NBS修饰~(241)Trp对33kD外周蛋白与PSⅡ的结合及放氧活性恢复的影响

高等植物光系统Ⅱ（ＰＳⅡ）中膜结合的３３ｋＤ外周蛋白对锰分子簇的稳定及ＰＳⅡ高放氧活力的维持具有重要意义。本文通过ＮＢＳ对菠菜３３ｋＤ蛋白中唯一的色氨酸残基（近Ｃ端２４１Ｔｒｐ）的修饰研究了该残基在光合放氧系统中的作用。结果表明２４１Ｔｒｐ位于３３ｋＤ蛋白内部的疏水区。修饰后的３３ｋＤ外周蛋白对ＰＳⅡ颗粒的重组能力减弱。部分重组上经ＮＢＳ修饰的３３ｋＤ蛋白的ＰＳⅡ颗粒放氧活力得不到恢复。表明２４１Ｔｒｐ对３３ｋＤ蛋白与ＰＳⅡ的结合及功能的实现有着特殊意义。     

猪卵母细胞的体外受精及多精受精

对用于猪体外受精(IVF)的研究方法和技术,如传统的液滴IVF、透明带下注射精子受精(SUZI)、卵母细胞质内单精注射受精(ICSI)及细管IVF等进行了简述。与其它动物相比,进行猪卵的体外受精研究,多精受精现象特别明显。大量的研究表明,猪卵的多精受精不但与其品种特性有关,而且与卵母细胞成熟的程度、透明带的异常、受精时获能精子的浓度、输卵管分泌物、受精液蛋白添加成分、NaHCO3浓度、咖啡因、pH值以及温度等因素密切相关。     

NBS修饰^241Trp对33kD外周蛋白与PSⅡ的结合及放氧活 …

高等植物光系统Ⅱ（ＰＳⅡ）中膜结合的３３ｋＤ外周蛋白对锰分子的稳定及ＰＳⅡ高放氧活力的维持具有重要意义。本文通过ＮＢＳ对菠菜３３ｋＤ蛋白中唯一的色氨酸残基（近Ｃ端＾２４１Ｔｒｐ）的修饰研究了该残基在光合放氧系统中的作用。     

一种与受精有关的人精子膜蛋白（BS—17）的分离纯化

本文盐分级分离,Ｍｏｎｏ Ｑ ＦＰＬＣ及ＳＤＳ－聚烯酰胺胶电泳（ＳＤＳ－ＰＡＧＥ）等方法从人精子ＣＨＡＰＳ抽提液中分离纯化出一种与不育病血清中抗精子抗体发生特异反应的ＢＳ－１７人精子膜蛋白。该蛋白为一糖蛋白,分子量为１７．５５±２．１５ｋＤ,等电点为５．６５,中性己糖含是为１６．６７％。在人精子上主要分布于顶体区域,不同于已有报导的人精子膜蛋白。在体外实验中抗ＢＳ－１７多克隆血清可以显著影响人精子     

蓖麻蚕卵天冬氨酸蛋白酶纯化及性质

采用硫酸铵沉淀,离子交换层析和离子交换凝胶过滤法,从蓖麻蚕卵母细胞中纯化出一种蛋白酸疼。该蛋白酶活性能被天冬氨酸蛋白酶特性异性抑制剂胃肽抑制,初步鉴定该酶为天冬氨酸蛋白酶,经ＳＤＳ－聚丙烯配角安凝胶电泳测得电泳测得分子量为９０ｋＤ,由凝胶过滤估计分子量在３６０ｋＤ,推测该酶由四亚基组成。     

豆薯种子中两种蛋白质的分离纯化及其性质研究

豆薯（Ｐａｃｈｙｒｒｈｉｚｕｓｅｒｏｓｕｓ）种子经磷酸盐缓冲液抽提,Ｓ－ＳｅｐｈａｒｏｓｅＦａｓｔＦｌｏｗ柱,ＤＥ－５２纤维素柱和ＳｅｐｈａｄｅｘＧ－７５柱层析,提取出两种高纯度的蛋白成分,命名为ＰａｃｈｙｒｉｎＩ和ＩＩ,ＳＤＳ－ＰＡＧＥ测得其分子量分别为３３ｋＤ和１４．５ｋＤ,但ＨＰＬＣ分子筛的结果显示ＰａｃｈｙｒｉｎⅡ的分子量为２８ｋＤ,无论在还原条件下,还是在非还原条件下,ＰａｃｈｙｒｉｎＩ     

增强UV—B辐射对菜豆蛋白质代谢的影响

模拟研究了温室条件下兰州地区臭氧层减薄２５ ％ 时增强的ＵＶＢ（２８０ ～３２０ ｎｍ） 辐射对菜豆（ Ｐｈａｓｅｏｌｕｓｖｕｌｇａｒｉｓ Ｌ．） 蛋白质代谢的影响。菜豆幼苗期,ＵＶＢ辐射（ＵＶＢＢＥ：１６ ｋＪ·ｍ －２·ｄ－１） 促进菜豆叶片中蛋白质的合成;菜豆生长后期,ＵＶＢ辐射则抑制蛋白质的合成,降低叶片中可溶性蛋白质的含量,促进蛋白水解酶活性的上升和总游离氨基酸的积累。ＳＤＳＰＡＧＥ分析表明,ＵＶＢ辐射使幼苗期菜豆叶片中８８ ｋＤ、７６ ｋＤ、４２ ｋＤ、３５ ｋＤ、３３ ｋＤ 多肽的含量上升;而在菜豆生长后期,ＵＶＢ辐射使大分子量多肽（７６ ｋＤ 以上） 的含量迅速下降,１０～１４ ｋＤ、２９ ｋＤ、３３ ｋＤ、３５ｋＤ的多肽含量上升。蛋白质代谢的这种变化可能是植物的一种适应性反应     

蛋白激酶在卵母细胞减数分裂和受精中的作用

脊椎动物卵母细胞的减数分裂和受精过程受到多种蛋白激酶的调节。近年来对于卵母细胞成熟、活化和受精的分子机制研究取得了长足进步 ,发现促成熟因子 (MPF)和促分裂原活化蛋白激酶 (MAPK)是调节卵母细胞细胞周期的关键分子 ,二者的激活和失活导致了减数分裂的恢复、阻滞和完成。许多蛋白激酶通过调节MPF和MAPK活性来影响减数分裂。Polo like激酶活化MPF ,Mos激活MAPK而启动成熟分裂并维持中期阻滞。CaMKII通过泛素途径灭活MPF使卵突破MII期阻滞。另外 ,p90 rsk作为MAPK的下游分子参与减数分裂调节 ,蛋白激酶C(PKC)诱导皮质颗粒排放并抑制MAPK激活 ,酪氨酸蛋白激酶家族成员介导受精诱发的Ca2 释放。这些蛋白激酶的协同作用推动了卵母细胞正常的成熟与受精     

蛋白激酶C在小鼠卵母细胞体外成熟和受精中的作用

蛋白激酶是一类重要的丝／苏氨酸蛋白激酶。本实验以小鼠为实验动物,研究了PKC在卵母细胞体外成熟、活化和受精中的可能作用,及两种PKC亚型在卵母细胞中的定位。PKC激活剂PMA可以阻止GV期卵母细胞在体外恢复减数分裂,该作用可被PKC抑制剂CalphostinC抵消,但不能被PLCγ抑制剂U73122或PKCδ专一性抑制剂Rottlerin所克服。Western印迹显示PKCα和βI在卵母细胞发育过程中恒量表达。激光共聚焦显微术研究发现,受精或受到活化刺激后PKCα转位到卵母细胞膜上,同时皮质颗粒排放,说明PKCα可能参与调节卵皮质反应。本实验首次在小鼠中研究了PLCγ与受精的关系,发现不存在PKC对PLCγ的正反馈调节。此外,本研究还对小鼠卵巢中对PKCα和βI进行了蛋白定位研究。     

蛋白激酶C在小鼠卵母细胞体外成熟和受精中的作用

蛋白激酶是一类重要的丝/苏氨酸蛋白激酶。本实验以小鼠为实验动物,研究了PKC在卵母细胞体外成熟、活化和受精中的可能作用,及两种PKC亚型在卵母细胞中的定位。PKC激活剂PMA可以阻止CV期卵母细胞在体外恢复减数分裂,该作用可被PKC抑制剂calphostin C抵消,但不能被PLCγ抑制剂U73122或PKCδ专一性抑制剂Rottlerin所克服。Western印迹显示PKCα和βⅠ在卵母细胞发育过程中恒量表达。激光共聚焦显微术研究发现,受精或受到活化刺激后PKCα转位到卵母细胞膜上,同时皮质颗粒排放,说明PKCα可能参与调节卵皮质反应。本实验首次在小鼠中研究了PLCγ与受精的关系,发现不存在PKC对PLCγ的正反馈调节。此外,本研究还对小鼠卵巢中对PKCα和βⅠ进行了蛋白定位研究。     

绿蟾蜍繁殖期间输卵管变化的组织学研究

本文研究了绿蟾蜍繁殖期间输卵管组织结构变化的规律。产卵前，输卵管壁最厚，固有层中的单管状腺充满胶质；产卵时，粘膜层形成细长的皱襞，固有层中的腺体分泌胶质，形成包裹卵子的卵胶膜；产卵后，输卵管壁变薄，腺体细胞缩小，但腺体间的结缔组织隔膜增厚，结缔组织可能对腺体细胞的修复起重要作用。     

Changes in femoral robusticity and porosity during the reproductive cycle of the female alligator (Alligator mississippiensis)

To help understand the nature of skeletal changes during the reproductive cycle of the female alligator, we compared femoral robusticity (density) and porosity of cross-sections from the midshafts of femora from the following groups of female alligators: (1) immature; (2) pre-ovulatory; (3) postovulatory with soft-shelled oviducal eggs; (4) post-ovulatory with hard-shelled oviducal eggs; (5) post-ovulatory with eggs in the nest; (6) post-ovulatory with hatched eggs; and (7) mature, quiescent. Femora from alligators with eggs in the nest were significantly less robust (dense) than those of the other groups except those with hard-shelled oviducal eggs. Cross sections from the midshaft of femora from alligators with eggs in the nest were significantly more porous than those from all the other groups. The results indicated that calcium was mobilized from the femoral shaft shortly before eggs were laid and that femoral density returned to normal levels for mature alligators 1–2 months after egg-laying.     

Localization of plasminogen in the extracellular matrix of hamster eggs: exogenous activation by streptokinase

The plasminogen activator (PA)/plasminogen/plasmin proteolytic system has begun to be taken into account in the fertilization process. In this study, we demonstrated the presence of plasminogen in the extracellular matrix (ECM) of hamster oocytes by indirect immunofluorescence and immunoperoxidase assays using human anti-plasminogen. Plasminogen appeared first on the zona pellucida (ZP) of ovarian oocytes and later on the plasma membrane (PM) of oviducal eggs. This would suggest that oviducal oocytes modulate the expression of plasminogen binding sites on the PM. Human plasminogen as well as that of other species, known to be activated by streptokinase (SK), is rapidly converted to a plasmin-SK complex. We demonstrated the rapid formation of a SK-plasminogen complex that yields plasmin in the blood plasma of hamsters. Both the in vivo and in vitro SK treatment of eggs from superovulated female hamsters caused a decreased in the ZP dissolution time (ZPdt), probably either due to the proteolytic effect of plasmin or due to the SK-Plasminogen. Extracellular proteolysis assays carried out on agar-casein plates confirmed the proteolytic activity of SK-incubated eggs; the controls, on the contrary, failed to display a halo. These studies show that (1) superovulated hamster eggs contain plasminogen in their ECM, (2) oviducal eggs exhibit plasminogen on their PMs, indicating the presence of their corresponding binding sites, (3) in hamsters, SK, a non-enzymatic exogenous protein would be capable of activating ECM plasminogen to plasmin, and (4) the complex SK-plasminogen and/or the plasmin are capable of changing the ZPdt with alpha-chymotrypsin.     

RE-EXAMINATION OF FERTILIZATION INHIBITION STUDIES WITH ANTI-FROG-EGG-JELLY ANTIBODIES: EXPERIMENTS WITH NON-PRECIPITATING ANTIBODIES

Antisera were prepared in rabbits against oviducal and egg-jellies of the frog, Rana japonica . Gamma-globulin fractions from the antisera were degraded to a univalent, non-precipitating form by a papain digestion-reduction procedure. Agar diffusion analyses proved that the digested antibody fragments were inhibitory to the precipitin reaction of undigested, multivalent antibodies with jelly antigens. The treatment of unfertilized eggs with undigested, multivalent antibodies resulted in a significant loss of egg-fertilizability. In contrast, treatment with the univalent fragments of antibodies did not affect egg-fertilizability, similarly to the treatment with both univalent and multivalent γ-globulins from control, non-immune sera. Fertilization was inhibited in large measure when unfertilized eggs were subjected to a dual treatment with univalent antibodies and γ-globulins from anti-rabbit γ-globulin sheep serum. The inhibition of fertilization in the above experiments was always accompanied by the formation of a precipitation layer at the surface of the jelly envelopes. It is concluded that the failure of fertilization in the multivalent antibody-treated eggs results from a secondary effect rather than a specific blocking of a sperm-jelly interaction essential for fertilization.     

On gonadic maturation and reproductive strategy in deep-sea benthic octopus Graneledone macrotyla

The new information reported in this paper is based on five maturing and mature females of the large-tuberculate octopus Graneledone macrotyla. These specimens were caught in bottom trawl surveys ATLANTIS 2009 (February 24 to April 1, 2009) and ATLANTIS 2010 (March 9 to April 5, 2010) carried out off the Argentinean Economic Exclusive Zone. Capture depth ranged from 475 to 921 m and sea bottom temperature between 2.8 and 3.1 °C. Development of the complex ovary, oviducts, and oviducal glands during gonadic maturation is described. The absence of spermathecae in the oviducal glands and the presence of fertilized eggs inside the ovary suggested that fertilization took place within the ovary. Histological techniques showed the presence of four types of oocytes. Maturing oocyte size–frequency distribution was polymodal. Fluorescence reaction showed that atresia occurred in both early and later oocyte maturation stages. Atresia affected 48–55 % of the initial number of oocytes. The maximum observed potential fecundity was estimated at 250–300 eggs. G. macrotyla showed a group-synchronous ovulation pattern, regulative atresia, and a batching spawning pattern with a few egg batches spawned intermittently over an extended period of spawning.     

Properties of the Cortical Granule Lectin Isolated from Xenopus Eggs

The cortical granule lectin that participates in forming the fertilization layer in Xenopus laevis was isolated and partially characterized. About 400 μg of lectin was purified from 5 mg of crude exudate by chromatography on Sepharose 6B and Concanavalin A-conjugated Sepharose 4B columns and electrophoretic separation on polyacrylamide gel. The lectin has a molecular weight of 550 Kd and is composed of two species of polypeptides (46 Kd and 42 Kd). The lectin gave a single precipitin line against material in the prefertilization layer in an agglutination reaction on an agarose plate. The agglutination reaction involved D-galactoside residues and metal ions. The lectin formed an electron-dense layer on the outer surface of the vitelline coat of oviducal eggs covered with the prefertilization layer, but on the outer surface of jelly layer, not on that of the vitelline coat of jellied eggs. Although the jelly could be agglutinated by the lectin, the possibility that the jelly layer is the site of fertilization layer formation was excluded by the fact that the prefertilization layer is the first to meet the cortical granule lectin during normal fertilization.     

MAP kinase links the fertilization signal transduction pathway to the G1/S-phase transition in starfish eggs.

The mechanism by which fertilization initiates S-phase in the zygote is examined by manipulating the activity of MAP kinase in mature starfish eggs. These unfertilized eggs, which are arrested at G1-phase after the completion of meiosis, have high MAP kinase activity but undetectable cdc2 kinase activity. Either fertilization or inhibition of protein synthesis causes a decrease in MAP kinase activity, which is followed by DNA synthesis. Inactivation of MAP kinase with its specific phosphatase, CL100, initiates DNA synthesis in the absence of fertilization, while constitutive activation of MAP kinase with MEK represses the initiation of DNA synthesis following fertilization. Thus, in unfertilized mature starfish eggs, a capacity for DNA replication is already acquired, but entry into S-phase is negatively regulated by MAP kinase activity that is supported by a continuously synthesized protein(s) but not by cdc2 kinase. Upon fertilization, downregulation of MAP kinase activity is necessary and sufficient for triggering the G1/S-phase transition.     

Progesterone secretion by the post-ovulatory rat cumulus oophorus

This study was designed to determine whether cumulus oophorus cells secrete progesterone. Immature PMS-primed, hCG-treated rats were used. Their cumuli were isolated from pre-ovulatory (no hCG) and peri-ovulatory (after hCG) follicles, and from post-ovulatory oviducal ampullae. Treatment with hCG increased progesterone secretion by almost two-and-one-half-fold. It was speculated that the cumulus oophorus secretion of progesterone modifies the accumulation of fluid in the ampulla, thus providing the medium in which fertilization takes place.     

革胡子鲶受精过程的扫描电镜观察

应用扫描电镜观察和描述了革胡子鲶成熟卵和精子的形态、卵壳膜的表面结构和形态、受精孔的位置和结构、精子入卵过程的程序和变化。讨论了精子入卵过程及精孔细胞在解体之后可能转变为一种能够吸引精子在精孔区聚集的“受精素”物质等问题。