尼罗罗非鱼雌性性腺关键LncRNA的鉴定和表达分析

生殖细胞的分化与发育是硬骨鱼类性别分化的重要环节,并受到遗传和环境2个因素的影响.为研究LncRNA在硬骨鱼类卵母细胞发育以及性别分化中的作用,课题组通过转录组测序获得了尼罗罗非鱼(Oreochromis niloticus)雌、雄性腺中呈现差异表达的LncRNA,并选取LncRNA(TCONS_02477925)开展研究.发现该LncRNA具有2个外显子,属于典型的基因间LncRNA(Intergenic LncRNA),转录本全长1 537 nt,基因组定位于19号连锁群(Linkage Group 19,LG19)上,并且该基因在卵巢的表达水平显著高于精巢.荧光原位杂交结果表明,该LncRNA主要表达在I和II时相的卵母细胞的细胞质中.转录组测序结果及生物信息学分析发现,qrsl1和rtn4ip1可能是该LncRNA的cis靶基因,并且2个靶基因在卵巢中表达丰度较高.因此,本研究表明该LncRNA在罗非鱼卵母细胞发育和性别分化中可能有重要的作用.     

猪卵母细胞生长成熟过程中脂质成份的研究

本实验应用Ｃａｔａｌａｎ及Ｌｉｌｌｉｅ氏油红Ｏ染色法和Ｂａｋｅｒ氏酸性氧化苏木精染色法，对猪卵母细胞生长成熟过程中脂质成份的改变情况进行了染色研究，把生长卵泡按其直径：０．８～２．４ｍｍ（Ｓ组）、２．５～４ｍｍ（Ｍ组）、＞４ｍｍ（Ｌ组）分为三组，分别取其中的卵母细胞进行染色观察．另外，还作了卵巢冰冻切片的染色观察．结果表明，在卵泡腔形成之前的猪卵母细胞中就含有大量的脂滴，在卵泡形成时，卵母细胞内的脂滴有所增加．脂滴中含有中性脂肪和磷脂．     

长毛对虾卵母细胞环形片层的研究

应用电镜技术研究长毛对虾(Penaeus penicillatus)卵母细胞中的环形片层(AL)。AL在卵黄发生前的卵母细胞卵质中出现,在卵黄发生期间消失。AL由8条平行排列,具有50～70 nm 环孔的片层组成。相邻片层之间的距离约为70 nm。AL 通常没有核糖体附在膜片层的外表面。AL末端膨大,与高尔基体毗邻。从形态学上看,AL可能来源于高尔基体;高尔基膜囊伸直、拉平,并局部融合形成环孔,最后演变成AL。卵黄发生期间,AL分化形成为许多囊泡,为卵黄粒的形成提供沉积的场所。本文还讨论高尔基体、AL 及卵黄粒形成之间的关系。     

Effect of leptin on oocyte maturation and subsequent pregnancy rate of cloned embryos reconstructed by somatic cell nuclear transfer in pigs

Cloning pigs by somatic cell nuclear transfer （SCNT） has wide applications in basic research, human medicine and agricultural production. To improve cloning efficiency, the effect of two basic maturation media, NCSU-23 and TCM199, was compared, and TCM199 was selected for the following experiments with leptin. We systematically studied the effects of leptin supplementation on oocytes in vitro maturation （IVM）, in vitro development of parthenogenetically activated （PA） and SCNT embryos and in vivo development of SCNT embryos after embryo transfer （ET）. The results showed that supplementation of 100 or 200 ng/ml leptin into the maturation medium did not greatly affect nuclear maturation of oocytes, or cleavage rates of PA and SCNT （P 〉 0.05）. Blastocyst rates of PA and SCNT embryos were significantly improved when 100 or 200 ng/ml leptin was added to maturation medium, and the number of cells in PA blastocysts was also improved （P 〈 0.05）. The number of cells in blastocyst of SCNT was improved, when 100 ng/ml leptin was added （P 〈 0.05）. Furthermore, supplementation of 100 or 200 ng/ml leptin to the IVM medium may improve pregnancy rate and the delivery rate inpig cloning.