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.     

KK-42对家蚕蛹血淋巴及卵巢蛋白质合成的影响

研究了咪唑类化合物KK—42对家蚕卵巢发育以及蛋白质合成的影响,结果表明KK-42对卵巢增重有明显的抑制作用,血淋巴蛋白含量在蛹期变化幅度很小,卵巢蛋白质含量增加也很缓慢;而对照区在化蛹后,血淋巴蛋白急剧增加,第三天达到高峰,以后又迅速下降,与卵巢蛋白质增加相吻合。由此表明,咪唑类化合物KK-42不仅抑制血淋巴蛋质的合成,同时还抑制了血淋巴蛋白向卵巢的转移。     

Interaction between melatonin and follicle-stimulating hormone promotes in vitro development of caprine preantral follicles

The aim of this study was to investigate the effects of melatonin and follicle-stimulating hormone (FSH) on the in vitro culture of goat preantral follicles. Ovarian fragments were cultured for 7 d in α-minimum essential medium (α-MEM⁺) containing melatonin (100, 250, 500, or 1,000 pM), FSH (50 ng/mL), or a combination of the 2 hormones and further analyzed by histology and transmission electron and fluorescent microscopy. The results showed that after 7 d of culture, tissues cultured in α-MEM⁺ alone or supplemented with FSH alone, melatonin (500 and 1,000 pM), or the combination of FSH and melatonin (1,000 pM) maintained percentages of normal preantral follicles similar to the fresh control. In contrast to the noncultured tissues, the percentage of developing follicles was increased under all culture conditions after 7 d (P < 0.05). The addition of 1,000 pM melatonin associated with FSH to the culture medium increased follicular and oocyte diameters compared with α-MEM⁺ alone after 7 d of culture (P < 0.05). Ultrastructural and fluorescent analyses confirmed the integrity of follicles cultured with 1,000 pM of melatonin plus FSH for 7 d. In conclusion, this study demonstrated that the interaction between melatonin and FSH maintains ultrastructural integrity and stimulates further growth of cultured caprine preantral follicles.     

Expression Pattern of Vascular Endothelial Growth Factor in Canine Folliculogenesis and its Effect on the Growth and Development of Follicles after Ovarian Organ Culture

In this study, the expressions of VEGF in dog follicles were detected by immunohistochemistry and the effects of VEGF treatment on the primordial to primary follicle transition and on subsequent follicle progression were examined using a dog ovary organ culture system. The frozen‐thawed canine ovarian follicles within slices of ovarian cortical tissue were cultured for 7 and 14 days in presence or absence of VEGF. After culture, the ovaries were fixed, sectioned, stained and counted for morphologic analysis. The results showed that VEGF was expressed in the theca cells of antral follicles and in the granulosa cells nearest the oocyte in preantral follicle but not in granulosa cells of primordial and primary follicles; however, the VEGF protein was expressed in CL. After in vitro culture, VEGF caused a decrease in the number of primordial follicles and concomitant increase in the number of primary follicles that showed growth initiation and reached the secondary and preantral stages of development after 7 and 14 days. Follicular viability was also improved in the presence of VEGF after 7 and 14 days in culture. In conclusion, treatment with VEGF was found to promote the activation of primordial follicle development that could provide an alternative approach to stimulate early follicle development in dogs.     

玉米未授粉子房离体培养及植株再生

探讨了不同材料、不同培养基对玉米未授粉子房的影响。确定了未授粉子房最佳培养时期,筛选出一批用于玉米未授粉子房诱导频率高的培养材料。     

中国对虾卵黄蛋白原合成部位的研究

十足目甲壳类卵巢中卵黄的来源,在过去的几十年研究中一直存在争议,内源性合成和外源性合成都有报道。本文以雌性中国对虾为实验材料,根据组织学观察确定其发育阶段,试验虾可以分为:卵巢未发育期;卵黄发生前期;初级卵黄发生期和次级卵黄发生期4个时期。从处于不同卵巢发育期对虾的卵巢和肝胰腺中提取总RNA,用RT-PCR的方法初步探讨不同发育期的中国对虾卵巢和肝胰腺中卵黄蛋白原mRNA的表达,确定是否存在卵黄蛋白原的合成功能。结果发现,从卵巢未发育期到次级卵黄发生期的卵巢和肝胰腺中都有卵黄蛋白原mRNA的表达,说明二者为卵黄蛋白的合成部位。     

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。     

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。     

鸡卵巢传入神经元在脊神经节内的节段性分布

将CB—HRP(霍乱毒素B亚单位结合态的辣根过氧化物酶)溶液注入母鸡卵巢内,逆行追踪研究了分布于卵巢的感觉神经元的定位及其规律性。结果表明:①分布于鸡卵巢,经交感途径传入的感觉神经元胞体位于T_1—L_3节段的脊神经节内,峰值在T_6节段。说明鸡卵巢经交感途径传入的感觉神经元的分布具有相对的集中性和弥散性。②脊神经节内的标记细胞以小细胞居多,推测鸡卵巢内可能含感受损伤性刺激的纤维占绝对优势。