Laboratory spawning of the purple snail Plicopurpura pansa (Gastropoda: Muricidae)

The spawning of the muricid gastropod Plicopurpura pansa in the laboratory at 22-23 degrees C is described. Females deposited 1-20 capsules daily for at least 20 weeks, and produced up to 150 capsules each per spawning season. During spawning, egg clusters were formed consisting of hundreds of capsules of different ages deposited by different females. Each egg capsule contained an average of 436 embryos (+/- s.d. 213.6, range: 95-1092, n=50). Embryos developed without nurse eggs. After six to eight weeks of intracapsular, lecithotrophic development, planktotrophic veligers hatched with two fully developed velar lobes.     

Ability of some gastropod egg capsules to protect against low-salinity stress

The ability of the egg capsules of three intertidal gastropod species to protect embryos against low-salinity stress was examined. Encapsulated embryos of Ilyanassa obsoleta (Say), Nucella lamellosa (Gmelin), and N. lima (Gmelin) were far more tolerant of transfer to water of reduced salinity than were embryos which had been prematurely removed from egg capsules and transferred to low-salinity sea water directly. However, the walls of N. lamellosa and N. lima capsules were found to be permeable to salts and at least to small carbohydrate molecules (glucose). Correspondingly, indirect evidence for all three species and direct evidence for N. lamellosa indicates that the osmotic concentration of intracapsular fluid declines to near ambient after transfer of egg capsules to dilute medium. Experiments conducted using embryos of N. lamellosa suggest that the egg capsules may protect embryos by reducing the rate at which the osmotic concentration of intracapsular fluid decreases rather than by reducing the magnitude of the decrease.     

Embryonic development and reproductive seasonality of Buccinanops globulosus (Nassariidae) (Kiener, 1834) in Patagonia,Argentina

Buccinanops globulosus mated all year round, with higher frequency from May to September, prior to spawning months. Gravid females were found between October and March. Oviposition peaked during rising temperatures and longest daylength while hatching peaked with high water temperature and declining daylength. Gravid females measured between 20 and 41?mm in shell length. The spawn consisted on average of 31 egg capsules, each containing 1266 eggs. Embryos usually completed development within each egg capsule by ingesting small fragments of the uncleaved nurse eggs, which were not a limiting resource. Egg capsules with more than one embryo were not common; in those cases, the embryos had different sizes probably related to intracapsular competition for nutrients, and were on average smaller than solitary embryos in the other capsules. Embryos hatched as crawling juveniles with a mean hatchling shell length of ~3.4?mm. In a few cases, malformed embryos were found, but it was not a common phenomenon. The information recorded in this study, as the minimum reproductive size and spawning season, is valuable for fisheries management.     

Capsule walls as barriers to oxygen availability: Implications for the development of brooded embryos by the estuarine gastropod Crepipatella dilatata (Calyptraeidae)

Encapsulation of developing embryos imposes potential restrictions, because the capsule wall must allow for adequate inward diffusion of oxygen and for increased diffusion of oxygen as metabolic demand increases with continued development. Samples of egg capsules from the gastropod Crepipatella dilatata were used to document surface characteristics, composition of the different capsule wall layers, and alterations in wall thickness during development. The diffusion coefficient and capsule wall permeability were determined experimentally for capsules containing embryos at different developmental stages. We also determined oxygen consumption rates for various embryonic stages and for nurse eggs, which provide food for embryos during development. The capsule wall of C. dilatata possesses 2 differentiated layers: the external capsular wall (ECW) and the internal capsular wall (ICW). The ECW is compact and fibrous, features that remain invariable during development, and lacks surface features that might make some portions of the capsule wall more permeable to oxygen than others. On the other hand, the ICW is initially spongy and thick, but significantly decreases in thickness over time, particularly before the embryos begin feeding on nurse eggs. Although the capsule wall is a serious barrier to diffusion, permeability to oxygen increases over time by 112% due to the dramatic thinning of the inner capsule wall layer. Nurse eggs consume oxygen but at very low rates, supporting the idea that they correspond to living embryonic cells that have stopped their development. Respiration measurements indicated that embryos are initially supplied with enough oxygen within the egg capsules to carry out the activities characteristic of embryogenesis, even though the capsular walls show their maximum thickness and lowest permeability at this time. However, as the embryo develops its velum and becomes more active, capsule wall thickness decreases and capsule permeability to oxygen increases. Correspondingly, the oxygen demands of metamorphosed but still encapsulated specimens are approximately 135% higher than those of pre-metamorphosed sibling embryos.     

Early development of the limpet Siphonaria lessonii Blainville, 1827 in populations affected by different physical stressors

The limpet Siphonaria lessonii is very common along Atlantic Patagonian intertidal rocky shores. We studied the early intracapsular embryonic development of this limpet in detail in two populations in north Patagonia, with different environmental conditions (i.e. wave exposure, wind, temperature). Early development in both populations was achieved at controlled and equal conditions (13°C). The spawn consisted of a series of enchained egg capsules embedded in a jelly mass. The development from egg to hatching veliger took 9–11 days in embryos from both populations. The developmental process at both sites was identical, differing only in the embryos' sizes at each stage. Larger adult individuals producing larger embryos were registered at the sheltered site. The differences in sizes of adult and embryos of S. lessonii could be attributed to distinct environmental stressful conditions between sites.     

Costs of parental care in the gastropod Conus pennaceus: Age-specific changes and physical constraints

Summary Age-specific changes in the allocation of reproductive energy to protective capsules, ova and intracapsular fluid are documented for the marine gastropod Conus pennaceus. As female snails grow in shell length they produce larger capsules with thicker and stronger walls. Because large capsules contain lower densities of ova than do small ones, growing females must increase the number, as well as the size, of egg capsules they produce. As a result of this pattern of ova packaging, per ovum costs of encapsulation (parental care) increase with increasing female size and age. The data suggest that the number of embryos a capusle can support may be limited by respiratory constraints related to capsule surface area or wall thickness. As capsule size increases, surface/volume ratios decline and capsule wall thickness increases. Either of these processes should result in a reduction in net gas transport per unit of capsule contents.     

ENCAPSULATION OF EGGS BY MARINE GASTROPODS: EFFECT OF VARIATION IN CAPSULE FORM ON THE VULNERABILITY OF EMBRYOS TO PREDATION

Representatives of many plant and animal taxa enclose their embryos within some form of protective structure. Inter- and intraspecific differences in the morphology of these egg coverings may have profound effects on the development and survival of encapsulated embryos, yet in many taxa little is known about the causes or potential consequences of this variation. Comparisons of capsule morphology among populations of the rocky shore gastropod, Nucella emarginata, revealed significant variation in the thickness of capsule walls, the only barrier separating developing embryos from the external environment. Laboratory experiments demonstrated that thicker-walled capsules were more resistant to predation by a co-occurring isopod, Gnorimosphaeroma oregonense, than were thinner-walled capsules. Control experiments confirmed that these differences in vulnerability were not caused by differences in the palatability of the capsule wall or attractiveness of the capsule contents. The actual mechanism by which thick-walled capsules differentially protect developing embryos remains unclear, although decreased vulnerability of thick-walled capsules to these isopods may simply result from increased handling time by predators. Subtle differences in capsule morphology thus appear to have substantial effects on the survival of encapsulated embryos. Hence, predators may have played an important role in selecting for the production of thick-walled capsules among populations of N. emarginata.     

Ion and Osmoregulation in Prenatal Elasmobranchs: Evolutionary Implications

The elasmobranchs represent a fascinating series of experimentsin the evolution of maternal support for developing embryos.In oviparous species, eggs are enclosed in a tough, fibrouscapsule. The capsule is very permeable and the embryonic tissuesare bathed in a solution ionically similar to sea water withinhours of oviposition. In the primitively viviparous speciesSqualus acanthias, early embryos in egg capsules are retainedin utero and are bathed in a solution similar to maternal plasma.Several months into the 22 month gestation period the embryosare capable of independent iono- and osmoregulation in a uterinesolution that resembles sea water. Embryos of more advancedviviparous species develop in a solution that is similar tomaternal plasma. Iono- and osmoregulation by these embryos wouldappear to beminimal. It is clear that in the oviparous elasmobranchs,the ability of the egg/embryo to maintain salts and urea atappropriate levels is present at the earliest stage of development.The ability of prenatal elasmobranch embryos to iono- and osmoregulatewould allow the evolution ofa diverse array of reproductivestrategies in the elasmobranchs.     

Development and the influence of nurse egg allotment on hatching size in Searlesia dira (Reeve, 1846) (Prosobranchia : Buccinidae)

The reproductive biology of the intertidal prosobranch Searlesia dira (Reeve, 1846) was examined with special attention given to variability in the nurse egg to embryo ratio among capsules, among clutches and among geographically isolated populations. Embryos and nurse eggs were distributed among the capsules in a manner consistent with the hypothesis that nurse eggs were genetically predetermined, that each female had a genetically defined nurse egg to embryo ratio, and that each capsule represented a random sample of that ratio. The binomial distribution of embryos and nurse eggs among the capsules resulted in some capsules receiving many more embryos per nurse egg than others. The number of nurse eggs an embryo succeeded in eating was proportional to the number of capsule-mates sharing a capsule. Embryos eating more nurse eggs hatched out at a larger size. Differences in the nurse egg to embryo ratios among capsules in the same clutch were much larger than that of the mean ratios among clutches. Among-site differences in the mean nurse egg to embryo ratios suggest that selection pressure for different mean hatching sizes may have acted on the mean nurse egg to embryo ratios.In contrast to the predictions of optimal hatching size theory, hatching size varied widely within clutches as a consequence of differences in nurse egg to embryo ratios among capsules. This variance may be adaptive for species that lay their eggs months before juveniles emerge into an unpredictable environment, or simply be a consequence of an imperfect mechanism for increasing hatching size.     

Embryonic development of the false limpet Siphonaria lateralis from Atlantic Patagonia

Siphonariids are pulmonate gastropods inhabiting rocky intertidal habitats, and many studies have focused on these false limpets around the world. In the southern South Atlantic, studies on reproduction and development in species of Siphonaria are scarce. We studied the embryonic development and egg masses of Siphonaria lateralis at its northernmost distribution in Atlantic Patagonia. In S. lateralis, as in most species of Siphonaria, individuals spawn benthic egg masses that strongly attach to intertidal rocky substrata. A single spherical egg that measures ~120 µm develops inside the egg capsule of S. lateralis. Considering the relatively small egg size, and reports from previous studies, the developmental modality of S. lateralis might be expected to include a planktotrophic larval phase. However, we found that hatchlings emerged as 1‐mm crawling juveniles, probably owing to the presence of intracapsular fluid, which may provide the energetic requirements for direct development. The embryonic size changed little from the egg to veliger stages, and then increased rapidly until the hatchling stage. We compared development in S. lateralis with development in the sympatric Siphonaria lessonii, in which egg size was reported to be ~80 µm and hatching occurs as planktotrophic veliger larvae. In these two species, spawn and early intracapsular developmental modes are remarkably different; these differences represent contrasting ways to survive in the harsh and physically stressful intertidal Patagonian coasts.     

Functional morphology of embryonic development in the Port Jackson shark Heterodontus portusjacksoni (Meyer)

The oviparous Port Jackson shark Heterodontus portusjacksoni embryo has a long incubation of 10–11 months during which it undergoes major morphological changes. Initially the egg capsule is sealed from the external environment by mucous plugs in either end of the capsule. Four months into incubation, the egg capsule opens to the surrounding sea water. Fifteen stages of development are defined for this species, the first 10 occur within the sealed capsule, the remaining five after capsule opening to hatching. The functional significance of major definitive characters such as circulation within the yolk membrane and embryo, rhythmic lateral movement of the embryo, external gill filaments, heart activity, internal yolk supplies, egg jelly and the significance of the opening of the egg capsule are described. The egg jelly in the sealed capsule functions to mechanically protect the embryo during early development, however, it eventually creates a hypoxic environment to the embryo as the available oxygen is used up. This generates several physiological challenges to the developing embryo. It is able to overcome these problems by morphological changes such as increasing the effective surface area for gaseous exchange with the development of external gill filaments, fins and extensive circulation in both the embryo and attached external yolk sac. These adaptations become limiting as the embryo grows and respiratory needs outweigh the available oxygen. At this time, the mucous plugs dissolve and the capsule becomes open to the external environment.     

Effects of in vitro production on horse embryo morphology,cytoskeletal characteristics,and blastocyst capsule formation

Blastocyst formation rates during horse embryo in vitro production (IVP) are disappointing, and embryos that blastulate in culture fail to produce the characteristic and vital glycoprotein capsule. The aim of this study was to evaluate the impact of IVP on horse embryo development and capsule formation. IVP embryos were produced by intracytoplasmic sperm injection of in vitro matured oocytes and either culture in synthetic oviduct fluid (SOF) or temporary transfer to the oviduct of a ewe. Control embryos were flushed from the uterus of mares 6-9 days after ovulation. Embryo morphology was evaluated with light microscopy, and multiphoton scanning confocal microscopy was used to examine the distribution of microfilaments (AlexaFluor-Phalloidin stained) and the rate of apoptosis (cells with fragmented or terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling-positive nuclei). To examine the influence of culture on capsule formation, conceptuses were stained with a monoclonal antibody specific for capsular glycoproteins (OC-1). The blastocyst rate was higher for zygotes transferred to a sheep's oviduct (16%) than for those cultured in SOF (6.3%). Day 7 IVP embryos were small and compact with relatively few cells, little or no blastocoele, and an indistinct inner cell mass. IVP embryos had high percentages of apoptotic cells (10% versus 0.3% for in vivo embryos) and irregularly distributed microfilaments. Although they secreted capsular glycoproteins, the latter did not form a normal capsule but instead permeated into the zona pellucida or remained in patches on the trophectodermal surface. These results demonstrate that the initial layer of capsule is composed of OC-1-reactive glycoproteins and that embryo development ex vivo is retarded and aberrant, with capsule formation failing as a result of failed glycoprotein aggregation.     

Ionic and osmotic environment of developing elasmobranch embryos

Synopsis The elasmobranchs display a variety of ionic and osmotic environments for developing embryos. Oviparous species protect their eggs with a tough, fibrous capsule which is highly permeable to ions and urea even at oviposition. Thus the embryonic tissues are bathed by a solution ionically similar to sea water. In the more advanced reproductive style ofSqualus acanthias (a lecithotrophic live bearer) early embryos in egg capsules are retained in utero and bathed in a solution osmotically similar to maternal plasma. Several months into the 22 month gestation period the embryos can iono- and osmoregulate in a uterine solution resembling sea water. Embryos of more advanced viviparous species develop in a solution that is ionically and osmotically similar to maternal plasma. Iono- and osmoregulation by these embryos would appear to be unnecessary. Clearly, in the oviparous elasmobranchs, the ability of the embryo to regulate salts and urea is present at the earliest stage of development. The need for elasmobranch embryos to regulate osmolytes was reduced or delayed as viviparity evolved.     

Embryonic rotational behaviour in the pond snail Lymnaea stagnalis: influences of environmental oxygen and development stage

Responses of freshwater organisms to environmental oxygen tensions (PO₂) have focused on adult (i.e. late developmental) stages, yet responses of embryonic stages to changes in environmental PO₂ must also have implications for organismal biology. Here we assess how the rotational behaviour of the freshwater snail Lymnaea stagnalis changes during development in response to conditions of hypoxia and hyperoxia. As rotation rate is linked to gas mixing in the fluid surrounding the embryo, we predicted that it would increase under hypoxic conditions but decrease under hyperoxia. Contrary to predictions, early, veliger stage embryos showed no change in their rotation rate under hyperoxia, and later, hippo stage embryos showed only a marginally significant increase in rotation under these conditions. Predictions for hypoxia were broadly supported, however, with both veliger and hippo stages showing a marked hypoxia-related increase in their rotation rates. There were also subtle differences between developmental stages, with hippos responding at PO₂s (50% air saturation) greater than those required to elicit a similar response in veligers (20% air saturation). Differences between developmental stages also occurred on return to normoxic conditions following hypoxia: rotation in veligers returned to pre-exposure levels, whereas there was a virtual cessation in embryos at the hippo stage, likely the result of overstimulation of oxygen sensors driving ciliary movement in later, more developed embryos. Together, these findings suggest that the spinning activity of L. stagnalis embryos varies depending on environmental PO₂s and developmental stage, increasing during hypoxia to mix capsular contents and maintain a diffusive gradient for oxygen entry into the capsule from the external environment (“stir-bar” theory of embryonic rotational behaviour).     

Embryo developmental events and the egg case of the Aleutian skate Bathyraja aleutica (Gilbert) and the Alaska skate Bathyraja parmifera (Bean)

Embryo development events were correlated with egg-case changes for the Aleutian skate Bathyraja aleutica and the Alaska skate Bathyraja parmifera . Yolk absorption underwent two phases: that of steady absorption during early development and that of rapid yolk absorption during the final development stages. Total length ( L _T) for 50% of the pre-hatching embryos egg-case jelly disappearance was 92·04 mm (range 81–102 mm) and 99·36 mm (range 81–100 mm) for B. aleutica and B. parmifera , respectively, allowing the inner chamber to open to seawater flow. The tail filament underwent three phases of growth: rapid elongation during early development (<100 mm embryo L _T), stasis of tail filament length during the remainder of embryo development and rapid absorption soon after hatching. Complete tail filament development coincided with the disappearance of egg-case jelly. Clasper buds first developed at embryos >70 mm L _T for both species and the sex ratio was 1:1 well before hatching. Egg cases that were devoid of an ova or developing embryo were c. 5·0 and 6·5% of the egg cases examined for B. aleutica and B. parmifera , respectively. Measurements showed that egg cases containing only egg jelly were smaller in both width and length than those possessing an ova. Embryo stages were punctuated with distinct events that correlated with egg case changes controlling the internal environment of the developing embryo.     

Reproduction and development in a vermetid gastropod, Vermetus triquetrus

Abstract. We report on a study of reproduction and development in the Mediterranean vermetid gastropod Vermetus triquetrus from the SE coast of Spain. It is a gonochoristic species. The egg capsules are attached to the inside of the shell, and females brood up to 22 capsules simultaneously (more often 4–10). The capsules hold 10–61 eggs or embryos; the uncleaved eggs are yolk-rich, with a mean diameter of 377.3 μm. A distinct polar lobe occurs during the first cleavage, and blastomere D has discernible qualities after the 4-cell stage. The formation of the mesentoblast 4d occurs at the transition from the 24-cell stage to the 25-cell stage. Gastrulation begins after the 36-cell stage. Internal yolk is the major source of nutrition for the encapsulated embryos, but some nurse eggs (∼ 12%) and some sibling larvae are also ingested by the developing embryos. Hatching occurs during the swimming/crawling pediveliger stage, and metamorphosis is completed outside the capsules soon after hatching. Hence, larval development in Vermetus triquetrus is lecithotrophic intracapsular, with a short free-swimming/crawling phase.     

Experimental studies on egg production by Convoluta roscoffensis: Graff, 1882 (Turbellaria,Acoela)

Convoluta roscoffensis collected from the natural habitat in reproductive condition laid egg capsules for up to 16 weeks under laboratory conditions. However, both the number of capsules laid and number of embryos per capsule decreased with time in culture. When animals were maintained in an alternating light and dark regime, the capsules were laid during the dark period. Animals ceased to deposit egg capsules within five days of incubation in the photosynthetic inhibitor, DCMU. This suggests that metabolism of the algal symbionts of C. roscoffensis contributes to egg production.