对牛冷冻解冻精液进行了两次离心获能法与一次离心加上游法获能对体外受精效果影响试验.结果表明,两次离心组受精率卵裂率(75.6-±4.5)％与一次离心加上游法组受精卵裂率(76.4±1.9)％无显著差异(P＞0.05);二次离心组囊胚率(35.7±4.1)％与一次离心加上游法组(36.3±2.7)％差异不显著(P＞0.05).而两次离心组却可以大大节省了体外获能时间.     

白血病抑制因子（LIF）及其受体在月经 周期猕猴卵巢内的表达

应用免疫细胞化学方法对白血病抑制因子（leukemia inhibitory factor, LIF）、白血病抑制因子受体（leukemia inhibitory factor receptor, LIFR）和gp130在月经周期猕猴卵巢内的表达进行了研究。结果表明：LIF、LIFR和gp130在猕猴卵巢内的表达量随月经周期的不同而变化，LIF及其受体在增殖期卵巢内的表达量高于分泌期的卵巢。LIF及其受体在原始卵泡、腔前卵泡和有腔卵泡的卵细胞内表达量均较高，在颗粒细胞、卵泡膜、卵巢生殖上皮和卵巢基质中有少量表达，而在闭锁卵泡、退化的黄体和卵巢髓质中未见表达。LIF及其受体在猕猴卵巢内的表达可能受卵巢分泌的类固醇激素所调控，LIF可能通过自分泌或旁分泌的方式在猕猴卵泡发育及排卵等过程中起重要作用。     

山羊转t-PA突变体基因的体细胞核移植研究

比较了用转t-PA突变体基因的胎儿成纤维细胞和卵丘细胞作供体时核移胚的发育情况。结果表明：卵丘细胞作供体时，核移胚的卵裂率（82.9%）和桑葚胚率（55.9%）均高于胎儿成纤维细胞卵裂率（75.7%）和桑葚胚率（48.7%），但差异不显著（P＞0.05）；卵丘细胞作供体时的囊胚率（19.7%）则高于胎儿成纤维细胞（11.0%），差异显著。(P<0.05)。比较了饥饿处理与否对胎儿成纤维细胞作供体时转基因核移胚的发育情况。结果表明：供体细胞饥饿后，其核移胚的卵裂率（75.6%）与供体不饥饿的卵裂率（75.8%）差异不显著；供体细胞饥饿后，其核移胚的桑葚胚率（46.5%）和囊胚率（11.0%）高于供体不饥饿的桑葚胚率（39.3%）和囊胚率（9.0%），但差异不显著。比较了SOFaa和CR1aa分别添加10%FBS和bFF后，转基因核移胚的发育情况。结果表明：用CR1aa液培养时，添加10%bFF，其囊胚率（9.1%）高于添加10%FBS（4.2%）（P＞0.05）；用SOF液培养时，添加10%bFF，其囊胚率（11.2%）高于添加10%FBS（5.5%）（P＞0.05）。说明用SOF液优于CR1aa液。     

Role of ghrelin in regulating rabbit ovarian function and the response to LH and IGF-I

The aim of these in vivo and in vitro studies was to examine the role of ghrelin in the control of plasma hormone concentrations, the proliferation, apoptosis and secretory activity of ovarian granulosa cells and the response of these cells to hormonal treatments. Female rabbits were injected with ghrelin (10 μg/animal/day for one week before ovulation induced by 25 IU PMSG and 0.25 IU LHRH). On the day of ovulation, blood samples were collected and analyzed for concentrations of progesterone (P₄), testosterone (T), estradiol (E₂), estrone-sulphate (ES), insulin-like growth factor I (IGF-I) and leptin (L) by RIA. Some control and ghrelin-treated animals were killed in the periovulatory period, their ovaries were weighed and granulosa cells were isolated and cultured for 2 d. Cell proliferation (expression of PCNA) and apoptosis (expression of TdT) were evaluated by immunocytochemistry and TUNEL respectively. Secretion of P₄, T, E₂, IGF-I, and prostaglandin F (PGF) by granulosa cells cultured with and without LH or IGF-I (1, 10 or 100 ng/ml medium) was assessed by RIA. The remaining control and treated animals were kept until parturition, while the number, viability and body weight of pups were recorded.     

Histomorphological and immunohistochemical study of angiogenesis and angiogenic factors in the ovary of the mare

Cyclical ovaries of 18 mares were examined histologically and immunohistochemically for vascular endothelial growth factor A and B (VEGF A; VEGF B), angiopoietin1 and 2 (Ang1; Ang2), vascular endothelial growth factor receptor 1 and 2 (VEGF-R1; VEGF-R2), angiopoietin receptor (Tie2) and von Willebrand factor. The most intensive coexpression of the examined factors and receptors was detected in the periovulatory period, when a distinctive ovarian angiogenesis takes place, being essential for tertiary follicle maturation and for the endocrine function of the Corpus luteum. Based on the immunohistochemical results, VEGF A, Ang2, VEGF-R2 and Tie2 in particular seem to play a significant role on angiogenesis during follicular and luteal development in the mare, while Ang1 supports vessel stabilisation. The findings of luteal regression and follicular atresia showed that, in the absence of VEGF A, Ang2 and its receptor Tie2 contribute substantially to vessel regression and therefore to luteolysis and follicular atresia.     

Multiple molecular forms of inhibin in buffalo (Bubalus bubalis) ovarian follicular fluid

Inhibin is a heterodimeric glycoprotein hormone involved in the regulation of FSH release from the anterior pituitary gland and it has been characterized from various animals. Although, multiple molecular forms of inhibin have been reported from different species, however, the molecular nature of inhibin has not been studied in buffaloes. In the present study, attempts were made to identify inhibin in buffalo ovarian follicular fluid. Buffalo ovaries were obtained from the local abattoir and follicular fluid was aspirated from surface follicle (with diameter ?5 mm). A combination of techniques (viz., gel filtration, SDS-PAGE, Western blot etc.) was employed for identification and isolation of inhibin(s). Inhibin bands were detected at 129 and 63 kDa by Western blot analysis in non-reducing conditions. In reduced SDS-PAGE, 63 kDa fraction produced a single band while 129 kDa fraction resolved into four components of 63, 43, 29 and 20 kDa. Out of them only 29, 63 and the native 129 kDa fractions produced bands on Western blot analysis. In total five fractions (63, 54, 39, 29, 25 kDa) were obtained by trypsin digestion of 129 kDa form. However, only 63 and 29 kDa fractions showed immunoreactivity. In this study, for the first time, we have identified two major forms of inhibin (129 and 63 kDa) with little proteolytic cleavage/processing of the large precursor in the buffalo follicular fluid.     

油菜高效转化系统的研究

以甘蓝型油菜湘油13为试验材料，运用子房注射法将BT毒蛋白基因导入油菜。在授粉后第20小时至第30小时用自制的微玻针从子房中部注射0.5~1.5μg外源DNA，以高达12.8%的频率获得抗卡那霉素植株。分子杂交分析证明BT毒蛋白基因已整合到油菜基因组中，转化频率为1.6%，表明子房注射法是一种有效、实用的油菜遗传转化方法。     

Kiss-1基因在猪初情期下丘脑-垂体-卵巢轴中的定位研究

[目的]为阐明猪的生殖机理和选育早熟品种奠定基础。[方法]采用免疫组化技术研究Kiss-1 mRNA在初生以及30、60、70(初情期)和90日龄小梅山猪的下丘脑、卵巢和垂体内的定位,并利用放射免疫分析(RIA)方法观察母猪血浆LH、FSH含量的变化。[结果]Kiss-1 mRNA在小梅山猪下丘脑、卵巢、垂体中均有表达,以下丘脑弓状核阳性颗粒最多,尤其初情期时最为明显。各级卵泡中以初情期成熟卵泡的阳性颗粒数目最多。Kis-s1 mRNA表达量在小梅山猪下丘脑、垂体和卵巢从初生到初情期逐渐上升,并在初情期达到顶峰,而后逐渐下降。小梅山猪血浆LH、FSH浓度初生时较高,随后迅速下降,60日龄又升高,70日龄达到峰值。[结论]Kiss-1基因是控制猪初情期启动的关键因子     

芥菜型油菜和黑芥与诸葛菜属间杂种的获得及其特性

属间远缘杂交是创造新种质、解决育种材料短缺的一种有效方法。为了使诸葛菜的优良基因转入芥菜型油菜和黑芥中, 采用子房培养技术, 进行芥菜型油菜和黑芥与诸葛菜的正反属间杂交。在6种不同的培养基中培养子房。结果表明, 不同的杂交组合、培养基和母本的属间杂交可交配性不同。通过子房培养, 获得了10株属间杂种, 其中9株在形态上近似母本, 或介于父母本之间, 尤嫩斯与诸葛菜杂交组合的1株杂种表现出完全的雄性不育, 并在分枝类型和叶型方面表现出明显的变异; 形态学和SSR分子生物学鉴定表明10株杂种都是真正的属间杂种。     

克氏原螯虾大颚器促性腺发育作用研究

[目的]研究克氏原螯虾（Procamharus clarkia）大颚器的组织学特点，探讨其大颚器在性腺发育过程的作用。[方法]采用大颚器和性腺组织的离体联合培养法，用卵黄发生期的克氏原螯虾大颚器刺激不同发育期的克氏原螯虾性腺。[结果]2处理组卵巢的最大卵径为（0．32±0．05）mm和（0．27±0．06）mm，2对照组卵巢的最大卵径为（0．24±0．05）mm和（0．20±0．05）mm；2处理组精巢的精母细胞直径为（13．09±0．75）μm和（12．93±0．98）μm，2对照组精巢的精母细胞直径为（10．29±1．00）μm和（10．02±0．68）μm。[结论]数据显示处理组的性腺细胞均比对照纽的性腺细胞显著增大（P〈0．05），表明大颚器的功能性物质具有促进性腺细胞发育的作用，支持大颚器作为促性腺生长发育腺体的观点。