Using Toad Oocytes as an Ultrasensitive System for Estimating the Biological Activity of Trichosanthin

A sensitive bioassay for estimating the activity of Trichosanthin was developed by using intraoocyte microinjection as the technique and germinal vescicle breakdown as a marker. The GVBD inhibitory activity assay was million-fold more sensitive than the routinely used abortifacient activity assay. The detectable amount of Trichosanthin ranged from 0.5 to 100 pg. Since the least detectable dose was 0.5 pg, it made the assay suitable to analyse the microquantity of pure Trichosanthin used for studying the structure and function of the drug.     

A FACTOR FROM GRANULOSA CELLS CAN STIMULATE OOCYTE TISSUE PLASMINOGEN ACTIVATOR ACTIVITY

Several studies indicate that substances synthesized by granulosa cells are capableof regulating oocyte activity. We have studied the effect of factors synthesized by gran-ulosa cells on tPA activity of denuded oocytes using a co- culture system. The resultsshow that an FSH- dependent factor(s) synthesized by granulosa cells (but not by theca-interstitial cells) is capable of stimulating tPA activity of denuded oocytes. This findingis important for understanding hormonal regulation of oocyte tPA activity by mediatorssynthesized in granulosa cells.     

应用共聚焦显微镜观察卵母细胞成熟过程中Cdc42活性变化

Cdc42属于GTP酶家族成员,参与细胞骨架调节和细胞发育,但在细胞分裂过程中的作用所知甚少.以爪蟾卵母细胞为模型,GFP-wGBD(Cdc42活性探针)和罗丹明-微管蛋白作为荧光标记物,采用共聚焦显微镜观察卵母细胞分裂过程中Cdc42活性变化.结果显示卵母细胞中,极体释放数分钟前,纺锤体上方和周边可见微弱的Cdc42活性,2 min～4 min内,活性逐渐增强,随后收缩环形成,胞质分裂发生.胞质分裂完成后,Cdc42活性消失.结果提示Cdc42活性可能与爪蟾卵母细胞胞质分裂有关.     

基于微流控的人卵母细胞特性检测

人的卵母细胞特性的研究对于临床医学具有重要意义,可以通过卵母细胞的大小对其活性进行研究。采用微流控平台与MATLAB算法相结合的方法,利用微流控器件的沟道对卵母细胞进行挤压,由于卵母细胞的活性不同对于挤压的反应也不同,活性越好的卵母细胞通过沟道时变形性越好,经检测卵母细胞有很好的活性;拍摄卵母细胞发生形变与卵母细胞通过沟道的视频,利用MATLAB程序进行图像处理得到卵母细胞的大小,测得卵母细胞的直径约为170μm。这种方法有别于传统方法,在一定程度上使用了算法识别,提高了卵母细胞分析统计的准确度,提升了工作效率。由于人的卵母细胞比较珍贵,只进行了初期实验研究,证明此方法确实可行。     

Osmotic and aging effects in caviar oocytes throughout water and lipid changes assessed by 1H NMR T1 and T2 relaxation and MRI

By combining NMR relaxation spectroscopy and magnetic resonance imaging techniques, unsalted (us) and salted (s) caviar (Acipenser transmontanus) oocytes were characterized over a storage period of up to 90 days. The aging and the salting effects on the two major cell constituents, water and lipids, were separately assessed. T1 and T2 decays were interpreted by assuming a two-site exchange model. At Day 0, two water compartments that were not in fast exchange were identified by the T1 relaxation measurements on the us oocytes. In the s samples, T1 decay was monoexponential. During the time of storage, an increment of the free water amount was found for the us oocytes, ascribed to an increased metabolism. T1 and T2 of the s oocytes shortened as a consequence of the osmotic stress produced by salting. Selective images showed the presence of water endowed with different regional mobility that severely changed during the storage. Lipid T1 relaxation decays collected on us and s samples were found to be biexponential, and the T1 values lengthened during storage. In us and s oocytes, the increased lipid mobility with the storage was ascribed to lipolysis. Selective images of us samples showed lipids that were confined to the cytoplasm for up to 60 days of storage.     

A Review on Microfluidics: An Aid to Assisted Reproductive Technology

Infertility is a state of the male or female reproductive system that is defined as the failure to achieve pregnancy even after 12 or more months of regular unprotected sexual intercourse. Assisted reproductive technology (ART) plays a crucial role in addressing infertility. Various ART are now available for infertile couples. Fertilization in vitro (IVF), intracytoplasmic sperm injection (ICSI) and intrauterine insemination (IUI) are the most common techniques in this regard. Various microfluidic technologies can incorporate various ART procedures such as embryo and gamete (sperm and oocyte) analysis, sorting, manipulation, culture and monitoring. Hence, this review intends to summarize the current knowledge about the application of this approach towards cell biology to enhance ART.     

Intraovarian control of selective follicular growth and induction of oocyte maturation in mammals

In newborn mammals, most of the germ cell population rests in a pool of quiescent small follicles in the ovaries. Regularly throughout adulthood, a small percentage of these oocytes and follicles grows to a certain stage of development and then either degenerates or matures and ovulates. This entire process is under both exogenous and endogenous control. Recent work, including my laboratory’s, has clarified that cytokines and glycosaminoglycans are involved as exogenous and endogenous factors in ovarian follicular development, atresia, and maturation in mammals. The present article describes our contribution regarding the cytokines and ovarian glycosaminoglycans that act as intraovarian regulators of follicular development and oogenesis, including oocyte maturation, in mammals.     

大鲵胚胎发育的组织学研究

通过人工繁殖，获得了两批大鲵的受精卵及其幼苗．利用组织学方法研究了大鲵的早期胚胎发育，并描述了其主要特征．同时，就此和其他两栖动物的胚胎发育进行了比较讨论，并指出了其共性与特殊性．     

非洲爪蟾卵母细胞内源性ATP激活电流ID2成分的分析

在带有滤泡膜的非洲爪蟾卵母细胞标本上,应用双电极电压钳及胞内透析方法进行记录,外加ATP所激活的内源性膜电源包括3个成分依次为：快内向电流（early fast depolarizing current,ID1),慢内向电流（lateslow depolarizing current,ID2)和慢外向电流(slow hygerpolarrizing current,IH),本文主要分析ID2成分的特征及其产生的机制,结果如下：（1）胞内透析500μmol/L GDP-β－S,500min后被完全阻断;（2）外液中[Cl^-]从100mmol/L下降到5mmol/L时,在ATP（100μmol/L）作用下,ID2幅值随[Cl^-]的下降明显增大;（3）外液中无Na^ 时ID2幅值增高（P＜0．05）。结果表明,ID2是由P2Y受体所介导,通过G蛋白耦联及胞内信号转导而引起胸内Ca^2 依赖性Cl^-外流所致。