悦目金蛛卵袋的结构与组成

采用SEM和氨基酸自动分析仪对悦目金蛛(Argiope amoena)卵袋的结构、纤维组成和覆盖层的氨基酸组成进行了观察研究.结果表明悦目金蛛卵袋呈封闭的倒三角形,是由多种丝腺纺出的丝纤维形成的多个覆盖层构成的,从外向内分别为:金黄色大壶状腺框架丝,灰绿色、浅棕色、白色外覆盖层和棕色内覆盖层,其中外覆盖层是由6根大直径(约4.8-6.8 μm) 柱状腺丝纤维和一些小直径(约0.26-0.76 μm)纤维混合成的基本纤维束单元所折叠成的片层丝被构成的,小直径丝纤维的数量从灰绿色丝被(外)到白色丝被(内)逐渐增加,而内覆盖层仅仅是由6根大直径(约5.20 μm)柱状腺丝纤维构成的基本纤维束单元折叠而成.卵袋外覆盖层丝(白色绒状纤维除外)的甘氨酸、丙氨酸和丝氨酸等氨基酸的百分含量分别介于卵袋内覆盖层丝和葡萄状腺丝相对应氨基酸的百分含量之间,这一数据支持Foradori等关于卵袋小直径丝纤维是由葡萄状腺纺出的推论.悦目金蛛能调节卵袋层丝的颜色、直径、覆盖层丝纤维和氨基酸的组成,使卵袋获得各方面最优化的功能,为卵或若蛛提供支持和保护.     

The relationship between egg size and the duration of the facultative feeding period in marine invertebrate larvae

Feeding larvae of marine invertebrates fuel development from both endogenous egg energy and exogenous energy obtained from the planktonic environment. Although both sources of energy likely influence certain larval stages, only the effects of exogenous food have been well studied. Despite the lack of research on the effects of egg size on larval stages, investigators have hypothesized that egg size influences the duration of the facultative feeding stage—the stage in which larvae can feed but do not have to because development is still being fueled by egg energy. To test this hypothesis, we investigated six species of sand dollars with different sized eggs and quantified the duration of the larval facultative feeding period of each species by comparing when fed and starved larvae diverged in size. Regardless of whether phylogeny was taken into account, the duration of the facultative feeding period was positively correlated with egg size. We further determined that our conclusions were not sensitive to either our estimation of the duration of the facultative feeding period, or the branch lengths of the phylogeny we used. This relationship is likely a result of larger eggs being provisioned with more energy, and may affect how well larvae can cope with natural variability in food concentrations. Furthermore, our results support an assumption of a theoretical model developed to understand the evolution of different life-history strategies in marine invertebrate larvae, which suggests that this relationship has important evolutionary consequences.     

Egg weight variation in relation to egg mortality and starvation endurance of newly hatched larvae in some satyrid butterflies

Abstract. 1. Egg weights decrease over the oviposition period in all of the five satyrid butterflies that have been studied, viz Parage aegeria L., Lasiommata petropolitana F., L.maera L., L.megera L. and Lopingaachine Scop. However, within each species, no correlation was found between variation in egg weight and egg mortality in 30% relative humidity or the ability of newly hatched larvae to survive without food for 1–4 days. 2. Although egg weights of Papilio machaon L. were similar to the highest satyrid egg weights, newly hatched larvae of P.machaon showed significantly higher mortality after 1 day of starvation when compared to all of the satyrid larvae. This indicates that the capability of newly hatched larvae to endure starvation is not necessarily correlated with egg weight, and consequently that this character trait of satyrid larvae has been selected for (i.e. should be regarded as an adaptation). It is noteworthy that this ability of satyrid larvae to endure starvation is coupled with the habit of many satyrid butterflies not to deposit their eggs directly on the larval host plants. 3. In 30% relative humidity, egg mortality of L.achine was 100% and that of P.aegeria 29.2%, whereas that of the three Lasiommata spp. was significantly lower. At 100% relative humidity egg mortality in L.achine dropped to 8%. Since egg weights are higher in L.achine than in the other four satyrids this indicates that egg resistance to desiccation is not necessarily correlated with egg size (as might be expected due to the area to volume ratio which is negatively correlated with egg size). Thus the resistance to desiccation which is found in the three Lasiommata spp. should be regarded as an adaptation to the habit of these three butterflies to deposit their eggs in relatively dry microhabitats. This resistance to desiccation is not found in eggs of the two forest-dwelling satyrids P.aegeria and L.achine which lay their eggs in microhabitats where the relative humidity is high.     

The distribution of the transposable elementBari-1 in theDrosophila melanogaster andDrosophila simulans genomes

The distribution of the transposable elementBari-1 inD. melanogaster andD. simulans was examined by Southern blot analysis and byin situ hybridization in a large number of strains of different geographical origins and established at different times.Bari-1 copies mostly homogeneous in size and physical map are detected in all strains tested. Both inD. melanogaster and inD. simulans a relatively high level of intraspecific insertion site polymorphism is detectable, suggesting that in both speciesBari-1 is or has been actively transposing. The main difference between the two sibling species is the presence of a large tadem array of the element in a well-defined heterochromatic location of theD. melanogaster genome, whereas such a cluster is absent inD. simulans. The presence ofBari-1 elements with apparently identical physical maps in allD. melanogaster andD. simulans strains examined suggests thatBari-1 is not a recent introduction in the genome of themelanogaster complex. Structural analysis reveals unusual features that distinguish it from other inverted repeat transposons, whereas many aspects are similar to the widely distributedTc1 element ofC. elegans.     

Immunocytochemical evidence for the presence of two domains in the plasma membrane of sea urchin blastomeres

Summary The blastomeres of sea urchin embryos have two surface regions with different properties. Numerous microvilli are present in the apical surface region, while the baso-lateral surface region, either on adjoining adjacent cells or facing the blastocoel, is smooth. When blastomeres are isolated from embryos and stained with fluorescein-isothiocyanate-labelled anti-(egg surface) antibody (anti-ES) prepared against membranes isolated from fertilized eggs, the apical microvillous region fluoresces while the smooth region does not [Yazaki I (1984) Acta Embryol Morphol Exp 5∶3–22]. In order to study quantitatively the ‘bindability’ of the membrane in the two regions to anti-ES, immunoelectron microscopy was used. Blastomeres isolated from embryos ofHemicentrotus pulcherrimus at the eight-cell stage were treated with rabbit anti-ES serum or pre-immune serum and then with ferritin-conjugated goat anti-(rabbit IgG) for 10 min at 0°C, mainly before fixation. About 10 times (maximally 45 times) more ferritin particles were counted per contour length in the microvillous surface region than in the smooth surface region. These results suggest that the membrane of the blastomeres of sea urchin embryos is a mosaic of two different membrane territories: one represented by the microvillous surface originating from the unfertilized egg, which binds anti-ES, the other by the smooth surface newly organized after the first cleavage, which does not react with anti-ES. The mechanism of segregation of the membrane into these two regions is discussed.     

The ultrastructural organization of the isolated cortex in eggs ofNassarius reticulatus (Mollusca)

Summary We have studied the organization of the cortex in fertilized eggs ofNassarius reticulatus by examining rotary-shadowed whole mounts of isolated cortices in the transmission electron microscope. The following components were distinguished: (a) the plasma membrane, with clathrin-coated areas and coated pits, (b) microfilaments and microtubules, and (c) a tubulovesicular network of endoplasmic reticulum. Microfilaments were identified by labeling with heavy meromyosin, and microtubules with a monoclonal anti-tubulin antibody, using both immunofluorescence microscopy and immunogold labeling for transmission electron microscopy. The microfilaments are organized in a network parallel to and closely associated with the plasma membrane, with typical Y- and X-shaped intersections. The endoplasmic reticulum is associated with this microfilamentous lattice. The microtubules also run parallel to the plasma membrane, but they are located at a greater distance, as can be inferred from stereo images. In the uncleaved egg, numerous microtubules are present in the egg cortex. Shortly before polar lobe formation, at the onset of mitosis, the microtubules disappear almost entirely. They reappear again at the end of first cleavage, as the polar lobe is being resorbed. The synthesis of cortical microtubules at this stage appears to depend on the presence of microtubule-organizing centers in the animal hemisphere of the egg, since microtubules do not reappear in isolated polar lobes. Clathrin-coated areas are present in both the animal and vegetal hemisphere before polar lobe formation. During mitosis, the clathrin-coated plaques and pits are found almost exclusively in the animal hemisphere. After resorption of the polar lobe, at the two-cell stage, no clathrin-coated areas were found at all.     

Circadian rhythm of autophagy proteins in hippocampus is blunted by sleep fragmentation

Autophagy is essential for normal cellular survival and activity. Circadian rhythms of autophagy have been studied in several peripheral organs but not yet reported in the brain. Here, we measured the circadian rhythm of autophagy-related proteins in mouse hippocampus and tested the effect of sleep fragmentation (SF). Expressions of the autophagy-related proteins microtubule‐associated protein 1 light chain 3 (LC3) and beclin were determined by western blotting and immunohistochemistry. Both the hippocampal LC3 signal and the ratio of its lipid-conjugated form LC3-II to its cytosolic form LC3-I showed a 24 h rhythm. The peak was seen at ZT6 (1 pm) and the nadir at ZT16 (1 am). The LC3 immunoreactivity in hippocampal CA1 pyramidal neurons also distributed differently, with more diffuse cytoplasmic appearance at ZT16. Chronic SF had a mild effect to disrupt the 24 h rhythm of LC3 and beclin expression. Interestingly, a greater effect of SF was seen after 24 h of recovery sleep when LC3-II expression was attenuated at both the peak and trough of circadian activities. Overall, the results show for the first time that the hippocampus has a distinct rhythm of autophagy that can be altered by SF.     

The spatial arrangement of germ line cells in ovarian follicles of the mutantdicephalic inDrosophila melanogaster

Summary The pattern of intercellular connections between germ line cells has been studied in follicles of the mutantdicephalic (dic), which possess nurse cell clusters at both poles. Staining of follicles with a fluorescent rhodamine conjugate of phalloidin reveals ring canals and cell membranes and thus allows us to reconstruct the spatial organization of the follicle. Each germ line cell can be identified by the pattern of cell-cell connections which reflect the mitotic history of individual cells in the 16-cell cluster. The results indicate that in both wild-type anddicephalic cystocyte clusters one of the two cells with four ring canals normally becomes the pro-oocyte. However, in some follicles (dicephalic and wild-type) oocytes were found with fewer or more than four ring canals. Indic follicles, one or several nurse cells may become disconnected from the other cells during oocyte growth at stage 9–10. Such disconnected cells cannot later on empty their cytoplasm into the oocyte. This, in turn, might be of consequence for the determination of axial polarity of the embryo.     

LDFF, the large molecular weight DNA fragmentation factor, is responsible for the large molecular weight DNA degradation during apoptosis in Xenopus egg extracts

DNA degradation is a biochemical hallmark in apoptosis. It has been demonstrated in many cell types that there are two stages of DNA fragmentation during the apoptotic execution. In the early stage, chromatin DNA is cut into large molecular weight DNA fragments, although the responsible nuclease(s) has not been recognized. In the late stage, the chromatin DNA is cleaved further into short oligonucleosomal fragments by a well-characterized nuclease in apoptosis,the caspase-activated DNase (CAD/DFF40). In this study, we demonstrate that large molecular weight DNA fragmentation also occurs in Xenopus egg extracts in apoptosis. We show that the large molecular weight DNA fragmentation factor (LDFF) is not the Xenopus CAD homolog XCAD. LDFF is activated by caspase-3. The large molecular weight DNA fragmentation activity of LDFF is Mg^2 -dependent and Ca^2 -independent, can occur in both acidic and neutral pH conditions and can tolerate 45℃ treatment. These results indicate that LDFF in Xenopus egg extracts might be a new DNase (or DNases) responsible for the large DNA fragmentation.     

Muscle determinants in the ascidian egg are inactivated by UV irradiation and the inactivation is partially rescued by injection of maternal mRNAs

The ascidian egg contains cytoplasmic determinants that specify the fate of larval muscle cells. In a previous study, we developed an experimental system to identify the molecular nature of muscle determinants, in which unfertilized Ciona savignyi eggs were fragmented into four pieces by centrifugation. When inseminated, only nucleated fragments (red fragments) develop into partial embryos that only show differentiation of epidermal cells. One type of enucleated fragment (black fragment) has the remarkable ability to promote muscle differentiation when fused with red fragments. In the present study, using this experimental system, we investigated the molecular nature of muscle determinants. UV irradiation of black fragments suppressed the ability to promote expression of the muscle-specific protein, myosin heavy chain. The wavelength of UV light responsible for the inactivation (250–275 nm) suggested that UV-sensitive targets are nucleic acids. Injection of poly(A)⁺ RNA isolated from an un-irradiated black-fragment-rich fraction into UV-irradiated black fragments partially recovered the ability to promote the expression of myosin heavy chain protein. Poly(A)⁺ RNA from a red-fragment-rich fraction did not rescue the suppression of UV-irradiated black fragments. These results suggest that maternal mRNAs enriched in black fragments are closely associated with muscle determinants in the ascidian egg.     

The effects of egg position, egg mass size, substrate and biofouling on embryo mortality in the squid Sepioteuthis australis

Using a combination of laboratory and field investigations, this study examined embryo mortality in the southern calamary Sepioteuthis australis as a function of egg mass size, the substrate upon which the mass is attached, the position of the embryo within the mass, and the degree of biofouling. Egg mass size ranged from 2 to 1,241 egg strands, however most masses consisted of 200–299 strands. Small egg masses (<300 strands) were generally attached to soft-sediment vegetation (Amphibolis antarctica, Heterozostera tasmanica, Caulerpa sp.), whereas larger masses (>300 strands) were either securely attached to robust macroalgae holdfasts (Ecklonia sp., Marcocystis pyrifera, Sargassum sp.) or unattached. Rates of embryo mortality were highly variable ranging from 2 to 25%. Both laboratory and field results indicated a positive relationship between egg mass size and embryo mortality. Larger, unattached egg masses contained twice as many dead embryos than those securely attached to a substrate. Mortality rates were significantly affected by the embryos’ relative position within the mass and were highest in embryos located near the attachment point of the egg strand, within the interior of the mass, and in close contact with the substrate. This was attributed to the inability of the embryos to respire adequately and eliminate metabolic wastes. Biofouling did not strongly influence embryo mortality, but colonisation occurred in areas conducive to growth, photosynthesis, and respiration indicating ‘healthy’ regions within the mass.     

Experimental manipulation of egg quality in chickens: influence of albumen and yolk on the size and body composition of near-term embryos in a precocial bird

The importance of avian egg components in the determination of hatchling size and quality has yet to be fully evaluated. In the first experiment, 20% of the albumen and/or the yolk was removed from chicken eggs to determine the impact of each egg component on metabolism and various size measures in near-term embryos. Results show that metabolic rate, dry body mass, and internal organ mass are largely independent of egg composition. Removal of albumen resulted in a decrease in wet body mass corresponding to decreases in water content in the body and the yolk sac, and decreased tibiotarsus length. Removal of yolk resulted in no change in body mass, but decreases in both wet and dry yolk sac mass. In a second experiment, removal of 15% of either egg component led to reductions in hatchling mass similar to those observed in whole near-term embryos. Albumen, as the primary source of water in the egg, is the primary determinant of hatchling size and may influence hatchling success through size-related limiting factors. Differences in yolk content may influence neonatal quality as a nutritional supplement, but seem not to result in greater tissue formation during embryonic development. Accepted: 2 September 1997     

Induction of SCEs and DNA fragmentation in bovine peripheral lymphocytes by in vitro exposure to tolylfluanid-based fungicide

The potential for genotoxic and cytotoxic effects of tolylfluanid-based fungicide (50% active agent) was evaluated using sister chromatid exchange (SCE) and proliferation indices (PI) in cultured bovine peripheral lymphocytes. For the detection of possible genetic damage, DNA fragmentation assay was also applied. Bovine lymphocytes cultured for 72 h were treated with the fungicide at the final concentrations of 1.75, 3.5, 8.75, and 17.5 μg/mL for the last 24 and 48 h of culture without S9 metabolic activation, and during the last 2 h of culture with S9 metabolic activation. In the SCE assays no evidence for genotoxic activity of the fungicide was found in treatments of 24 h without and 2 h with S9. After the 24 h exposure to tolylfluanid, a weak decrease in the PI was observed. With the prolonged exposure time (48 h), dose dependence in the increase of SCE frequencies was observed. Moreover, after 48 h exposure slight fragmentation of DNA at the concentrations of 3.5 and 8.75 μg/mL was demonstrated. SCE quantification is the most widely used approach for the assessment of genotoxic/cytogenetic effects of chemical compounds. Positive results in the assay at 48 h exposure indicated a potential of the fungicide to increase frequency of chromosomal damage (replication injuries) that is the confirmation of early effect of exposure.     

Chemical nature of the egg shell in helminths. II. Mode of stabilization of egg shells of monogenetic trematodes

The chemical nature of the egg shells of the monogenetic trematodes Pseudomicrocotyle sp. and Pricea multae, parasitizing the gills of the marine fish Scomberomorus guttatus was studied with a view to understanding the mode of stabilization of the egg shells.The egg shells of the monogenetic trematodes appeared to contain dimers of tyrosine; a conclusion based on fluorescence properties, affinity to toluidine blue, methylene blue, and a positive test for aromatic amino acids. In addition, they are stabilized by -S-S- linkages as indicated by the positive reaction to tests for disulfides. This is also suggested by the shells being refractile to stain but which can be reversed after treatment with sodium thioglycollate.Absence of phenolic tanning in the egg shells of the monogenetic trematodes is indicated by the negative reaction to tests for quinones, the failure to develop color on incubation in catechol after heat treatment, and by the fluorescence in uv light.In these respects the egg shells of the monogenetic trematodes strongly resemble the egg shell of Fasciola hepatica.     

A comparison on the response to forest fragmentation by medium-sized Iberian carnivores in central Spain

We studied the use of forest fragments by five medium-sized carnivorespecies in 280 forest fragments on the two Iberian plateaus. We looked forindirect evidence (faeces, tracks, dens) of fragment use (occurrence) by thespecies and analysed whether occurrence could be related to four groups ofvariables: local (vegetation structure and patch size), landscape (distance topossible colonisation sources), regional (fragment location on the northern orsouthern plateau) and the vegetation type of the fragments. We analysed thedifferential response of species according to their life-history and behaviouraltraits. The relationship between use by each species and the factors studied wasanalysed using stepwise logistic regressions. Results indicate that threefactors are crucial to explain fragment use: fragment size, geographic locationand vegetation type. Large fragments are used more than smaller ones, thenorthern plateau is more suitable than the southern plateau, and holm oak andbroad-leaved oak forests are more used than pine woods. The effects andmagnitude varied slightly among species, depending on habitat requirements orlife-history traits. Data indicate that conservation strategies in fragmentedenvironments must take into account elements functioning at different spatialscales, and that it is essential to consider each case within a characteristicregional context.     

Structural change in the endoplasmic reticulum during the in situ development and in vitro fertilisation of wheat egg cells

The mechanism by which the sperm activates the egg cell of higher plants is largely unknown. Ca²⁺—implicated as a second messenger in the response of various plant cells to a wide range of stimuli—is a potential candidate for encoding the information brought by the sperm cell into the angiosperm egg. In higher plant cells the dominant calcium store appears to be the central vacuole; however, in the receptive wheat egg cell few vacuoles can be observed, thus it seems likely that the principal cell organelle performing a pivotal role in regulation of intracellular Ca²⁺ is the endoplasmic reticulum (ER). To examine this hypothesis, microinjection of the ER-specific fluorescent dicarbocyanine dye, 1,1-dihexadecyl-3,3,3,3-tetramethylindocarbocyanine perchlorate [DiIC₁₆(3)], and low-light level CCD technology were used. Following the injection of an oil drop saturated with DiI, structural changes occurring in the ER during the in planta maturation of the female gamete were visualised. The ER was identified by chlorotetracycline labelling to be the main calcium store in the wheat egg cell. Structural changes occurring in the ER during the in planta maturation of the wheat female gamete led to a polarised ER network in the receptive egg cell. Within 3 min of in vitro sperm-egg fusion, a rapid, transient loss of continuity of the ER occurred, which underlines the significance in fertilisation of the ER structure in the female gamete of wheat.     

Long-term intake of egg white hydrolysate attenuates the development of hypertension in spontaneously hypertensive rats

This paper evaluates the effect of the long-term intake of a hydrolysate of egg white with pepsin (HEW), with a potent angiotensin converting enzyme inhibitory activity, on the development of hypertension of spontaneously hypertensive rats (SHR). After being weaned, male 3-week-old SHR were randomly divided into five groups that were given until the 20th week of life the following drinking fluids: (1) tap water, (2) non-treated egg white 1 g/kg/day, (3) captopril 100 mg/kg/day, (4) HEW 0.5 g/kg/day, and (5) HEW 1 g/kg/day. From the 20th to 25th week of life, animals from all groups were given tap water. Systolic blood pressure (SBP) and diastolic blood pressure (DBP) were measured weekly in the rats, from the 6th to 25th week of life, by the tail cuff method. Development of hypertension was attenuated in the groups treated with captopril and HEW (P<0.001 vs. the group that drunk tap water). At the 20th week of life, the arterial blood pressure values of the different groups of rats were: tap water (SBP = 219.5 +/- 5.7, DBP = 167 +/- 3.7), non-treated egg white (SBP = 206.4 +/- 1.43, DBP = 166.4 +/- 4.9), captopril (SBP = 131.7 +/- 2.74, DBP = 91.5 +/- 1.62), HEW 0.5 g/kg/day (SBP = 182.9 +/- 4.64, DBP = 127.5 +/- 2.1) and HEW 1 g/kg/day (SBP = 177.7 +/- 4.72, DBP = 120.1 +/- 2.4). SBP and DBP increased in the treated SHR when the corresponding antihypertensive treatment was removed. In spite of this, SBP remained lower in the SHR that had received captopril and HEW than in the SHR of the control groups (P<0.05). The present results suggest that HEW could be used as a functional food with antihypertensive activity.