Organogenesis in normal and lobeless embryos of the marine prosobranch gastropod Ilyanassa obsoleta

The histogenetic and organogenetic capabilities of normal and lobeless embryos of Ilyanassa obsoleta are analyzed. Larvae developed from eggs in which the polar lobe was removed at the trefoil stage of first cleavage were studied in wholemount and section. None showed any evidence of eyes, statocysts, operculum, external shell, heart, or intestine. Sixty per cent of the wholemount larvae observed under polarized light exhibited internal masses of birefringent material resembling shell material. Of eight- to nine-day lobeless larvae studied as stained, sectioned material 100% possessed muscle tissue; 97.5% possessed velar tissue, digestive gland tissue, and style sac tissue; 52.5% possessed two or more style sac areas; 71% exhibited esophagus; 63% possessed stomach tissue; 37% exhibited stomodeal-like invaginations of the ectoderm; 31% exhibited a lightly staining ciliated area at the tip of the prevelar ectoderm; and 21% possessed glandular ectoderm resembling pedal or mantle gland tissue. The evidence presented suggests that the material of the polar lobe has an important role not only in cellular differentiation but also in the organization of tissue types into organs or organ primordia.     

An atlas of light micrographs of normal and lobeless larvae of the marine gastropod Ilyanassa obsoleta

Summary

The olive fruit fly, Dacus oleae, female incorporated dietary ¹⁴ C-labeled malonate, succinate, glutamate, and propionate into the primary pheromone component, 1,7-dioxaspiro[5,5]undecane (1); but did not significantly incorporate acetate. Explanted pheromone glands when incubated in pH 7 phosphate buffer with [¹⁴C]malonate incorporated the label into 1, but did not incorporate [¹⁴C]glutamate. Incorporation of labeled malonate by incubating explanted glands was temperature and age dependent.     

H1 histone subtypes and subtype synthesis switches of normal and delobed embryos of Ilyanassa obsoleta

Histone H1 subtype complexity and H1 histone subtype synthesis switches were characterized during the development of normal embryos of the mud snail Ilyanassa obsoleta. The effect of the removal of the third polar lobe on the normal H1 pattern of synthesis was then investigated in the delobed embryo to determine if classical polar lobe effects are accompanied by a perturbation of these patterns. SDS-gel electrophoresis and fluorography of radiolabeled 5% perchloric acid-soluble nuclear extracts resolved six H1 proteins designated bands 1-6. Bands 1-5 migrate as a cluster of individual bands with similar mobilities. Band 6 has a substantially slower mobility. The synthesis of band 6 is predominant during the first 6 hr post-trefoil. During cleavage and gastrulation bands 1 and 2 are predominant while band 3, 4, and 5 become predominant during organogenesis. In addition, it has been found that removal of the polar lobe delays the off-switch of the early bands 6, 1, and 2 and the on-switch of the late bands 3, 4, and 5. This must result in a different H1 composition in the chromatin of the two embryo types. Cell number data of normal and delobed embryos reveal that the delay in subtype synthesis switching is not caused by an overall delay of cell division in the delobed embryo. However, the data indicate that a subpopulation of cells may not divide, or may divide late, in the delayed embryo. The data also suggest that the D cell lineage may be involved in the control of histone synthesis switching in the A, B, and C cell lineages.     

Trematode accumulation by the estuarine gastropod Ilyanassa obsoleta.

The accumulation of larval trematodes by Ilyanassa obsoleta (Gastropoda) at 2 estuarine sites in Delaware was studied. Initial infection status of snails was assayed by looking for shed cercariae. Native snails (most already infected) were deployed at sites A and B, and sentinel snails (putatively uninfected) were deployed at site B. All were individually marked and, if found, reassessed for infection after being free 1-3 summers. Himasthla quissetensis, Lepocreadium setiferoides, Zoogonus rubellus, Austrobilharzia variglandis, and Gynaecotyla adunca infections were observed in recovered snails. At site A, in 1993, 62 natives were recovered. Among the 26 initially testing uninfected, 15 had infections at recovery. Of 36 that released cercariae, 26 had the infection initially indicated. Of sentinels released at site B in 1996, 3 of 16 had infections when recollected. One probably was infected before transplant; at least 2 were infected at site B during 1996-1997. Among site B natives, 26 were later examined by dissection in 1996; 22 had the infection status initially revealed. Some site B natives (n = 35) escaped recapture in 1996 but were found in 1997, or 1998, or both, and were reexamined, most by cercarial release. The same cercariae were produced by 30. Among natives (both sites, n = 123), 27.6% exhibited some difference in infection status compared to the initial assay. This probably overestimates changes. Some differences were real but most can be discounted as cases where initial infection status was misrepresented.     

Polyadenylation of nascent RNA during the embryogenesis of Ilyanassa obsoleta.

It has been demonstrated that specific changes in carbohydrate-containing cell surface lectin receptor sites occur with differentiation and maturation of sea urchin embryo cells. In this study, evidence is presented, using a quantitative electronic particle counter assay to measure agglutination, which indicates that concanavalin A (Con A) mediated agglutination of dissociated $\text{32}\text{64}$ cell sea urchin embryos differs dramatically with respect to specific cell populations. The migratory cell type, the micromere, is significantly more agglutinable with Con A than the other cell types and colchicine treatment markedly increases sea urchin embryo cell agglutinability. The results indicate that like many malignant cells which display extensive migratory behavior, specific migratory populations of embryonic cells are agglutinable with Con A. The results are discussed with respect to the possible nature of lectin receptor sites on specific populations of embryonic cells and the possible role of colchicine-sensitive structures in controlling the display patterns of these sites.     

Selective grazing by the mud snail Ilyanassa obsoleta

Summary Mud snails (Ilyanassa obsoleta) starved for 48 h were allowed to feed on sediments in laboratory microcosms. Sediment cores sliced at 2 mm intervals were compared to snail stomach contents for per cent carbon and nitrogen, plant pigment contents and species composition of benthic diatoms. Concentrations of carbon, nitrogen, phaeopigments, phycocyanin and chlorophyll were enriched in the top 2 mm of the sediments compared to 7–10 mm depth by a factor of 2–10. In turn, these materials were 20–40 times more concentrated in snail guts than in the surface sediments. Snail feces were enriched for carbon and nitrogen by 5–7 times over the surface sediments. Bacterial chlorophyll peaked at about 3–4 mm in the sediments and was not detectable in the snail stomach contents. The C/N ratio of the snail stomach contents was only 6 compared to a ratio of 8.5 for their feces and 12 for the surface sediments.The percentage of migratory diatoms (e.g. Nitzschia and Navicula) decreased with depth where non-migratory species, such as Fragilaria pinnata, dominated. These migratory species were more common in the snails than in the sediments on which they were feeding.A comparison of daily ingestion rates to the animal's energy budget shows that this selective ingestion is sufficient to meet Ilyanassa's energy needs.     

An unusual cytoplasmic organelle in oocytes of Ilyanassa obsoleta

An unusual organelle in the oocytes of the marine mud snail Ilyanassa obsoleta is described and called a polymerosome, because of its several components. Polymerosomes appear at the beginning of vitellogenesis and persist, in part, through postvitellogenesis. The persistent part is similar to an organelle that has been described by other workers in the vegetal region and the polar lobe of the Ilyanassa egg.     

Analysis of ciliary band formation in the mollusc Ilyanassa obsoleta

Two primary ciliary bands, the prototroch and metatroch, are required for locomotion and in the feeding larvae of many spiralians. The metatroch has been reported to have different cellular origins in the molluscs Crepidula fornicata and Ilyanassa obsoleta, as well as in the annelid Polygordius lacteus, consistent with multiple independent origins of the spiralian metatroch. Here, we describe in further detail the cell lineage of the ciliary bands in the gastropod mollusc I. obsoleta using intracellular lineage tracing and the expression of an acetylated tubulin antigen that serves as a marker for ciliated cells. We find that the I. obsoleta metatroch is formed primarily by third quartet derivatives as well as a small number of second quartet derivatives. These results differ from the described metatrochal lineage in the mollusc C. fornicata that derives solely from the second quartet or the metatrochal lineage in the annelid P. lacteus that derives solely from the third quartet. The present study adds to a growing body of literature concerning the evolution of the metatroch and the plasticity of cell fates in homologous micromeres in spiralian embryos.     

Regulation of Protein Synthesis in Embryos of Ilyanassa obsoleta

Much of the protein synthesis during early development in Ilyanassaresults from the translation of oogenetic mRNA. We show thatmicrotubule proteins are products of this translation, and thattheir synthesis is subject to translation level regulation.We also show that translation level regulation is involved inthe function of the polar lobe by making comparisons of theelectrophoretic patterns of synthesis of ¹⁴C labeled proteinsof normal embryos with the patterns of synthesis of ³H labeledproteins of embryos from which the polar lobes had been removedat the trefoil stage. Controls utilizing biochemical and morphologicalmarkers were performed to assure that normal and delobed embryoswere developing at equivalent rates. The expression of significantdifferences in the patterns of protein synthesis were foundbetween normal and delobed embryos, and these differences werenot dependent upon concomitant RNA synthesis. These differenceswere observable as early as after only 24 hours of development,although organogenesis does not begin until much later in development.Therefore, the observed differences probably reflect determinativeevents. The results support the hypothesis that the developmentaldeterminants of the polar lobe may include specific, preformedmRNA sequences, or specific regulators of translation.     

Fate maps of the first quartet micromeres in the gastropod Ilyanassa obsoleta.

Cell fate specification in the gastropod mollusc Ilyanassa obsoleta involves both cell autonomous and inductive mechanisms, which depend on determinants localized first in the polar lobe and then in the D quadrant of the embryo. A complete cell lineage is lacking for this embryo and is essential for a critical interpretation of previous experimental results and an analysis of the mechanisms at the molecular level. Lineages of the first quartet micromeres were followed using Lucifer Yellow dextran as a tracer. The tracer was injected into individual first quartet micromeres using iontophoresis and patterns of fluorescence were analyzed in the larva after 8 days of development. Fluorescence was limited to head structures, including eyes, tentacles and velum. Structures on the left side were derived from 1a and 1d micromeres; 1a gave rise to the left eye, including the lens. Right side structures were derived from the 1c micromere and 1b contributed to the apical plate between the eyes and symmetrically to both sides of the velum. First quartet lineage data are compared with results from previous cell ablation experiments and with lineage data from other species.     

Infrapopulation sizes of co-occurring trematodes in the snail Ilyanassa obsoleta

This study addresses the infrapopulation sizes of 2 larval trematode species Himasthla quissetensis and Zoogonus rubellus as they co-occur within their estuarine snail host Ilyanassa obsoleta. Rediae of H. quissetensis and sporocysts of Z rubellus were counted in snails singly infected with each parasite and in snails infected with both. Comparisons of the counts indicate that infrapopulations of H. quissetensis were unaffected by co-occurrence with Z rubellus. However, Z. rubellus infrapopulations were reduced when co-occurring with H. quissetensis. It is proposed that this situation does not result from an interspecific interaction between parasite species. Although this double infection is relatively frequent in certain snail populations, it is contended that these trematode species do not co-occur often enough to evolve responses to one another. However, the host environment must be encountered in each life cycle, and both trematode species must be adapted to use it. On this basis, whatever happens when these 2 species occupy the same host is based on adaptations of the parasites to the host. It is proposed that these parasites are adapted to self-limit their infrapopulations in the snail host. They can, thus, preserve and use the host for many years and thereby enhance total cercarial transmission (fitness). Infrapopulation sizes would be determined by host resource levels, which, among other factors, would be influenced by the presence of multiple parasite species. In single infections, by far the most common situation, host resource levels would be set by the nutritional status or age (size) of the host (or both). The reduced infrapopulation sizes of Z rubellus on co-occurrence suggest that this trematode is more sensitive to host resource levels than is H. quissetensis.     

Polar lobe specific regulation of translation in embryos of Ilyanassa obsoleta.

Comparisons of the patterns of synthesis of ¹⁴C-labeled proteins of normal embryos of Ilyanassa with the patterns of synthesis of ³H-labeled proteins of embryos from which the polar lobes had been removed at the trefoil stage were made by coelectrophoresis on polyacrylamide disc gels. Controls utilizing biochemical and morphological markers were performed to assure that normal and delobed embryos were developing at equivalent rates. The expression of significant differences in the patterns of protein synthesis were found between normal and delobed embryos, and these differences were not dependent upon concomitant RNA synthesis. These differences were observable as early as after only 24 hr of development, although organogenesis does not begin until much later in development. Therefore, the observed differences probably reflect determininative events. The results support the hypothesis that the developmental determinants of the polar lobe may include specific, preformed mRNA sequences, or specific regulators of translation.     

Serotonin and Nitric Oxide Regulate Metamorphosis in the Marine Snail Ilyanassa obsoleta

Several neuroactive compounds have been implicated as playingroles in the circuitry that controls larval metamorphosis inmarine molluscs. For the caenogastropod Ilyanassa obsoleta,results of neuroanatomical studies suggest that the productionof nitric oxide (NO) increases throughout the planktonic stageand that NO production is necessary for the maintenance of thelarval state, especially as it becomes metamorphically competent.Bath application or injection of exogenous serotonin (5HT) caninitiate metamorphosis in competent larvae, and exogenous NOcan inhibit such serotonergically-induced metamorphosis. Inhibitionof endogenous nitric oxide synthase (NOS) can also trigger larvalmetamorphosis. The production of endogenous NO appears to decreaseconcurrently with the initiation of metamorphosis, but the specificinteractions between serotonergic and nitrergic neurons areunknown. Evidence in support of NO acting to up-regulate theenzyme guanylyl cyclase (GC) is still equivocal. Thus, we donot yet know if NO exerts its effects through the actions ofcyclic 3',5'-guanosine monophosphate (cGMP) or by a cGMP-independentmechanism. The ubiquity of nitrergic signalling and its significancefor developing molluscan embryos and larvae are still the subjectof speculation and require further investigation.     

Nucleic Acid Chemistry of the Ilyanassa Embryo

A morphological study of Ilyanassa embryogenesis was made fromsections of several stages and the nucleic acid content wasmeasured throughout the course of development. The kineticsof uptake and incorporation of radioactive precursors into RNAwas determined. The RNAs synthesized at different stages ofembryogenesis were fractionated by methylated albumin kieselguhrchromatography, and the polyadenylation of RNA during developmentwas measured. From these studies it was concluded that the Ilyanassa embryo,beginning with the 4-cell stage, synthesizes dRNA, rRNA, 4SRNA, and 5S RNA throughout all stages of development. The extensivesynthesis of dRNA and the higher proportion of RNAs polyadenylatedby the pregastrula embryo was considered to be of particularsignificance. The localization of nucleic acids and nucleic acid precursorsin the polar lobe and the effect of removing the polar lobeon nucleic synthesis was reviewed. Recent work on the ultrastructure of the Ilyanassa egg was reviewedin relation to the problem of ooplasmic segregation. The discoveryof a new cytoplasmic organelle in the Ilyanassa oocyte was reviewed.