Fertilization of the eggs ofCynops pyrrhogaster (japanese newt) after immersion in water

Summary Cynops pyrrhogaster oviducal eggs with and without jelly envelopes (jelly egg and dejellied egg respectively) were immersed in water, and then inseminated artificially. After 1 h of immersion in water, more than half the dejellied eggs were fertilized and developed, but no jelly eggs developed. The rapid decrease in the ability of jelly eggs to be fertilized after immersion in water is not due to a deficiency in the egg. Our results make it clear that hydrated jelly envelopes prevent the eggs from fertilizing. The ability of the egg to be fertilized decreases for a long time in water and this decrease proceeds more slowly in De Boer's solution or Holtfreter's balanced salt solution than in water.     

The role of jelly coats in sperm-egg encounters, fertilization success, and selection on egg size in broadcast spawners

Sperm limitation may be an important selective force influencing gamete traits such as egg size. The relatively inexpensive extracellular structures surrounding many marine invertebrate eggs might serve to enhance collision rates without the added cost of increasing the egg cell. However, despite decades of research, the effects of extracellular structures on fertilization have not been conclusively documented. Here, using the sea urchin Lytechinus variegatus, we remove jelly coats from eggs, and we quantify sperm collisions to eggs with jelly coats, eggs without jelly coats, and inert plastic beads. We also quantify fertilization success in both egg treatment groups. We find that sperm-egg collision rates increase as a function of sperm concentration and target size and that sperm are not chemotactically attracted to eggs nor to jelly coats in this species. In fertilization assays, the presence of the jelly coat is correlated with a significant but smaller-than-expected improvement in fertilization success. A pair of optimality models predict that, despite the large difference in the energetic value of egg contents and jelly material, the presence of the jelly coat does not diminish selection for larger egg cell size when sperm are limiting.     

THE EFFECT OF SOLUBLE EGG JELLY ON THE FERTILIZABILITY OF ACID-DEJELLIED SEA URCHIN EGGS*

Acid-dejellied Lytechinus pictus eggs bind few sperm and show decreased fertilizability. Addition of solubilized egg jelly increases sperm binding and fertilizability, presumably by increasing the frequency of the acrosome reaction. However, dejellied Strongylocentrotus purpuratus bind more sperm and show increased fertilizability in the complete absence of soluble egg jelly. Addition of soluble egg jelly greatly decreases fertilizability in S. purpuratus. Such species differences may be the basis for the controversy between Lillie and Tyler on the one hand, who believed that egg jelly increased egg fertilizability; while Loeb and Hagström on the other hand, believed jelly had no effect on, or actually decreased egg fertilizability. ¹²⁵I-labeling of dejellied S. purpuratus egg surfaces and immunofluorescent studies show that egg jelly persists on the surfaces of acid-dejellied eggs. Egg jelly appears to be a non-removable component of the vitelline layer of this species.     

Maternal energy investment in eggs and jelly coats surrounding eggs of the echinoid Arbacia punctulata

In free-spawning marine invertebrates, the amount of maternal energy that is invested in each egg has profound implications for all life-history stages of the offspring. The eggs of echinoids are freely spawned into the water and are surrounded by several structurally complex extracellular layers. These extracellular layers, or jelly coats, do not contribute energy to embryonic development but must impose an energy cost on the production of each egg. The investment of maternal energy reserves in the jelly coats of echinoid eggs may have important implications for the number of eggs that can be produced (i.e., fecundity) and the amount of energy that can be invested in each egg. We estimated the degree to which maternal energy is invested in the jelly coats surrounding eggs of the echinoid Arbacia punctulata. Estimates were derived from measurements of the amount of energy contained in the combined eggs and jelly coats, and in the eggs alone. The amount of energy contained in A. punctulata eggs ranged from 2.70 to 5.53 x 10(-4) J egg(-1). The amount of energy contained in the jelly coats ranged from 0.13 to 0.48 x 10(-4) J jelly coat(-1). The mean concentration of energy in the eggs was 2.15 mm(-3) and 0.29 J mm(-3) in the jelly coats. These results indicate that between 3% and 11% (mean = 7%) of the total energy invested in each A. punctulata egg is partitioned to the jelly coat alone. A significant positive relationship was found between the volumes of the jelly coats and the amount of energy they contained. Based on this relationship and an analysis of differences in the size of jelly coats between echinoid species, we suggest that the degree to which energy is invested in jelly coats may vary among echinoid species and is therefore likely to be an important life-history characteristic of these organisms.     

The jelly envelopes and fertilization of eggs of the newt, Notophthalmus viridescens

Fertilization in Notophthalmus viridescens is internal and involves passage of the sperm through five layers of egg jelly (J5-J1, from outermost to innermost), each of which is secreted by a discrete region of the oviduct. Polyspermy is normal. Passage of the sperm through the jelly and into the egg was studied by a technique of artificial insemination similar to natural insemination, in that undiluted fluid from the vas deferens was applied directly to eggs with various layers of jelly present, followed by flooding with water three to five minutes later. In general, successful fertilization increased as the number of jelly layers increased; jellyless coelomic eggs were not fertilizable. Sperm passage through the jelly and into the egg usually occurs within one to three minutes. Upon hydration of the jelly, barriers to sperm penetration develop in layers J5 and J3. Changes in the egg jelly thus seem to be involved in the restriction of polyspermy to a low level.     

The relations of the plasma membrane,vitelline membrane,and jelly in the egg of Nereis limbata

1. The problem of the relation of the plasma membrane to the extraneous coats and cortex of the Nereis egg is discussed in the light of the observations of Lillie, Chambers, and Novikoff. 2. Evidence obtained from experiments with the centrifuge, and by treating eggs with alkaline sodium chloride, indicates that the plasma membrane of the unfertilized egg is external to the jelly precursor granules of the cortex. 3. Experiments with alkaline sodium chloride indicate that the perivitelline space of the fertilized egg is extraovular after jelly extrusion is complete. 4. The cortical behavior (membrane elevation) of the Nereis egg in alkaline sodium chloride and the cortical response (jelly extrusion) following activation of the egg in normal fertilization or parthenogenesis are attributed largely to the properties of the jelly, and presumably, to its reactions with calcium and hydroxyl ions.     

Structural features of the abalone egg extracellular matrix and its role in gamete interaction during fertilization

Abalone eggs are surrounded by a complex extracellular coat that contains three distinct elements: the jelly layer, the vitelline envelope, and the egg surface coat. In this study we used light and electron microscopy to describe these three elements in the red abalone (Haliotis rufescens) and ascribe function to each based on their interactions with sperm. The jelly coat is a spongy matrix that lies at the outermost margin of the egg and consists of variably sized fibers. Sperm pass through this layer with their acrosomes intact and then go on to bind to the vitelline envelope. The vitelline envelope is a multilamellar fibrous layer that appears to trigger the acrosome reaction after sperm binding. Next, sperm release lysin from their acrosomal granules, a nonenzymatic protein that dissolves a hole in the vitelline envelope through which the sperm swims. Sperm then contact the egg surface coat, a network of uniformly sized filaments lying directly above the egg plasma membrane. This layer mediates attachment of sperm, via their acrosomal process, to the egg surface. © 1995 Wiley-Liss, Inc.     

An egg envelope component induces the acrosome reaction in sturgeon sperm

The acrosome reaction in Acipenser transmontanus sperm can be induced by a 66,000 dalton glycoprotein that is present in Layer 3 (L3) of the egg envelope and in egg water only following exposure of the eggs to fresh water. When egg water is fractionated on Sepharose CL 6B, the 66,000 dalton glycoprotein-containing fractions possess acrosome reaction inducing activity. Egg water may be species-specific in its ability to elicit the acrosome reaction, as demonstrated by the fact that it has no effect on the sperm of Acipenser fulvescens. Egg jelly possesses no acrosome reaction, inducing activity. The major carbohydrate-containing component of the egg envelope is L3, a layer that contains galactose residues. L3 possesses a 70,000 dalton glycoprotein prior to freshwater exposure and lacks the 66,000 dalton component. If isolated from polyacrylamide gels, the 70,000 dalton glycoprotein elicits acrosome reactions in what appears to be a species specific manner. After freshwater exposure, L3 contains both the 70,000 dalton glycoprotein and the 66,000 dalton glycoprotein that is also present in egg water. The appearance of the 66,000 dalton inducer can be blocked by the incubation of eggs in fresh water containing inhibitors of trypsin activity. Thus, the soluble inducer in egg water may be proteolytically derived from a higher molecular weight complex in the egg envelope.     

Some aspects of water absorption by the developing egg of Schistocerca gregaria

Schistocerca gregaria eggs absorb water over their entire surface. Water entry is facilitated by changes in the permeability of cuticular membranes covering the egg. The shell is markedly more permeable during the period of rapid water uptake and much less so on the other days. The changes in permeability are due to the differing amounts of lipoid material in the shell at different developmental periods. The absorption appears to be an active process requiring oxidative energy, although osmosis may also be partly involved. Water entry is mediated by the serosa and is not influenced by the developing embryo.     

Uptake and loss of water in diapause and non-diapause eggs of crickets

ABSTRACT. In laboratory-laid eggs of the striped ground cricket, Allonemobius fasciatus DeGeer (Gryllidae), water absorption occurs at an early stage of embryogenesis (stage II) at 30°C but is delayed until later stages at lower temperatures. This is related to the variation in diapause stage at different temperatures. No egg developed beyond stage VII (the end of anatrepsis) without water absorption.
A. fasciatus shows seasonal variation in the stage of water absorption. At 30°C, eggs collected in August absorb water at the early stage while many of those collected in September avert diapause and absorb water at a later stage.
Diapause also influenced the water absorption of eggs in A. socius Scudder. Eggs of short-day females enter diapause and absorb water at stage II, while those of long-day females develop without diapause and absorb water at a later stage (around stage IV).
The susceptibility to desiccation (r.h. 50%) was examined at 20°C with A. fasciatus eggs. The percentage water loss and mortality of eggs varied with the time and duration of exposure to desiccation. Eggs are most sensitive to desiccation during the first several days after being laid and during the period of water absorption.     

Gamete interaction in the white sturgeon Acipenser transmontanus: a morphological and physiological review

Synopsis Sturgeon gametes differ from those of most fish in that the sperm possess acrosomes that undergo exocytosis and filament formation while the eggs possess numerous micropyles. Acipenser transmontanus eggs are encased by multilayered envelopes that consist of outer adhesive jelly coats and three structured layers interior to the jelly. The glycoprotein jelly layer only becomes adhesive upon exposure to freshwater. The layer interior to the jelly, layer 3, is the other carbohydrate-containing component of the egg envelope. This layer consists of a water-insoluble glycoprotein that, upon freshwater exposure, is hydrolyzed by a trypsinlike protease to yield a water-soluble, lower molecular weight carbohydrate-containing component. This component can be identified in the surrounding medium when unfertilized eggs are incubated in freshwater. This egg water component elicits acrosome reactions only in homologous sperm. The A. transmontanus sperm acrosome reaction is a Ca⁺⁺ and/or Mg⁺⁺ dependent event that includes the formation of a 10 μ long fertilization filament. A. transmontanus fertilization can occur at low sperm per egg ratios; however, crossfertilization of A. transmontanus eggs with lake sturgeon, A. fluvescens, sperm results in a very low number of fertilized eggs, even at high sperm per egg ratios. The morphological, physiological, and biochemical phenomenon reviewed in this paper are related to the environment in which they occur. Also, the possible role of the acrosome and the presence of numerous micropyles are discussed.     

Cortex reorganization of <Emphasis Type="Italic">Xenopus laevis</Emphasis>eggs in strong static magnetic fields

Observations of magnetic field effects on biological systems have often been contradictory. For amphibian eggs, a review of the available literature suggests that part of the discrepancies might be resolved by considering a previously neglected parameter for morphological alterations induced by magnetic fields – the jelly layers that normally surround the egg and are often removed in laboratory studies for easier cell handling. To experimentally test this hypothesis, we observed the morphology of fertilizable Xenopus laevis eggs with and without jelly coat that were subjected to static magnetic fields of up to 9.4 T for different periods of time. A complex reorganization of cortical pigmentation was found in dejellied eggs as a function of the magnetic field and the field exposure time. Initial pigment rearrangements could be observed at about 0.5 T, and less than 3 T are required for the effects to fully develop within two hours. No effect was observed when the jelly layers of the eggs were left intact. These results suggest that the action of magnetic fields might involve cortical pigments or associated cytoskeletal structures normally held in place by the jelly layers and that the presence of the jelly layer should indeed be included in further studies of magnetic field effects in this system.     

The jelly canal marker of polarity for sea urchin oocytes, eggs, and embryos

The jelly coat of a sea urchin egg possesses a narrow conical channel which identifies the animal pole. This rarely seen structure was first reported by Boveri in 1901 and is easily demonstrated by immersing freshly ovulated eggs into an ink suspension. The jelly canal, as this feature is called, fills with ink particles as the jelly swells. The jelly canal occurs on full-sized primary oocytes and unfertilized eggs. When filled with ink it serves as a useful marker of the animal pole during fertilization and early development when it is not otherwise visible. A common synonym for the jelly canal is ‘micropyle’, but this is a misnomer because sperm do not selectively pass through it for fertilization. The presence of the jelly canal on oocytes suggests how it might form and does not prove that the animal-vegetal polar axis in sea urchin eggs is defined before meiotic maturation.     

Xenopus laevis egg jelly contains small proteins that are essential to fertilization.

The eggs of Xenopus laevis are surrounded by investment layers of egg jelly that interact with the sperm immediately prior to fertilization. Components of these egg jelly layers are necessary for the fertilization of the egg by incoming sperm. Eggs which are stripped of their jelly layers are refractile to fertilization by sperm, but the addition of solubilized jelly promotes fertilization. We have shown previously that the egg jelly layers are composed of a fibrous network of glycoconjugates which loosely hold smaller diffusible components. Extracts of these diffusible components were prepared by incubation of freshly ovulated eggs in high-salt buffers for 12 h at 4 degrees C. This diffusible component extract, when incubated with sperm, promoted the sperm's ability to fertilize dejellied eggs in a dose-dependent manner. In contrast, the high-molecular-weight "structural" glycoconjugates of jelly that remain after extraction of the diffusible components did not increase fertilization efficiency of dejellied eggs nor did nonspecific proteins, carbohydrate polymers, or organic polymers. The diffusible components, analyzed by SDS-PAGE, consisted of a mixture of proteins from 4 to 180 kDa. The protein responsible for fertilization rescue appeared to be <50 kDa and appeared to self-aggregate or to bind to larger proteins. This protein component was required during sperm binding to the egg, its action required an intact egg vitelline envelope, and its action was independent of large soluble polymers such as Ficoll.     

怀头鲇胚胎发育过程卵膜形态结构变化及对孵化影响

采用扫描电镜和光学解剖镜,对黑龙江水域怀头鲇(Silirus soldatovi)成熟卵膜层次构造和受精卵胚胎发育过程中卵膜形态结构变化进行观察,并比较未脱黏和人工脱黏卵受精卵膜的表面超微结构变化.结果显示,受精卵膜的胶膜表面由一层薄而致密的物质组成,上有微孔构造.未脱黏受精卵膜表面胶膜光滑致密,多孔隙,内有小梁相连,随胚胎发育逐渐膨胀、展开、变薄,破膜期自然脱落.人工脱黏几乎全部脱去鱼卵的胶膜层,从而使卵失去黏性.脱去胶膜层的受精卵膜表面由不规则的颗粒状结构紧密嵌合而成,表面粗糙,胚胎发育过程中颗粒形状变化不大,但颗粒层逐渐变薄而且疏松,直至胚胎破膜而出:胚胎发育后期颗粒层有过早脱落和破洞出现.同时对活体鱼卵进行连续比较观察,讨论了卵膜结构及动态变化与孵化效果的关系.     

When do sperm of the sea urchin,Pseudocentrotus depressus,undergo the acrosome reaction at fertilization?

Jelly coats of the sea urchin, Pseudocentrotus depressus, were stripped off the eggs, and the eggs were “inseminated.” After penetration through the isolated jelly hull, sperm swarmed in the cavity previously occupied by the egg. Electron microscopic examination could not detect any sperm with reacted acrosome. Observation was also made of the sperm penetrating through the intact jelly coat-egg complex. Although a number of sperm were examined in ultrathin sections, only those attached to the vitelline layer had undergone the acrosome reaction; those sperm embedded in jelly but not attached to the vitelline layer had not undergone the acrosome reaction. The sequence of events in fertilization of this species and of other echinoids is discussed.     

Alteration of structure and penetrability of the vitelline envelope after passage of eggs from coelom to oviduct in Xenopus laevis.

The vitelline envelope (VE) that surrounds an egg released from the ovary into the coelom of Xenopus laevis differs markedly, in structure and penetrability, from the VE surrounding an oviposited egg. In a coelomic egg, the filaments that form the VE are arranged in distinct fascicles or bundles. The exterior surface of the VE is irregular in contour and is permeated by channels. In an oviposited egg, the filaments are evenly dispersed and lack a fasciculated arrangement; the exterior surface is smooth and no channels are present. The fascicular arrangement of fibrils in the coelomic VE is maintained only at neutral pH, and is not visibly altered by the cortical reaction. VEs from coelomic eggs retain their fasciculated morphology after isolation from the egg. In an in vitro test system, sperm penetrated VEs isolated from oviposited eggs, but failed to penetrate VEs isolated from coelomic eggs. The structural transformation of the VE from the coelomic type to the oviposited type occurs in the first 1-cm segment of the oviduct, prior to addition of jelly to the egg. Neither intact jelly, solubilized jelly, nor jelly extracts were capable of altering the structural organization of coelomic VEs, suggesting that the structural transformation of the VE is effected by some oviducal factor other than jelly.     

Egg envelope synthesis and chorion modification after oviposition in the dragonfly Libellula depressa (Odonata, Libellulidae).

Libellula depressa (Odonata, Libellulidae) is an exophytic dragonfly ovidepositing eggs in clutches on the surface of floating plants and algae. The present work investigates, at ultrastructural level, the gradual differentiation of the egg envelopes and the chorionic changes after egg deposition in water. The ovary of the mature female of L. depressa is composed of numerous strings of panoistic ovarioles, where the eggshell formation takes place gradually throughout the activity of the follicle cells. The present data show that the egg envelopes are constituted of a very thick electrondense vitelline envelope, a thin endochorion and an extremely thick exochorion composed of a fibrillar matrix resting on a thin electrondense layer. After deposition in water, L. depressa eggs, initially white and almost transparent, gradually become brown spots in a semitransparent jelly coat, rich of incorporated debris. The jelly coat enveloping the eggs of L. depressa derives exclusively from the exochorion, constituted of a fibrillar matrix, which swell at contact with water. The jelly-like coat performs an adhesive function and presumably a protective role during egg segmentation and ensuing larval hatching.     

Induction of cross-fertilization between sea urchin eggs and starfish sperm by polyethylene glycol treatment

Cross-fertilization between sea urchin eggs (Strongylocentrotus nudus) and starfish sperm (Asterina pectinifera) was induced by treatment with polyethylene glycol (PEG). Without treatment with PEG, the denuded egg surface (jelly coat- and vitelline coat-free) engulfed the head of acrosome-reacted sperm; however, sperm penetration did not occur [Kyozuka and Osanai, 1988]. When these eggs were exposed briefly to PEG (molecular weight 3,000) in seawater, the sperm entered the egg by membrane fusion. Cortical granules were discharged, and embryogenesis began following sperm penetration. PEG did not induce parthenogenesis in Strongylocentrotus eggs. Egg activation is thus closely linked with gamete membrane fusion.