长江华溪蟹卵子发生的细胞化学研究

利用细胞化学的方法，对长江华溪蟹卵子发生过程中，核酸、蛋白质、糖类及脂类的变化进行了研究，结果表明：核酸从卵原细胞到卵母细胞的发育进程中逐渐分散；蛋白质的量不断增加；脂类物质也在慢慢积累，伴随着发育，在成熟的卵母细胞质内充满ＰＡＳ阳性反应的卵黄颗粒。     

日本血吸虫的组织化学研究 Ⅱ.脂类、脂酶和非特异性酯酶的组织化学证明

1.对从感染日本血吸虫的小白鼠体内取出的发育未成熟的幼虫、性刚成熟的成虫、充分成熟的成虫以及从豚鼠体内取出的成虫或经离体培养后的虫休内脂类物质的分布世行了研究,并对成虫体内的脂酶和非特异性酯酶作了初步观察。 2.发育1周和2周的日本血吸虫幼虫其体内未能查见脂类物质,发育3周性刚成熟的成虫含有少量脂类颗粒,而在发育5周后充分成熟的虫体内则沉着大量的脂类物质,主要分布于雄虫实质组织和雌虫的卵黄腺及虫卵的卵黄细胞内。 3.成虫在葡萄糖-台氏液中离体培养3天后其体内含有大量脂类,并在若干原先未能查见脂类的肠管上皮细胞和肠腔内呈现脂类阳性颗粒。但在无糖台氏液中“饥饿”20小时后,未见虫体内形态上可见的脂类阳性反应有何显著减弱。 4.根据所用显示脂类反应的特异性,可以辨认出在雄虫实质组织内是中性脂肪而在雌虫的卵黄细胞中主要为磷脂和少量中性脂肪。 5.应用Tween法或α-萘酚醋酸酯法未能发现日本血吸虫成虫体内有脂酶或非特异性酯酶的存在。 6.对日本血吸虫成虫体内所沉着的脂类物质的来源和生理意义进行了讨论。     

短额负蝗卵子发生过程中糖复合物的动态分布

以生物素标记的凝集素(UEA-I、SBA、PNA)为探针，利用凝集素组织化学方法对短额负蝗(Atractomorpha sinensis)卵子发生过程中滤泡细胞和卵母细胞内糖复合物的分布进行了定位研究。结果表明，在卵子发生的各期滤泡细胞和卵母细胞中没有UEA-I受体的表达，SBA和PNA受体以不同的分布模式呈阶段性表达。两者首次出现于卵母细胞生长期， 随后PNA受体消失，SBA受体大量表达；在卵黄形成期前期SBA受体和重新出现的PNA受体表达于卵黄颗粒形成部位，卵黄形成期后期两者均为阴性表达；成熟卵子中两种受体又以不同程度重新出现于卵黄膜。两种受体在滤泡细胞内均大量表达 提示，N-乙酰半乳糖胺和半乳糖-β-(1，3)半乳糖胺复合物的修饰和变化与卵母细胞的发育、卵黄物质的形成及滤泡细胞的增殖分化密切相关，卵黄膜上的糖复合物可能与精卵识别有关。     

泥螺卵子发生的超微结构研究

利用透射电镜观察了泥螺卵子发生过程。结果表明 ,泥螺的卵子发生可划分为卵原细胞、卵黄发生早期、卵黄发生中期及卵黄发生后期卵母细胞 4个时期。卵原细胞核大而圆 ,胞质内分布有少量的线粒体和高尔基囊泡 ,细胞表面具微绒毛。卵黄发生早期的卵母细胞 ,胞质中各类细胞器发达 ,并出现数量较多的类朦胧子。卵黄发生中期的卵母细胞胞体迅速增大 ,核伸出伪足状突起 ,卵质中各种细胞器活动活跃 ,并参与形成卵黄粒和脂滴。此期还可观察到卵母细胞与滤泡细胞间的物质交换现象。卵黄发生后期的卵母细胞体积增至最大 ,细胞器数量减少。本文就卵黄发生前后卵母细胞内部构造的变化、意义及滤泡细胞与卵母细胞蛋白来源间的关系作了探讨     

泥螺卵子发生的超微结构研究

利用透射电镜观察了泥螺卵子发生过程。结果表明,泥螺的卵子发生可划分为卵原细胞、卵黄发生早期、卵黄发生中期及卵黄发生后期卵母细胞4个时期。卵原细胞核大而圆,胞质内分布有少量的线粒体和高尔基囊泡,细胞表面具微绒毛。卵黄发生早期的卵母细胞,胞质中各类细胞器发达,并出现数量较多的类朦子。卵黄发生中期的卵母细胞胞体迅速增大,核伸出伪足状突出,卵质中各种细胞器活动活跃,并参与形成卵黄粒和脂滴。此期还可观察到卵母细胞与滤泡细胞间的物质交换现象。卵黄发生后期的卵母细胞体积增至最大,细胞器数量减少。本文就卵黄发生前后卵母细胞内部构造的变化、意义及滤泡细胞与卵母细胞蛋白来源间的关系作了探讨。     

短额负蝗卵子发生过程中糖复合物的动态分布

以生物素标记的凝集素(UEA-I、SBA、PNA)为探针,利用凝集素组织化学方法对短额负蝗(Atracto-morphasinensis)卵子发生过程中滤泡细胞和卵母细胞内糖复合物的分布进行了定位研究。结果表明,在卵子发生的各期滤泡细胞和卵母细胞中没有UEA-I受体的表达,SBA和PNA受体以不同的分布模式呈阶段性表达。两者首次出现于卵母细胞生长期,随后PNA受体消失,SBA受体大量表达;在卵黄形成期前期SBA受体和重新出现的PNA受体表达于卵黄颗粒形成部位,卵黄形成期后期两者均为阴性表达;成熟卵子中两种受体又以不同程度重新出现于卵黄膜。两种受体在滤泡细胞内均大量表达。提示,N-乙酰半乳糖胺和半乳糖-β-(1,3)半乳糖胺复合物的修饰和变化与卵母细胞的发育、卵黄物质的形成及滤泡细胞的增殖分化密切相关,卵黄膜上的糖复合物可能与精卵识别有关。     

犬首鮈卵子发生过程及成熟卵膜形态结构观察

摘要：通过组织切片和扫描电镜技术对犬首鮈卵子发生过程及成熟卵膜表面结构进行观察。结果显示,犬首鮈卵子发生过程可以分为Ⅰ-Ⅵ时相:第Ⅰ时相卵母细胞主要有卵原细胞构成;第Ⅱ时相卵母细胞外层出现滤泡细胞;皮质液泡和卵黄颗粒分别发生在第Ⅲ、Ⅳ时相;第Ⅴ时相卵母细胞为成熟的卵子,富含丰富的卵黄和少量皮质液泡;第Ⅵ时相卵母细胞退化进而被卵巢吸收。卵膜孔呈漏斗状,卵孔区域类型属于降旋混合系统。犬首鮈成熟卵膜表层具有特殊的柱状体结构,高度为18.3~30.6um,平均分布密度为0.15个/um2,分析可能为该物种或生态类型的卵膜结构特征,推测对产沉粘性卵鱼类受精卵孵化、胚胎发育起重要作用。     

棉铃虫卵巢形态与卵子发生过程观察

害虫发生高峰期、 发生量的准确预测和田间防治适期的确定与种群雌虫卵巢结构及卵子发生过程密切相关。为了明确棉铃虫Helicoverpa armigera卵巢结构及卵子发生过程, 本研究利用光学体视显微镜和透射电子显微镜, 对棉铃虫成虫卵巢管和卵子的超微结构进行了研究, 并确定了发育级别划分标准。结果表明: 根据卵巢的形状、 卵的产生过程、 卵黄沉积情况等将棉铃虫卵巢发育程度分为6个级别, 即发育初期(0级)、 卵黄沉积前期(Ⅰ级)、 卵黄沉积期(Ⅱ级)、 成熟待产期(Ⅲ级)、 产卵盛期(Ⅳ级)和产卵末期(Ⅴ级)。根据卵子发生过程中滋养细胞、 卵母细胞的变化, 将卵子发生期分为3个阶段： 卵黄发生前期、 卵黄发生期和卵黄成熟期。本研究首次对棉铃虫的卵子发生进行电子显微观察, 并完善了棉铃虫卵巢发育的分级标准, 为进一步研究棉铃虫的生殖发育机理提供了理论参考, 对田间棉铃虫种群发生期和发生量的预测预报也有重要的实践参考价值。     

不同体重瘤背石磺性腺发育规律

利用组织学方法研究了瘤背石磺体重(2—28g)与性腺发育、性腺指数与肝胰腺指数或卵黄腺指数间的关系,不同体重瘤背石磺性腺内各期生殖细胞的组成与比例以及瘤背石磺卵子和精子发生的规律。结果表明,(1)瘤背石磺的性腺指数有随体重增加而增加的特点:10g以上个体性腺指数达到最高且基本无变化;不同体重瘤背石磺性腺指数与肝胰腺指数和卵黄腺指数有明显的正相关性(P<0.05);(2)6g以下组的瘤背石磺性腺滤泡管内未发现有雌性生殖细胞,6g以上组的性腺滤泡管内雄性与雌性生殖细胞并存;(3)所有瘤背石磺个体性腺内均有精子分布,6g以下个体雄性生殖细胞组成以次级精母细胞为主,而6g以上个体则以精子为主;6g以上组的雌性生殖细胞成熟程度随体重增加有明显增加,其中6—8g以卵原细胞为主(57%),8—10g开始出现外源性卵黄合成期的卵母细胞,10—14g时的外源性卵黄合成期的卵母细胞约为69%,且开始出现成熟卵母细胞。(4)卵子发生共经历6期:分别为卵原细胞期、卵黄合成前卵母细胞期、内源性卵黄合成期、外源性卵黄合成期、近成熟期和成熟卵母细胞期,成熟卵母细胞直径约为(59.36±3.88)μm。精子发生经历精原细胞、初级精母细胞、次级精母细胞、精子细胞和精子共5个阶段,精子长约(52.44±20.65)μm。石磺体重与性腺发育程度密切相关,10g以上的个体可做为亲本使用。     

昆虫卵黄原蛋白和卵黄发生

近年来,昆虫卵黄发生(vitellogenesis)的机理和控制的研究是昆虫生理学中最活跃的研究领域之一。昆虫具有强大的繁殖力,能在短时间内产生大量含有卵黄的成熟卵。卵黄是由多种大分子组成的复合物,包括蛋白质、脂类和碳水化合物,其中蛋白质是主要成分。卵黄发生就是各种卵黄物质的形成及其在卵母细胞中的沉积。这是卵细胞发育成熟的必要前提。     

OVARIAN NUTRITIONAL RESOURCES DURING THE REPRODUCTIVE CYCLE OF THE HEMATOPHAGOUS DIPETALOGASTER MAXIMA (HEMIPTERA: REDUVIIDAE): FOCUS ON LIPID METABOLISM

In this study, we have analyzed the changes of the ovarian nutritional resources in Dipetalogaster maxima at representative days of the reproductive cycle: previtellogenesis, vitellogenesis, as well as fasting‐induced early and late atresia. As expected, the amounts of ovarian lipids, proteins, and glycogen increased significantly from previtellogenesis to vitellogenesis and then, diminished during atresia. However, lipids and protein stores found at the atretic stages were higher in comparison to those registered at previtellogenesis. Specific lipid staining of ovarian tissue sections evidenced remarkable changes in the shape, size, and distribution of lipid droplets throughout the reproductive cycle. The role of lipophorin (Lp) as a yolk protein precursor was analyzed by co‐injecting Lp‐OG (where OG is Oregon Green) and Lp‐DiI (where DiI is 1,10‐dioctadecyl‐3,3,30,30‐tetramethylindocarbocyanine) to follow the entire particle, demonstrating that both probes colocalized mainly in the yolk bodies of vitellogenic oocytes. Immunofluorescence assays also showed that Lp was associated to yolk bodies, supporting its endocytic pathway during vitellogenesis. The involvement of Lp in lipid delivery to oocytes was investigated in vivo by co‐injecting fluorescent probes to follow the fate of the entire particle (Lp‐DiI) and its lipid cargo (Lp‐Bodipy‐FA). Lp‐DiI was readily incorporated by vitellogenic oocytes and no lipoprotein uptake was observed in terminal follicles of ovaries at atretic stages. Bodipy‐FA was promptly transferred to vitellogenic oocytes and, to a much lesser extent, to previtellogenic follicles and to oocytes of ovarian tissue at atretic stages. Colocalization of Lp‐DiI and Lp‐Bodipy‐FA inside yolk bodies indicated the relevance of Lp in the buildup of lipid and protein oocyte stores during vitellogenesis.     

Changes in lipid droplet localization during embryogenesis of the silkworm, Bombyx mori

Lipid droplets are considered one of the most important energy sources in lepidopteran eggs during late embryogenesis, but the process of their incorporation into the embryo is as yet unknown. The present study focused on the process of transition of lipid droplets from the extraembryonic yolk to the embryo of the silkworm Bombyx mori, using morphological and biochemical approaches. The morphological study revealed that the incorporation of lipid droplets from the extraembryonic yolk into the embryo occurs at three points and in three different ways during the development of the embryo. Some lipid droplets were translocated directly from the extraembryonic yolk to the embryo before the blastokinesis stage. However, the majority of lipid droplets together with the other components of the extraembryonic yolk were incorporated in the embryo via both morphogenetic inclusion during dorsal closure and ingestion of the extraembyonic yolk by the developing caterpillar prior to hatching. Similar results were obtained from the biochemical study. Thus, we propose that there are three steps in the incorporation of lipid droplets from the extraembryonic yolk into the embryo. In addition, morphological and biochemical data concerning the total amount of lipid droplets in the egg suggested that lipid droplets were mainly consumed during late embryogenesis, seeming to synchronize with tracheal development.     

银鲫受精卵中卵黄粒和线粒体的超微结构变化

电镜观察到雌性银鲫肝脏中正在形成的卵黄物质具有晶形主体结构,银鲫卵的卵黄粒内无晶形主体,第一次卵裂前的受精卵卵黄粒内有两种形式的空泡。一种是一些中空的小空泡;另一种是一个大的空泡,泡内含有线粒体和颗粒状的核糖体,泡的边缘有片层状的类脂物。受精卵中的线粒体内部结构多样,有些含有颗粒状物,有些含有片层状的类脂物。     

A CYTOLOGICAL STUDY OF THE CENTRIFUGED WHOLE, HALF, AND QUARTER EGGS OF THE SEA URCHIN ARBACIA PUNCTULATA

While the ooplasmic components of centrifuged eggs of Arbacia punctulata do not stratify in homogeneous layers, we have obtained the following strata beginning with the centripetal end: lipid droplets, pronucleus, clear zone, mitochondria, yolk, and pigment. Whereas mitochondria may be found mingled with yolk bodies, we have never observed lipid droplets nor pigment bodies among any of the other inclusions. The so-called clear zone contains a heterogeneous population of inclusions: annulate lamellae, heavy bodies, Golgi complexes, and rod-containing vacuoles. The peripheral cortical granules of immature (germinal vesicle stage) and of mature eggs are not dislodged from the cortical ooplasm with the centrifugal force utilized. When the eggs are treated with urethane, prior to centrifugation, the cortical granules of mature eggs abandon their peripheral position. Further centrifugation of the initially stratified eggs produces nucleated and nonnucleated halves and the centrifugation of the halves results in quarters. The cytology of the halves and quarters is discussed. The halves and quarters have been activated with either sperm or hypertonic sea water. With the exception of the nucleated halves, we were unable to obtain plutei larvae from the other fractions (red halves and quarters). We believe that the lack of development of the various fragments is a function of the balance of particular inclusions necessary for differentiation.     

Vitellogenesis in the crayfish Rhynchocinetes typus: role of hepatopancreas in lipid yolk biosynthesis

Biochemical studies in lipid composition of yolk granules from the crayfish Rhynchocinetes typus is shown to be composed mainly of cholesterol and phospholipids. Thin layer chromatography analysis demonstrates phosphatidyl-choline, -ethanolamine and -serine as its major phospholipidic components. In vivo labeling experiments using 32P-orthophosphoric acid suggest that two of the major yolk phospholipid components, phosphatidyl -choline and -serine could be synthesized in the hepatopancreas and subsequently transported via hemolymph to the growing oocyte.     

Effects of Added CeCl<Subscript>3</Subscript> on Resistance of Fifth-Instar Larvae of Silkworm to <Emphasis Type="Italic">Bombyx mori</Emphasis> Nucleopolyhedrovirus Infection

The aim of this study was to investigate the influence of mineral sources on broiler breeders and their offsprings. Broiler breeding hens were fed with diets containing either organic or inorganic trace minerals at equal levels, i.e., (1) control group was fed with basal diet supplemented with inorganic trace minerals; (2) OZ group was fed with organic Zn instead of sulfate; and (3) OTM group was fed with organic Cu, Mn, Zn, and Se instead of inorganic sources. Results indicated that OTM supplementation decreased plasma cholesterol and triglyceride and increased yolk triglyceride via increasing high-density lipid protein cholesterol and decreasing low-density lipid protein cholesterol and very low-density lipid protein (VLDL) in plasma. OZ diets decreased plasma cholesterol and triglyceride mainly by reducing VLDL concentration. For control group, increased lipid concentrations resulted in increased lipid peroxidation in serum and malondialdehyde retention in yolk. Zn retention was not affected. Otherwise, OZ diet was observed to decrease Cu in yolk and albumen. While for OTM group, albumen Cu, albumen Se, and hepatic Se of hatched chicks were increased, but yolk Cu was decreased. Moreover, organic mineral supplementations improved broilers’ growth performance. In conclusion, organic mineral supplementation in breeders’ diets protected breeders from lipid peroxidation, increased egg nutrition retention, and benefit for growth of broilers.     

EGG COMPOSITION AND HATCHLING PRECOCITY IN SEABIRDS*

This paper presents analyses of the composition (shell, yolk, albumen) and of the quality (lipid, protein and water contents) of yolk and albumen of eggs of 25 species of seabirds belonging to the Sphenisciformes, Procellariiformes, Pelecaniformes, and Lari of the Charadriiformes. These data are compared with those published for species in the same orders and suborders, and the combined seabird data are compared with published data for non-seabirds. Variation in the proportionate weight of eggshell is not related to hatchling precocity but is probably due to adaptive differences in nest substratum and the agility of the incubating birds. Lipid and protein contents of yolk, and water content of albumen, are uniform in eggs of all examined species. Protein levels of albumen and water content of yolk and of entire egg contents vary greatly between taxa. Contrary to previous claims there is no simple relationship between egg composition and hatchling precocity. Differences in egg composition will probably only be clearly understood when the nutritional needs of the embryo, and of the hatchling in the time before it regularly obtains food, are known.     

THE ROUTE OF ENTRY AND LOCALIZATION OF BLOOD PROTEINS IN THE OOCYTES OF SATURNIID MOTHS

The oocytes of saturniid moths take up proteins selectively from the blood. The distribution of blood proteins in the ovary during protein uptake was investigated by staining 2 µ sections of freeze-dried ovaries with fluorescein-labeled antibodies. The results indicate that blood proteins occur primarily in the intercellular spaces of the follicle cell layer, in association with a brush border at the surface of the oocyte, and within the oocyte in the yolk spheres. That proteins derived from the blood are associated with the yolk spheres was confirmed by isolating these bodies and showing that lysis, which can be induced by any of a number of mechanical means, causes them to release immunologically defined proteins known to be derived from the blood. That the level of blood proteins in the cytoplasm is low relatively to that in the yolk spheres was confirmed by the observation that the yellow pigments associated with several blood proteins, although conspicuous in the yolk spheres, are not visible in the translucent layer of centrifuged oocytes. From these and previous physiological observations, it is proposed that blood proteins reach the surface of the oocyte by an intercellular route, that they combine with some component of the brush border, and that they are transformed into yolk spheres by a process akin to pinocytosis.     

蓖麻蚕卵组织蛋白酶B和组织蛋白酶D活化机制及作用(英)

本文研究了蓖麻蚕卵内组织蛋白酶B和组织蛋白酶D在酸性条件下的活化机理和在卵黄蛋白水解中的作用。活性检测结果表明,酸处理用以使蛋白酶活化,但SDS-聚丙烯酰胺凝胶电泳实验表明酸化没有使蛋白酶分子量降低。推测酸化可能通过改变蛋白酶构象从而使蛋白酶活化。两种蛋白酶之间未表现出相互修饰导致分子量变化,也未表现出明显的相互促进活性的作用。蛋白酶水解产物分子量的不同,说明两种蛋白酶作用位点不同。组织蛋白酶B的水解产物分子量较小而组织蛋白酶D的水解产物分子量大,说明两种蛋白酶在体内可能作用于蛋白质水解的不同阶段或作用于不同的蛋白质。免疫扩散结果显示,抗蓖麻蚕卵蛋白酶血清可以识别柞蚕和棉铃虫卵内的成分,表明其它一些鳞翅目昆虫卵内可能存在相似的蛋白酶。