γ-微管蛋白在猪卵母细胞成熟和活化中的分布

微管蛋白(tubulin)是一蛋白质超家族,其中α-,β-微管蛋白是主要的微管蛋白,而γ-微管蛋白主要在微管组装中起作用. 我们利用蛋白质印迹和激光共聚焦技术研究了γ-微管蛋白在猪卵母细胞成熟、受精和活化中的分布. γ-微管蛋白存在于猪卵母细胞中,并且在减数分裂成熟各个时期的量保持不变. 它聚集在微管上,特别是中期纺锤体的两极和后末期的中板. 体外受精和孤雌活化后,γ-微管蛋白聚集在雌雄原核的周围.另外它也存在于精子的顶体帽和颈部.在早期卵裂中,γ-微管蛋白聚集在胚胎的细胞核周围.实验结果表明,γ-微管蛋白在猪卵母细胞、精子和胚胎的微管组装中起重要的调节作用,在猪受精过程中,精子和卵子都向受精卵贡献中心体物质.     

韩国黄蚜小蜂属分类研究(膜翅目:蚜小蜂科)

本文报道采自韩国的黄蚜小蜂属5种,包括1新种（Aphytis albus,sp．nov．)及4个韩国新记录种(Adiaspidis,A.japonicus,Aproclia和Avandenboschi),编制了雌成虫检索表。新种模式标本保存在韩国国立树木园和东北林业大学昆虫标本室。     

河南发现水蛛(蜘蛛目:水蛛科)

河南北部发现雌、雄水蛛Argyroneta aquatica.     

绵羊胞内单精子注射技术

本文的主要目标是建立绵羊胞内单精子注射技术(intracyioplasmic sperm injection,ICSI)。并且尝试了ICSI技术生产绵羊转基因胚胎的可能性。实验1比较了绵羊卵母细胞孤雌激活的3种化学方法。结果显示,Ionomvcin/3 h/6-dimethylaminopurine(6-DMAP)和Ionomycin激活胚的卵裂率(71.7%和91.8%)和囊胚率(17.4%和11%)显著(P<0.01)高于Ca~(2+)激活胚(18.4%和0)。Ionomycin/3 h/6-DMAP激活胚的囊胚发育形态和比例相对较高。实验2中,活精子注射至卵母细胞质后,用Ionomycin/3 h/6-DMAP激活,排出第二极体(PB2)的71枚ICSI胚胎用SOFaaBSA溶液培养,卵裂率为71.8%(51/71),显著(P<0.01)高于体外受精(41.4%,IVF)和阴性对照胚胎(30.2%,sham-ICSI)。培养7天后,sham-ICSI组没有囊胚生成;ICSI和IVF胚胎的囊胚率分别为7.0%(5/71)和16.1%(9/56),两者差异不显著(P>0.05)。这些ICSI囊胚在冷冻前后均能孵化,显示初步建立了绵羊ICSI技术。另外,我们探索了ICSI技术生产转基因胚胎的可能性,-20℃冻融1次的死精子与pEGFP-N1质粒共注射,33枚2-细胞期ICSI胚胎中,2枚可见GFP蛋白。其中1枚停止发育,另外1枚继续发育至16-细胞期,仍然可见GFP基因的表达。4枚解冻的ICSI囊胚手术移植给2只发情同期化受体绵羊,60天时,B超未见怀孕。本文初步结果表明,ICSI技术生产转基因绵羊有可能性,需深入研究。     

绵羊胞内单精子注射技术

In this study, the possibility of sheep transgenesis by intracytoplasmic sperm injection (ICSI) was assessed. In experiment 1, activation of ovine oocytes matured in vitro in preparation for ICSI has been investigated with 3.42 mmol/L Ca2+ treatment, ionomycin alone and ionomycin followed by 6-dimethylaminopurine (DMAP) after 3-h delay (group 1, 2 and 3, respectively). After activation, the oocytes were then cultured in SOFaaBSA medium. Cleavage rates were significantly (P<0.05) different among three groups (18.4%, 91.8% and 71.7%, respectively). In additional culture, no parthenotes in group 1, whereas 11% and 17.4% in group 2 and 3 developed to the blastocyst stage. Therefore we used the third activation method in the following ICSI tests. In experiment 2, development of ovine oocytes after ICSI was investigated. Thawed semen from two rams was separated by Percoll centrifugation and was used for ICSI or in vitro fertilization (IVF) trails. A total of 71.8% of oocytes reached the 2-cell stage following living sperm injection, which was significantly (p>0.05) different from those following IVF (41.4%) and sham-ICSI (30.2%). After seven days' culture, no sham-injected oocytes developed into the blastocyst stage, although 7% in ICSI and 16.1% in IVF-oocytes developed into the blastocyst stage, but there was no significant difference in ICSI and IVF groups (p>0.05). In the further study, the possibility of sheep transgenesis by ICSI was assessed. After coinjection of ovine oocytes matured in vitro with dead sperm cold to -20 degrees C and exogenous DNA encoding green fluorescent protein (GFP), seventy-three percent of coinjected oocytes developed to 2-cell stage (33/45) and two of them were transgene-expressing embryos. Among ten embryos at the 16-cell stage, all embryonic cells in one transgenic embryo still expressed GFP. Four coinjected blastocysts were thawed and transferred to the uterine of the two progesterone-synchronized recipient ewe. No pregnancies were detected on the 60th day. These results suggested sheep transgenic embryos could be produced by ICSI and further studies should be performed.     

银杏雌雄基因组DNA间的差异性分析

本研究应用RAPD技术,应用300个10bp随机单引物及200对随机双引物组合,检测了雌雄异株银杏基因组DNA的多态性。结果表明:雌雄基因组间具有极高的相似性,在检测到的3450个标记中,仅获得1个与银杏雄性基因组相关的RAPD标记。以该标记为探针,与雌雄银杏基因组DNA的Southern杂交分析,其杂交信号在两性之间表现为限制性片段长度多态性,该结果为寻找银杏早期性别鉴定的探针以及在细胞和分子水平进一步研究其性别问题奠定了基础。     

乳突寄螨雌螨描述(蜱螨亚纲: 革螨股: 寄螨科)

乳突寄螨 Parasitus mammillatus ( Berlese,1 90 4 )雌螨 ,在所查到的文献中 ,仅记载有生殖区、头盖和螯钳的形状 [1 ,2 ] ,无其它描述。国内根据丹东标本曾描述该螨雄螨和后若螨 [3] 。本文根据朝鲜标本详细描述其雌螨。文中测量单位为 μm,括号内为测量均值。图 1- 7　乳突寄螨 Parasitus mammillatus (Berlese)1.背面 ;2 .腹面 ;3.头盖 ;4 .螯钳 ;5 .颚角 ;6 .须肢 ;7.跗节 乳突寄螨 Parasitus mammillatus ( Berlese,1 90 4 )雌螨 (图 1 - 7)体黄色 ,卵圆形 ,长 71 2 - 793( 758) ,宽 4 0 2 - 483( 4 50 )。背板覆盖整个背面 ,前背…     

葛藤雌雄蕊发育的研究

通过光学显微镜和扫描电镜对葛藤的雌雄蕊发育进行观察,结果表明：葛藤雌雄蕊的发育及结构基本上符合蝶形花科的胚胎学特征,其花粉粒为2－细胞型,胚囊发育为蓼型。     

北美香柏雌球果的发育

用扫描电镜(SEM)观察了北美香柏 Thuja occidentalis 雌球果的发育过程。在北京,北美香柏的雌球果是在八月初由营养芽转变而来,雌球果一般有4~6对苞片,中间2~3对可育,每一苞片腋部着生两枚胚珠,在可育苞片腋部最先观察到一扁平的隆起,并在其上分化出两个胚珠原基,接着分化出珠被和珠心,最后形成扁平而两侧对称的胚珠。在北美香柏雌球果发育过程中,约一半的雌球果在2~3对可育苞片中位于下面的1~2对的腋部产生3个胚珠原基,中间一个较小,并在以后的发育中逐渐退化。由此推测北美香柏的雌球果可能是由祖先类群中每一苞片具多于2个胚珠的雌球果演化而来。在光镜下对雌球果维管系统的观察发现,传粉前幼小雌球果的苞片内仅有一束维管束,传粉后随着苞片基部的居间生长,有4—8束维管束在苞片内形成,但是新发育的维管束木质部和韧皮部相对位置与正常叶性器官一致,这与在以往报道的柏科植物成熟雌球果的苞片中均有反向维管束的发育不同。北美香柏雌球果早期发育和维管束分析结果支持傅德志和杨亲二提出的解释裸子植物生殖器官形态演化的“苞鳞-种鳞复合体”理论。关键词北美香柏;雌球果的发育;胚珠分化;SEM