Temperature‐mediated survival,development and hatching variation of Pacific cod Gadus macrocephalus eggs

Laboratory‐validated data on the survival, development and hatching responses of fertilized Pacific cod Gadus macrocephalus eggs from the northern Japan stock were determined through an incubation experiment. The optimum temperature for survival until hatching ranged from 4 to 8° C. No significant difference in development rates was found between the populations from Mutsu Bay, Japan, and western Canadian coastal waters even though the samples may belong to different G. macrocephalus stocks. Gadus macrocephalus larvae hatched asynchronously from egg batches despite incubation under the same environment during their development. Both incubation temperature and temperature‐mediated hatch rank affect size and yolk reserve. These data suggest that variations in water temperatures within an ecological range markedly influence the development rates, survival and hatching of the eggs, as well as the stage at hatch larvae of G. macrocephalus. Asynchronous hatching and the production of offspring with variable sizes and yolk reserves are considered evolutionary bet‐hedging strategies that enable the species to maximize their likelihood of survival in an environment with variable temperatures.     

Morphology of unfertilized mature and fertilized developing marine pelagic eggs in four types of multiple spawning flounders

Starry flounder Platichthys stellatus, spotted halibut Verasper variegates, turbot Scophthalmus maximus, and Japanese flounder Paralichthys olivaceus are four commercially cultivated multiple spawning flounders that spawn pelagic eggs. Through appropriate light and scanning electron microscope processing, the shape and surface structures (such as micropyle, pores, pore density, and paten) of unfertilized mature and fertilized developing eggs of the four species were observed and measured. First, individual or intraspecific comparisons of the surface structures of eggs at different developmental stages were made. Second, interspecific differences among the four species at the same developmental stage of unfertilized mature eggs were statistically computed and analyzed through one-way analysis of variance and hierarchical cluster analysis. Eggs of the same species collected at different stages of development tend to be different in morphology. Smoothing of the convoluted egg envelope surface and closure of the micropyle to serve as a final step of the polyspermy-preventing reaction are common after fertilization. Based on detailed morphology of micropyle of just-mature fertilizable eggs, turbot, starry flounder, and Japanese flounder each have a micropyle with a long canal but no distinct micropylar vestibule, type III of Riehl and Götting (Arch Hydrobiol 74:393–402, 1974). In contrast, spotted halibut has a micropyle with a distinct flat micropylar vestibule and a long canal, type II. Envelope surface microstructures, especially those in the micropyle region, are useful characters for egg identification among the four species. Cluster analysis using selected egg characters indicated the highest similarity between turbot and Japanese flounder and that starry flounder is obviously more similar to turbot and Japanese flounder than to spotted halibut.     

The surface structure of Antarctic fish eggs and its use in identifying fish eggs from the Southern Ocean

Summary The surface structure of eggs of Notothenia neglecta, Nototheniops larseni, Nototheniops nudifrons (Nototheniidae), Champsocephalus gunnari and Chaenocephalus aceratus (Channichthyidae), collected near Elephant Island (Atlantic sector of the Southern Ocean) and of Notothenia rossii rossii (Nototheniidae) from Kerguelen waters (Indian sector of the Southern Ocean) was studied by scanning electron microscopy (SEM). Eggs of N. larseni differed from the other species by containing a micropyle with two micropyle pits. External morphological characteristics of the egg envelope such as the ultrastructure of the zona radiata, its interpore distances and the diameter of the micropyle were species-specific and could aid in the identification of fish eggs collected from the Southern Ocean.     

Closure of the micropyle during embryonic development of some pelagic fish eggs

The fine structure of the egg envelope and micropyle was studied in unfertilized and developing eggs of the flounder Paralichthys olivaceus (Temminck & Schlegel), the Alaska pollack Theragra chalcogramma (Pallas), the Japanese tilefish Branchiostegus japonicus (Houttuyn) and the porgy Pagrus major (Temminck & Schlegel). The outer envelope surface of the unfertilized egg was wrinkled, while the inner surface was folded. The micropyle of the unfertilized egg consisted of a shallow vestibule and a distinct canal. The micropylar region of the inner surface of the envelope had a conical- or bowl-shaped protrusion. In developing eggs, the thickness of the envelope decreased and showed smooth outer and inner surfaces which indicated that it had been stretched tangentially at the time of the perivitelline space formation. The lumen of the micropylar canal was invariably occupied with envelope material. We postulate that the blockage of the micropylar canal is a result of the stretching of the envelope. The closure of the micropyle inhibits sperm and external pathogens from penetrating into the perivitelline space and seems to be involved in both the permanent prevention of polyspermy and the protection of the developing embryo from bacterial infection.     

Experimental Parameterisation of Principal Physics in Buoyancy Variations of Marine Teleost Eggs

It is generally accepted that the high buoyancy of pelagic marine eggs is due to substantial influx of water across the cell membrane just before ovulation. Here we further develop the theoretical basis by applying laboratory observations of the various components of the fertilized egg in first-principle equations for egg specific gravity (ρ_egg) followed by statistical validation. We selected Atlantic cod as a model animal due to the affluent amount of literature on this species, but also undertook additional dedicated experimental works. We found that specific gravity of yolk plus embryo is central in influencing ρ_egg and thereby the buoyancy. However, our established framework documents the effect on ρ_egg of the initial deposition of the heavy chorion material in the gonad prior to spawning. Thereafter, we describe the temporal changes in ρ_egg during incubation: Generally, the eggs showed a slight rise in ρ_egg from fertilization to mid-gastrulation followed by a gradual decrease until full development of main embryonic organs just before hatching. Ontogenetic changes in ρ_egg were significantly associated with volume and mass changes of yolk plus embryo. The initial ρ_egg at fertilization appeared significantly influenced by the chorion volume fraction which is determined by the combination of the final chorion volume of the oocyte and of the degree of swelling (hydrolyzation) prior to spawning. The outlined principles and algorithms are universal in nature and should therefore be applicable to fish eggs in general.     

Sperm Penetration and in vitro Fertilization of the Egg of the House Cricket Acheta domesticus

Summary

A study of sperm penetration of the egg of the house cricket, Acheta domesticus, using a fluorescent microscope technique, showed that sperm penetration of the chorion, prior to fertilization, is not restricted to a specialised area of the egg surface, such as the micropyle. An acrosomal filament is seen on penetrating sperm. Polyspermy (multiple sperm attachment) is seen under normal conditions.

Eggs fertilized in vitro developed to the 4 day (pre-katatrepsis) stage, but did not undergo katatrepsis. Development was confirmed by cytogenetic studies. The percentage of eggs showing cleavage nuclei (i.e. initial development) was 59% after in vitro fertilization and 5% in ovarian eggs incubated in a hypotonic medium.     

The orientation of the dorsal/ventral axis of zebrafish is influenced by gravitation

Following fertilization of eggs of Brachydanio rerio a blastodisc is formed which, by cleavage divisions and epibolic movements, gives rise to a cell cap covering the yolk. When about half of the yolk is covered by the cells, a thickening appears at the progressing margin, the embryonic shield, which gives rise to the axial organs of the future embryo. Thereby, the dorsal/ventral axis can be recognized. Our experiments show that the blastodisc is formed at the position of the micropyle, the only site where the sperm can enter the egg and oocyte. Later the head appears at this position. We show that the position of the embryonic shield on the circumference of the cell cap is influenced by gravitation. We also demonstrate that there is no correlation between the orientation of the first or second cleavage plane and the position of the embryonic shield. Correspondence to: K. Herrmann     

Staging and the time course of embryonic development in kurosoi,Sebastes schlegeli

Synopsis Intraovarian development of embryos was observed in 31 female rockfish kurosoi,Sebastes schlegeli, which had been raised in the laboratory. Three of the females had newly fertilized eggs; the others' embryos were at various stages. The developmental series (33 stages) was not very different from that in oviparous fishes. However, several characteristic features were noted: thin egg envelope, narrow perivitelline space, elliptical encapsulated embryo in the late stages and depletion of yolk at parturition. The time course of development up to birth at different temperatures was adjusted to that at 9.8°C using the Arrhenius equation and processed for the relationship between the stage number (X) and stage-to-birth time in days (Y). An equation, was obtained, and the gestation period was calculated to be 48 days at 9.8°C.     

Diel vertical migration of adult Pacific cod Gadus macrocephalus in Alaska

The diel vertical migration (DVM) of Pacific cod Gadus macrocephalus was examined using depth and temperature data from 250 recaptured archival tags deployed on G. macrocephalus in the eastern Bering Sea and in the Gulf of Alaska near Kodiak Island. DVM of two types, deeper during daytime (type I) and deeper during night‐time (type II), occurred frequently (15–40% of all days) in G. macrocephalus released at all sites. Most individuals displayed both diel types, with each type of behaviour lasting up to 58 contiguous days, and day and night depth differences averaging c. 8 m. Despite high among‐individual variability, the occurrence of DVM varied significantly with the release site, season (i.e. day‐of‐year) and bottom depth, with the trend in seasonal occurrence nearly opposite for type I compared to type II DVM. No significance could be attributed to G. macrocephalus fork length, sex or ambient (tag) temperature. Trends in the magnitude of G. macrocephalus depth change were observed, with increased movement often occurring during night‐time, dawn and dusk, and at release sites where the bathymetry was more complex. Both type I and type II DVMs were attributed to foraging on prey species that also undergo DVM, and increased vertical movements of G. macrocephalus during crepuscular and night‐time periods were attributed to more active foraging during dim‐light conditions when G. macrocephalus can potentially exploit a sensory advantage over some of their prey.     

A 4-year summary of the nonsurgical recovery of baboon embryos: A report on 498 eggs

A nonsurgical embryo recovery procedure, developed to allow the economical acquisition of cleavage stage baboon embryos, has been successfully used for 4 years. With this technique, 498 eggs have been recovered from 979 uterine flushes (50.9%) on 71 baboons. Of 467 eggs recovered from mated baboons, 290 (62.1%) were fertilized. Papio anubis females provided a higher percentage of fertilized eggs (75.3%) than did Papio hamadryas (47.8%) or Papio cynocephalus (44.3%) females following exposure to males during estrus, although sexual preference may be responsible for the reduced fertilization rate in the P. cynocephalus females. Recovery rates from individual baboons ranged from 0% (n = 11) to between 66% and 93% for ten baboons from each of which 12–33 eggs have been recovered. Fertilized eggs were at the two-cell (n = 23) to blastocyst (n = 53) stage at recovery 1–6 days postdeturgescence (PD) of the sex skin, with morulae (n = 84) being the most frequent cell stage recovered (30%). The optimum time for performing the procedure was the third day PD, when 113 (40%) embryos were recovered. The abilities of baboons to become pregnant and to provide fertilized embryos were significantly related (P < 0.005), allowing the embryo recovery technique to be used as a screening procedure for evaluating baboon fertility.     

The embryo pancreas is a source of increased yolk amylase in the fertilized eggs of domestic fowls.

The amylases were studied in the yolk of fertilized eggs and in the pancreases of the embryos of domestic fowls. The amylase activity in the yolk increased markedly from 13 days of incubation until hatching, but the activity decreased when the embryos were taken out of the eggs. The isoamylases in the yolk and in the pancreas of the embryo were identical electrophoretically. The amylase occurs mainly in the pancreas of the embryo. We think that the increase in amylase activity in the yolk of fertilized eggs during incubation depends upon the accumulation of pancreatic amylase synthesized by the developing embryo in the egg.     

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.     

Scanning electron microscopy of the post-embryonic stages of the climbing perch,Anabas testudineus

Different developmental stages, fertilized eggs through hatchlings, of the climbing perch,Anabas testudineus, have been studied by scanning electron microscopy. The surface specialization of eggs and hatchlings reveals that while the egg surface is reticulate in appearance, the hatchlings are covered with microridges. Vitelline arteries are seen at the pharyngeal and abdominal regions. They supply nutrients directly from the yolk sac to the developing embryo. Three pairs of such arteries are distinctly seen in the pharyngeal region. Mucous glands are discernible at places over the entire body surface of the embryo before the formation of scales. The skin seems to be helpful in gaseous exchange till the gills and accessory respiratory organs develop and become functional.     

Fine structures of the micropyles of pelagic eggs of some marine fishes

The micropyles and their surrounding structures of pelagic fish eggs of nine species were examined and compared using a scanning electron microscope (SEM). The surface of the egg membrane possesses uniformly distributed pores. However,Lateolabrax japonicus possesses a number of small knobs. The region of the micropyle of fish eggs examined is characterized by a circular elevation in which the pores are larger than those on the other parts of the egg membrane. In contrast,Kareius bicoloratus egg has no elevation and possesses uniform pores to the edge of the micropylar canal. The funnel-like structure is seen in the opening of the micropyle inSardinops melanostictus, Saurida elongata, Sillago japonica, Parapristipoma trilineatum, Hypodytes rubripinnis andK. bicoloratus. These specific features of the micropyle may be used for the identification of pelagic eggs.     

Early embryonic development,including germ-disk stage,in the theridiid spider Achaearanea japonica (Bös. et Str.)

Early embryonic development, from the first cleavage to the germ-disk stage, in the theridiid spider Achaearanea japonica was examined by light and electron microscopy. The eggs are syncytial during the first four cleavages, and then invaginations of cell membranes fuse to generate the blastomeres at the sixteen-nucleus stage. The cleavage pattern is a modified type of total cleavage. It appears that radial bundles of microtubules that radiate from the perinuclear cytoplasm may participate in the migration of cleavage nuclei for the formation of the blastoderm. The large yolk granules are sequestered by cell membranes from the blastomeres or blastoderm cells into the interior of the embryo together with various organelles and glycogen granules. Most of the blastoderm cells converge in the upper hemisphere to form the germ disk, whereas a few cells remain in the lower hemisphere. The embryo at the germ-disk stage contains many spherical germ-disk cells. Almost no large yolk granules are found in these cells, but the flat remaining cells each contain several large yolk granules. These remaining cells may preserve a flat shape to cover the surface of the embryo that does not include the germ disk. © 1995 Wiley-Liss, Inc.     

Proteolytic processing of Blattella germanica vitellin during early embryo development

In the eggs of the cockroach Blattella germanica, vitellin (Vt) utilization is initiated 4 days postovulation by the proteolytic processing of its three subunits. These reactions yield a specific set of peptides that are consumed by the developing embryo. A yolk proteinase activity, believed central to this processing event, has been investigated. First expressed at day 3 postovulation, just prior to Vt's processing, its specific activity with synthetic substrates increased four-fold to 18-fold through day 6. In addition, a mixing experiment showed that these proteinases(s) can also process Vt's large subunits in vitro. A relationship between Vt processing and proteinase specific activity was also noted with two B. germanica translocation heterozygotes, which displayed differences in the extent of Vt processing. One group of eggs (group A) failed to process any Vt subunit. A second group (B) processed the M_r 102,000 subunit but not the M_r 95,000. A third group (C) processed their Vt normally. Proteinase specific activities in the yolk of translocant's eggs at day 6 mirrored the extent of processing, being highest in group C eggs and effectively absent from the yolk of group A eggs. Eggs defective in Vt processing also contained arrested embryos. It is concluded that the yolk proteinase activity described here participates in Vt processing at day 4 postovulation. Microscopic examination of yolk obtained from eggs of wild type females showed that, as processing began in vivo (day 4), the yolk granules also underwent an abrupt decrease in size from diameters of 15–30 μm to 3–10 μm. Yolk granules of those translocant's eggs that were defective in Vt processing did not undergo this size decrease, suggesting that granule reorganization and Vt proteolysis may be linked functionally.     

The use of poly-L-lysine to facilitate examination of sperm entry into pelagic, non-adhesive fish eggs

The fish egg is surrounded by a thick envelope called the chorion. The fertilizing spermatozoon enters the egg through a canal-like structure in the chorion, the micropyle. Examination of micropyle at fertilization is difficult if eggs are large and have no distinct landmarks surrounding the micropyle, or if they are positively buoyant in water. Eggs of many commercially important fishes (e.g., flounder, sea bream and eel) are buoyant in water or only slightly adhere to solid objects (e.g., sands, rock and water plants), which makes observation of spermatozoa at fertilization difficult. Here, we report that such eggs can be firmly attached to plastic and glass dishes that have been previously coated with poly-L-lysine. These adhering eggs can be fertilized and develop normally on the dishes. Observations of micropyles of three fish species, before and after sperm entry are presented.