受精鸡蛋卵黄中DNA和RNA的分布

通过对受精未孵育鸡蛋卵黄的不同部位加以考查,发现DNA和RNA在卵黄中的分布是广泛存在的;采用荧光试剂Hoechst33258和RiboGreen对DNA和RNA进行定量分析,结果表明：DNA在不同部位卵黄中的分布比较均匀,且主要存在于卵黄颗粒中,而RNA的含量在胚下表层卵黄、侧面和植物极中有较大差异.     

鸡卵黄DNA的存在、提取和特性分析

利用免疫细胞化学技术证实了鸡蛋卵黄球中DNA的存在 .利用Ficoll 40 0密度梯度离心纯化卵黄球并提取了DNA .限制性内切酶分析表明所得卵黄DNA是序列不均一的DNA分子群 .MspⅠ和HpaⅡ分析显示该DNA的甲基化程度极低 .Southern杂交表明卵黄DNA含有基因组DNA的一小部分序列 .并认为卵黄DNA是一种独特的有可能在鸡胚早期发育中发挥功能的DNA .     

染色质在非洲爪蟾卵的无细胞系统中形成组装核的研究

从大鼠肝、鸡肝提取了染色质并从受精未孵育鸡蛋胚下表层卵黄颗粒提取了染色质,在荧光显微镜和电子显微镜下观察了这些染色质在无细胞系统中形成的组装核;研究二阶钙镁离子对形成组装核的影响。在有ＡＴＰ再生系统存在的无细胞系统中,围绕染色质能形成与间期细胞核相似的组装核;二价钙离子不利于形成组装核,而二价镁离子是形成组装核所必须的,但过高浓度的镁离子对形成组装核有抑制作用。     

鸡胚的多原条

鸡是多精受精的动物,通常孵一个鸡蛋发育出一只雏鸡。在鸡胚发育过程中有时会出现二个或三个原条,曾经有人报导过,这样的鸡卵如能顺利地继续发育下去,可能会孵出2只或3只雏鸡来。但是发生3个以上原条的情况则不多见了。我们在制作鸡胚标本时,曾获得一个有9个原条的标本,它们均匀分布于卵黄的表面,打开卵壳之后,先将整个卵黄     

云南油杉原胚发育过程中组织化学的变化

应用组织化学方法对云南油杉(Keteleeria evelyniana Mast)受精前后,原胚及幼胚形成早期细胞内的DNA、RNA、碱性蛋白质,酸性蛋白质及多糖物质进行了观察。结果表明,DNA在卵核受精前后为孚尔根弱正反应。在原胚及早期幼胚发育过程中,胚原细胞核DNA含量恢复正常,RNA及酸性蛋白质含量均较丰富,特别是在胚原细胞内。新细胞质中碱性蛋白质呈负反应,而DNA、RNA、酸性蛋白质及多糖物质均呈现正反应。     

鸡胚卵黄球能否演变为胚胎细胞?

Ο.Б.勒柏辛斯卡娅认为跌入于胚下腔中的卵黄球能够演变为胚胎的内胚层细胞和进入于内外二胚层之间的卵黄球能够演变为血岛的这一假说近年来在细胞学和胚胎学界引起了激烈的争论. 我们从1954年春天开始研究这个问题,工作可分三方面:(1)分别观察鸡胚正常发生过程中的内胚层和血岛的形成过程和卵黄球的变化——包括鸡胚和鸭胚从未孵到孵育20天各期的胚胎和卵黄囊切片,采用各种主要的细胞学上的染色方法和卵黄球的活体染色观察.(2)卵黄球的离体和活体培养——培养材料包括未孵及孵育各期的胚盘下卵黄球.离体培养     

小鼠体内外受精植入前胚胎全基因组甲基化模式比较

为考察体外受精、操作及培养环境对体外受精的小鼠植入前胚胎全基因组DNA甲基化模式的影响,本研究以体内受精的植入前胚胎作为对照,采用间接免疫荧光法检测小鼠体内外受精植入前胚胎基因组DNA甲基化模式.实验结果表明,体外受精各期植入前胚胎呈现出与之相应时期的体内受精植入前胚胎不同的DNA甲基化模式和水平,原核期甲基化水平较高,2-4-、8-细胞期明显降低,而桑葚胚和囊胚期又略有升高.各期体外受精植入前胚胎的基因组DNA甲基化水平都比同时期体内受精胚胎的甲基化水平低.本实验结果部分显示了体外受精、操作及培养环境可能对正常的DNA甲基化模式产生影响,造成体外受精植入前胚胎甲基化模式异常.     

家蚕卵裂核第一次分裂的方位观察

卵裂在胚胎学上有一定的规律,Wilson(1892)曾从几何学的角度,说明卵裂面对于卵轴或胚轴的位置关系,裂球对于正体位置关系的卵裂类型,主要取决于成胚原形质和卵黄的分配。家蚕卵的类型属于中黄卵,在卵黄膜内几乎全是卵黄集中分配的地方,只有在动物极的漏斗状范围内,卵黄分配稀少。卵的成熟分裂、受精和卵裂都开始在卵的动物极内(图1),然后引向植物极。     

新疆特克斯河斑重唇鱼胚胎发育和胚后发育观察北大核心CSCD

为保护伊犁河水系特克斯河特有鱼类斑重唇鱼(Distychus maculates)种质资源,探究斑重唇鱼胚胎发育与胚后发育规律。2021年5月,对斑重唇鱼开展人工繁殖,观察其胚胎发育和胚后发育过程,记录各发育阶段形态特征。水温15.0~16.0℃条件下,斑重唇鱼胚胎发育历经受精、卵裂、囊胚、原肠胚、神经胚、器官形成和孵化出膜7个阶段,经过2969.830℃·h破膜而出。斑重唇鱼成熟卵子淡黄色,卵径(2.75±0.24)mm(n=20),卵受精35 min后卵周隙最大,为(3.46±0.16)mm(n=20),其中卵黄约占体积的3/5。胚后发育过程中,仔鱼全长(L_(T),mm)和出膜天数(D)关系式:L_(T)=0.50D+9.15(R^(2)=0.90);卵黄囊全长(L_(Y))和出膜天数(D)关系式:L_(Y)=﹣0.11D^(2)+7.61(R^(2)=0.76);仔鱼全长(LT)和卵黄囊体积(L_(P))关系式:LT=﹣2.35LP+19.96(R2=0.88);卵黄囊全长(LY)和卵黄囊体积(L_(P))关系式:L_(Y)=0.49LP+5.34(R^(2)=0.68);卵黄囊体积(L_(P))和出膜天数(D)关系式:L_(P)=0.002D^(2)﹣0.24D+4.67(R^(2)=0.98)。本研究通过特克斯河斑重唇鱼人工繁殖实验,观察其胚胎和胚后发育特征,丰富斑重唇鱼早期生活史资料,为特有鱼类苗种培育提供理论数据,进一步为种质资源保护和开发奠定基础。     

多胚水稻品系APⅣ不同类型胶囊的受精及其胚胎形成

通过GMA半薄切片技术对APⅣ不同类型水稻(Oryza sativa L.)胚囊的受精及其胚胎发育的研究表明,APⅣ中5-2-l型胚囊的3个卵细胞在少数情况下都可受精并发育形成3个胚;但多数情况只有 1个或2个卵细胞受精发育成1个胚或2个胚。6-2-0型和5-3-0型胚囊多个卵受精频率都很低。由此证明APIV多胚是来自如5-2-1型胚囊的多卵卵器胚囊多个卵细胞都受精的结果,其中3胚来自3个卵细胞受精发育,2胚来自2个卵细胞受精发育。双套结构胚囊受精最为复杂,多数情况是受精不正常,只有少数子房大、小胚囊中的卵细胞都能正常受精。大胚囊中的卵细胞受精发育可能是形成所谓中位胚(远离珠孔端胚)的主要原因。     

Presence, isolation and characterization of yolk DNA from chicken eggs

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Hypotonic buffer induces meiosis and formation of anucleate cytoplasmic islands in the egg of the two-spotted cricket Gryllus bimaculatus

In insects, egg activation is known to occur in vivo and independently of fertilization, but its mechanisms are poorly understood. To gain understanding of these mechanisms, an attempt was made to activate the egg of Gryllus bimaculatus in vitro. It was found that meiosis resumed and was completed in unfertilized eggs treated with hypotonic buffer. Early developmental processes in activated, unfertilized eggs were investigated and compared with those in fertilized eggs. Mitosis did not progress, resulting in formation of anucleate cytoplasmic islands (pseudoenergids). Development in the activated, unfertilized eggs stopped at this stage and both yolk subdivision and cellularization did not occur. To elucidate the role of the nucleus in the developmental process to the syncytial stage in fertilized eggs, eggs were treated with aphidicolin to inhibit DNA polymerization. It was found that pseudoenergids also formed in these aphidicolin-treated fertilized eggs. These results demonstrate that pseudoenergids can increase in number independently of nuclei, suggesting that the cytoplasm rather than the nucleus plays the primary role in development to the syncytial stage in G. bimaculatus.     

Changes in elemental composition of fertilized and unfertilized northern pike (Esox lucius) eggs incubated in buffer with and without dimethyl sulfoxide: An X-ray microanalysis study

Fertilized and unfertilized eggs from the northern pike (Esox lucius) were incubated 2 hr in buffer with 0 and 10% (v/v) dimethyl sulfoxide and then quickly frozen in the wells of aluminum blocks submerged in liquid nitrogen. Control eggs and ovarian fluid were similarly frozen immediately after collection. The frozen eggs were sectioned, freeze dried, mounted on stubs, and carbon coated. X-ray microanalysis was used to determine changes in element levels and dimethyl sulfoxide (Me₂SO) penetration in the zona radiata, cytoplasm, cortical alveoli, and egg yolk. Unfertilized eggs incubated without Me₂SO showed decreased levels of Na, Cl, and K in the zona radiata; fertilized eggs, incubated without Me₂SO showed decreased levels of Na, P, and Cl in the zona radiata and increased levels of K in the cytoplasm; unfertilized eggs, incubated with 10% Me₂SO showed decreased Na and Cl in the zona radiata, decreased K in the cytoplasm and increased K in the cortical alveoli; fertilized eggs incubated in buffer with 10% Me₂SO showed decreased levels of Na, P, Cl, and K (zona radiata), P, Cl, and K (cytoplasm), Na (yolk), and increased Cl in the yolk (all P<.01). Me₂SO (v/v) levels reached 1.5-3.1% in the zona radiata, 0-3.2% in cytoplasm, 2.3-8.7% in cortical alveoli, and 0-1.6% in the yolk. Unfertilized eggs showed more Me₂SO penetration than fertilized eggs.     

Immunoelectron Microscopic Demonstration of Cortical Granule Lectins in Coelomic, Unfertilized and Fertilized Eggs of Xenopus laevis

Immunoelectron microscopic studies demonstrated cortical granule lectins (CGLs) in coelomic, unfertilized and fertilized eggs of Xenopus laevis . An antiserum raised against purified cortical granule lectin 1 specifically reacted with the CGLs in immunoblotting and agar diffusion tests. When ultrathin sections were treated with the antiserum and protein A-gold solution, gold particles, indicating antigenic sites, were seen over cortical granules of coelomic and unfertilized eggs, and over the perivitelline space, the vitelline coat and the condensed region of the fertilization layer of fertilized eggs. The pre-fertilization layer immediately adjacent to the outer margin of the vitelline coat in unfertilized eggs was free from gold particles. These observations suggest that released CGLs permeate through the vitelline coat of fertilized eggs and interact with the pre-fertilization layer mainly at the outer margin of the vitelline coat, resulting in formation of the fertilization layer which acts as a block to polyspermy.     

Adaptation of cultured human lymphoblasts to growth in citrulline

DNA synthesis is initiated in unfertilized sea urchin eggs (Strongylocentrotus purpuratus and Lytechinus pictus) by exposing them to NH₄OH-sea water (ordinary sea water titrated to pH 9–9.1 with NH₄OH). The eggs are considered to be unfertilized eggs by visual and electro-biological criteria and because they can later be fertilized and then do give visible and electrobiological fertilization reactions. The incorporation of ³H-thymidine proceeds in rounds, the magnitude increasing in successive rounds. It is also reported that the treatment with NH₄OH activates the uptake of thymidine by the eggs, although the internal thymidine builds up more slowly in unfertilized eggs treated with NH₄OH than it does in fertilized eggs. The magnitude of the incorporation of exogenously supplied labelled thymidine into DNA is lower in the NH₄OH-treated unfertilized eggs than in normal fertilized eggs. This difference is not attributed to differences in the amount of DNA synthesized and the explanation is sought in thymidine uptake and nucleotide pathways.     

Actin in triton-treated cortical preparations of unfertilized and fertilized sea urchin eggs

Triton-treated cortical fragments of unfertilized and fertilized sea urchin eggs prepared in the presence of greater than or equal to 5 mM EGTA contain 15-30% of the total egg actin. However, actin filaments are not readily apparent by electron microscopy on the cortical fragments of unfertilized eggs but are numerous on those of fertilized eggs. The majority of the actin associated with cortical fragments of unfertilized eggs is solubilized by dialysis against a low ionic strength buffer at pH 7.5. This soluble actin preparation (less than 50% pure actin) does not form proper filaments in 0.1 M KCl and 3 mM MgCl2, whereas actin purified from this preparation does, as judged by electron microscopy. Optical diffraction analysis reveals that these purified actin filaments have helical parameters very similar to those of muscle actin. Furthermore, the properties of the purified actin with regard to activation of myosin ATPase are similar to those of actin from other cell types. The possibility that actin is maintained in a nonfilamentous form on the inner surface of the unfertilized egg plasma membrane and is induced to assemble upon fertilization is discussed.     

Preparation of plasma membranes from fertilized sea urchin eggs

A new method is presented for preparation of highly purified plasma membranes from fertilized sea urchin eggs. The purified plasma membranes are in vesicle form and are highly enriched in ouabain inhibitable, Na+/K+ ATPase activity. Analysis of membrane proteins by sodium dodecyl sulfate-gel electrophoresis indicates that several high-molecular-weight proteins characteristic of plasma membranes from unfertilized eggs are absent in plasma membranes from fertilized eggs.     

Interspecific fusion of sea urchin eggs: Surface events and cytoplasmic mixing

Here we describe a new method of fusing fertilized and unfertilized sea urchin eggs. After removal of materials peripheral to the plasma membranes, the plycation polyarginine is used to induce the eggs to adhere to one another. The subsequent incubation of the eggs in an artificial sea water with a slightly lower osmolarity and a higher calcium content than normal sea water results in successful fusion. We demonstrate with scanning electron microscopy (SEM) that our artificial sea water serves to smooth the plasma membranes—a characteristic which we find to be a requirement in order for fusion to occur. We also demonstrate that our fusion procedure can be used to identify and follow the development of hybrids composed of eggs of different histories. Using species whose cytoplasms are distinguishable by light microscopy, Strongylocentrotus purpuratus and Lytechlnus pictus, we consider the mixing of cytoplasms using yolk granules as markers. We find that if two unfertilized eggs are fused, the species-specific yolk particles are free to mix. In contrast, we observe no mixing of the yolk particles in either fertilized-unfertilized or fertilized-fertilized interspecies hybrids until the time of mitosis. At that time, the yolk particles are free to move around the region containing the mitotic figure(s). However, mixing of the cytoplasmic yolk granules is never completed during this brief period. Even after cytokinesis takes place, one can see the segregation of the yolk granules in the cytoplasms of the two species.     

A direct effect of some rifamycin derivatives on the morphology of mammalian mitochondria

Protein synthesis has been investigated in cell-free preparations from mature ovarian oocytes, unfertilized and fertilized eggs, and early embryos of Drosophila melanogaster. Preparations from unfertilized eggs have a specific activity that is 5- to 6-fold higher than the activity of fractions from ovarian oocytes. There is an additional small increase in activity of preparations from fertilized eggs. The specific activity that is rapidly attained in the fertilized egg remains essentially constant for 2 to 2.5 h after fertilization, decreases sharply during blastoderm formation, and again increases during gastrulation. The activities of unfertilized eggs decline slightly during the first 2 h after oviposition, and then decrease more sharply. About 35 % of the ribosomes in preparations from both unfertilized and fertilized eggs sediment in the polyribosome region of sucrose density gradients, whereas no polyribosomes could be detected in preparations from ovarian oocytes. In both ovarian oocytes and fertilized eggs, less than 1 % of the ribosome populations were present as subunits. Additional ribonucleoprotein material of buoyant densities different from those of ribosomal subunits or ribosomes was found throughout the sucrose gradients. About 3.5 % of the ribosomes were found to be membrane-bound in preparations from both unfertilized and fertilized eggs.     

Taxol inhibits the nuclear movements during fertilization and induces asters in unfertilized sea urchin eggs

Taxol blocks the migrations of the sperm and egg nuclei in fertilized eggs and induces asters in unfertilized eggs of the sea urchins Lytechinus variegatus and Arbacia punctulata. Video recordings of eggs inseminated in 10 microM taxol demonstrate that sperm incorporation and sperm tail motility are unaffected, that the sperm aster formed is unusually pronounced, and that the migration of the egg nucleus and pronuclear centration are inhibited. The huge monopolar aster persists for at least 6 h; cleavage attempts and nuclear cycles are observed. Colcemid (10 microM) disassembles both the large taxol-stabilized sperm aster in fertilized eggs and the numerous asters induced in unfertilized eggs. Antitubulin immunofluorescence microscopy demonstrates that in fertilized eggs all microtubules are within the prominent sperm aster. Within 15 min of treatment with 10 microM taxol, unfertilized eggs develop numerous (greater than 25) asters de novo. Transmission electron microscopy of unfertilized eggs reveals the presence of microtubule bundles that do not emanate from centrioles but rather from osmiophilic foci or, at times, the nuclear envelope. Taxol-treated eggs are not activated as judged by the lack of DNA synthesis, nuclear or chromosome cycles, and the cortical reaction. These results indicate that: (a) taxol prevents the normal cycles of microtubule assembly and disassembly observed during development; (b) microtubule disassembly is required for the nuclear movements during fertilization; (c) taxol induces microtubules in unfertilized eggs; and (d) nucleation centers other than centrioles and kinetochores exist within unfertilized eggs; these presumptive microtubule organizing centers appear idle in the presence of the sperm centrioles.