Sexual reproduction of the placental brooder Celleporella hyalina (Bryozoa, Cheilostomata) in the White Sea.

The evolution of parental care is a central field in many ecological and evolutionary studies, but integral approaches encompassing various life-history traits are not common. Else, the structure, development and functioning of the placental analogues in invertebrates are poorly understood. Here, we describe the life-history, sexual colony dynamics, oogenesis, fertilization and brooding in the boreal-Arctic cheilostome bryozoan Celleporella hyalina. This placental brooder incubates its progeny in calcified protective chambers (ovicells) formed by polymorphic sexual zooids. We conducted a detailed ultrastructural study of the ovary and oogenesis, and provide evidence of both auto- and heterosynthetic mechanisms of vitellogenesis. We detected sperm inside the early oocyte and within funicular strands, and discuss possible variants of fertilization. We also detail the development and functioning of the placental analogue (embryophore) in the various stages of embryonic incubation as well as embryonic histotrophic nourishment. In contrast to all known cheilostome placentas, the main part of embryophore of C. hyalina is not a single cell layer. Rather, it is a massive "nutritive tissue" whose basal part is associated with funicular strands presumably providing transport function. C. hyalina shows a mixture of reproductive traits with macrolecithal oogenesis and well-developed placenta. These features give it an intermediate position in the continuum of variation of matrotrophic provisioning between lecithotrophic and placentotrophic cheilostome brooders. The structural and developmental differences revealed in the placental analogue of C. hyalina, together with its position on the bryozoan molecular tree, point to the independent origin of placentation in the family Hippothoidae.

Cleavage modification did not alter blastomere fates during bryozoan evolution.

BACKGROUND: Stereotypic cleavage patterns play a crucial role in cell fate determination by precisely positioning early embryonic blastomeres. Although misplaced cell divisions can alter blastomere fates and cause embryonic defects, cleavage patterns have been modified several times during animal evolution. However, it remains unclear how evolutionary changes in cleavage impact the specification of blastomere fates. Here, we analyze the transition from spiral cleavage - a stereotypic pattern remarkably conserved in many protostomes - to a biradial cleavage pattern, which occurred during the evolution of bryozoans. RESULTS: Using 3D-live imaging time-lapse microscopy (4D-microscopy), we characterize the cell lineage, MAPK signaling, and the expression of 16 developmental genes in the bryozoan Membranipora membranacea. We found that the molecular identity and the fates of early bryozoan blastomeres are similar to the putative homologous blastomeres in spiral-cleaving embryos. CONCLUSIONS: Our work suggests that bryozoans have retained traits of spiral development, such as the early embryonic fate map, despite the evolution of a novel cleavage geometry. These findings provide additional support that stereotypic cleavage patterns can be modified during evolution without major changes to the molecular identity and fate of embryonic blastomeres.

Phylogenetically Widespread Polyembryony in Cyclostome Bryozoans and the Protracted Asynchronous Release of Clonal Brood-Mates.

Polyembryony-the production of multiple cloned embryos from a single fertilised egg-is a seemingly paradoxical combination of reproductive modes that nevertheless persists in diverse taxa. We document features of polyembryony in the Cyclostomata (Bryozoa)-an ancient order of modular colonial marine invertebrates-that suggest a substantial reduction in the paradoxical nature of this enigmatic reproductive mode. Firstly, we provide molecular evidence for polyembryony in three exemplar species, supporting the widely cited inference that polyembryony characterises the entire order. Secondly, genotyping demonstrates protracted release of cloned offspring from the primary embryo in a given gonozooid (chamber for embryonic incubation), thus exposing the same genotype to changing environmental conditions over time. Finally, we confirm that each gonozooid produces a distinct genotype, with each primary embryo being the result of a separate fertilisation event. We hypothesise that the sustained release of one or a few genotypes against varying environmental conditions achieves levels of risk-spreading similar to those in organisms that release multiple, unique genotypes at a single time. We argue that polyembryony, specifically with the production of a large number of progeny per fertilisation event, has been favoured in the Cyclostomata over long geological periods.

From incipient to substantial: evolution of placentotrophy in a phylum of aquatic colonial invertebrates.

Matrotrophy has long been known in invertebrates, but it is still poorly understood and has never been reviewed. A striking example of matrotrophy (namely, placentotrophy) is provided by the Bryozoa, a medium-sized phylum of the aquatic colonial filter feeders. Here I report on an extensive anatomical study of placental analogues in 21 species of the bryozoan order Cheilostomata, offering the first review on matrotrophy among aquatic invertebrates. The first anatomical description of incipient placentotrophy in invertebrates is presented together with the evidence for multiple independent origins of placental analogues in this order. The combinations of contrasting oocytic types (macrolecithal or microlecithal) and various degrees of placental development and embryonic enlargement during incubation, found in different bryozoan species, are suggestive of a transitional series from the incipient to the substantial placentotrophy accompanied by an inverse change in oogenesis, a situation reminiscent of some vertebrates. It seems that matrotrophy could trigger the evolution of sexual zooidal polymorphism in some clades. The results of this study show that this phylum, with its wide variety of reproductive patterns, incubation devices, and types of the simple placenta-like systems, offers a promising model for studying parallel evolution of placentotrophy in particular, and matrotrophy in general.

Effects of selfing on offspring survival and reproduction in a colonial simultaneous hermaphrodite (Bugula stolonifera, Bryozoa).

Understanding the consequences of selfing in simultaneous hermaphrodites requires investigating potential deleterious effects on fitness at all stages of life. In this study, I examined the effects of selfing throughout the life cycle of the marine bryozoan Bugula stolonifera, a colonial simultaneous hermaphrodite. In 2008, larvae from field-collected colonies were cultured through metamorphosis to reproductively mature colonies either in the presence of one other colony, the paired treatment, or alone, the solitary treatment. Results demonstrated that selfing in this species is possible, in that colonies in the solitary treatment produced viable larvae that successfully completed metamorphosis. On average, however, these colonies released significantly fewer larvae, which experienced reduced rates of metamorphic initiation and completion compared to the paired treatment. These experiments were extended in 2009, when metamorphs from colonies reared in the solitary (n = 58) and paired (n = 61) treatments were transferred to the field for growth to reproductive maturity and then brought back to the laboratory for larval collection. Results revealed additional deleterious effects associated with selfing, as no viable larvae were recovered from colonies deriving from the solitary treatment. In contrast, offspring from the paired treatment released 1030 larvae and 99% initiated metamorphosis, 97% of which completed metamorphosis. Overall, selfed larvae not only had significantly decreased chances of survival, but those that did survive did not successfully reproduce.

Does the survivorship of activated resting stages in toxic environments provide cues for ballast water treatment?

The toxic effects of three inorganic metals (Cu, Cr, Hg), three organic (phenol, formalin, ammonium) chemicals, ozone-enriched water and peroxides (H2O2) on embryonic development were tested in 8 species from the Porifera, Bryozoa and Crustacea. Toxicants with lower molecular weight showed stronger negative impacts on post-diapause embryos than chemicals with higher molecular weight if related to the toxicity of the chemicals to active adult stages. Only few embryos of the cladoceran Moina macrocopa and none of the cladoceran Wlassicsia pannonica treated with peroxides at concentration 0.3% developed further. Ozone-enriched water had no significant effect on post-diapause embryonic development in cladocerans. Ammonium (the product of NH4OH dissociation) in concentration 100 mg/l and higher killed all embryos of M. macrocopa inside protective membranes. Peroxides and ammonium are suggested for the purification of ship ballast waters as effective, non-expensive and non-persistent toxic chemicals. Resting stages of invertebrates including at least Crustaceans, Porifera and Bryozoa seem to allow not only dispersal among toxic industrial environments such as ship ballast compartments, but may also endure serious pollution events common in seaports and estuaries. Artemia cysts due to their strong protection against different toxic substances are recommended as a model for studies of toxic effects in diapausing stages in polluted estuaries and marine environments.

The internal-brooding apparatus in the bryozoan genus Cauloramphus (Cheilostomata: Calloporidae) and its inferred homology to ovicells.

We studied by SEM the external morphology of the ooecium in eight bryozoans of the genus Cauloramphus (Cheilostomata, Calloporidae): C. spinifer, C. variegatus, C. magnus, C. multiavicularia, C. tortilis, C. cryptoarmatus, C. niger, and C. multispinosus, and by sectioning and light microscopy the anatomy of the brooding apparatus of C. spinifer, C. cryptoarmatus, and C. niger. These species all have a brood sac, formed by invagination of the non-calcified distal body wall of the maternal zooid, located in the distal half of the maternal (egg-producing) autozooid, and a vestigial, maternally budded kenozooidal ooecium. The brood sac comprises a main chamber and a long passage (neck) opening externally independently of the introvert. The non-calcified portion of the maternal distal wall between the neck and tip of the zooidal operculum is involved in closing and opening the brood sac, and contains both musculature and a reduced sclerite that suggest homology with the ooecial vesicle of a hyperstomial ovicell. We interpret the brooding apparatus in Cauloramphus as a highly modified form of cheilostome hyperstomial ovicell, as both types share 1) a brood chamber bounded by 2) the ooecium and 3) a component of the distal wall of the maternal zooid. We discuss Cauloramphus as a hypothetical penultimate stage in ovicell reduction in calloporid bryozoans. We suggest that the internal-brooding genus Gontarella, of uncertain taxonomic affinities, is actually a calloporid and represents the ultimate stage in which no trace of the ooecium remains. Internal brooding apparently evolved several times independently within the Calloporidae.

Comparative anatomical study of internal brooding in three anascan bryozoans (Cheilostomata) and its taxonomic and evolutionary implications.

The anatomical structure of internal sacs for embryonic incubation was studied using SEM and light microscopy in three cheilostome bryozoans-Nematoflustra flagellata (Waters,1904), Gontarella sp., and Biflustra perfragilis MacGillivray, 1881. In all these species the brood sac is located in the distal half of the maternal (egg-producing) autozooid, being a conspicuous invagination of the body wall. It consists of the main chamber and a passage (neck) to the outside that opens independently of the introvert. There are several groups of muscles attached to the thin walls of the brood sac and possibly expanding it during oviposition and larval release. Polypide recycling begins after oviposition in Gontarella sp., and the new polypide bud is formed by the beginning of incubation. Similarly, polypides in brooding zooids degenerate in N. flagellata and, sometimes, in B. perfragilis. In the evolution of brood chambers in the Cheilostomata, such internal sacs for embryonic incubation are considered a final step, being the result of immersion of the brooding cavity into the maternal zooid and reduction of the protecting fold (ooecium). Possible reasons for this transformation are discussed, and the hypothesis of Santagata and Banta (Santagata and Banta1996) that internal brooding evolved prior to incubation in ovicells is rejected.

Paradoxical polyembryony? Embryonic cloning in an ancient order of marine bryozoans.

Prolific polyembryony is reported in few major taxa, but its occurrence has generated theoretical debate on potential conflict between sexual and asexual reproduction. It is, therefore, important to genetically confirm a widely cited inference, based on microscopy, that polyembryony characterizes marine bryozoans of the order Cyclostomata. Microsatellite genotyping of brooded embryos and maternal colonies conclusively demonstrated polyembryony, while genetic variation among broods within colonies indicated outcrossing via water-borne sperm, in the rocky-shore species Crisia denticulata. The characteristically voluminous brood chamber of cyclostomes is judged to be an adaptation linked to larval cloning and hence an indicator of polyembryony. We speculate that although the almost universal occurrence of polyembryony among crown-group Cyclostomata is probably attributable to phylogenetic constraint, adaptive consequences are likely to be significant.

[Systematic value of the larval structure and details of postlarval morphogenesis in Bryozoa gymnolaemates]. / Valeur systématique de la structure larvaire et des particularités de la morphogenèse post-larvaire chez les Bryozoaires Gymnolaemates.

Among the various species of Bryozoa Gymnolaemata, the larvae and their development were studied, comparing the larval structure and the evolution of their cellular categories during the post-larval morphogenesis the existence of nine well-defined larval types could be revealed. Cases of insufficiently described larvae are discussed. The present systematic of Bryozoa Gymnolaemata is compared with the classification of various larval types. For the major part of cases, each systematic family is marked by a precise type of larva; however there are some exceptions, especially in the ordre Ctenostomata. These discordances may suggest some rearrangements of the classification utilized at the present time.