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TheGulfofGabesintheEasternMediterraneanBasinisoneofthelargestcontinentalshelvesandisahighly productivearea.ThisisincontrasttotheEasternMediterraneanBasinasawholewhichisoligotrophicto ultraoligotrophic.Overrecentyears,duetoincreasedpressurefromanthropologicalactivities,therehas beenadeclineinthebiologicalresourcesintheGulfofGabes.Thishasledscientiststofocustheirresearch on thedifferenttrophiclevelsofthefoodchainandinparticularonphytoplankton.Studiesdedicatedto phytoplanktonintheGulfofGabeshavepreviouslyreliedonconventionalglobalapproaches.Thisworkon theotherhand,examinesultraphytoplankton(o10 mm) distributionatthesinglecelllevelindifferent watermasses,duringasummercruise(2008).Threedistinctwatermasseswereidentified:modified Mediterraneanwaters(MMW),modified Atlanticwaters(MAW)andIonianwaters(IW).Despitethe summerperiod,nutrientswereabundantandnon-limiting.Flowcytometryresolved five cellgroups, Prochlorococcus, Synechococcus, picoeukaryotes,nanoeukaryotesandagroupofunidentified cells.These cellswerecharacterisedbylow fluorescencesignalsandscattersignalslargerthanthatofcyanobacteriaand picoeukaryotesandthuswerelabelledLFNanoforlow fluorescencenanoeukaryotes.Theywerefoundin veryhighabundances,whereastheotherusualgroupswererelativelylowinabundancedespitethe favourablegrowthconditions.LFNanofoundduringa2012summercruiseinthesamearea,werealso sorted by flowcytometryandobservedbyscanningelectronmicroscopy.Choanoflagellateswereidentified, andthesenanoflagellatesappeartoaccountforthelowabundanceintheusualultraphytoplanktongroup due totheirefficientfeeding/grazingmechanism.However,themostabundantLFNanocouldnotbe identified.Resultsstronglysuggestatop-downcontroloftheecosystem,consistentwiththehigh productivityoftheGulfthatimpliesarapidtransferofmatterandenergytohighertrophiclevels.

Embryonic development from coeloblastula to fully developed larva was investigated in 8 Mediterranean homoscleromorph species: Oscarella lobularis, O. tuberculata, O. microlobata, O. imperialis, Plakina trilopha, P. jani, Corticium candelabrum, and Pseudocorticium jarrei. Morphogenesis of the larva is similar in all these species; however, cell proliferation is more active in species of Oscarella than in Plakina and C. candelabrum. The result of cell division is a wrinkled, flagellated larva, called a cinctoblastula. It is composed of a columnar epithelium of polarized, monoflagellated cells among which are scattered a few non-flagellated ovoid cells. The central cavity always contains symbiotic bacteria. Maternal cells are also present in O. lobularis, O. imperialis, and P. jarrei. In the fully developed larva, cell shape and dimensions are constant for each species. The cells of the anterior pole have large vacuoles with heterogeneous material; those of the postero-lateral zone have an intranuclear paracrystalline inclusion; and the flagellated cells of the posterior pole have large osmiophilic inclusions. Intercellular junctions join the apical parts of the cells, beneath which are other specialized cell junctions. A basement membrane underlying the flagellated cells lines the larval cavity. This is the first observation of a basement membrane in a poriferan larva. The basal apparatus of flagellated cells is characterized by an accessory centriole located exactly beneath the basal body. The single basal rootlet is cross striated. The presence of a basement membrane and a true epithelium in the larva of Homoscleromorpha—unique among poriferan clades and shared with Eumetazoa—suggests that Demospongiae could be paraphyletic.

Summary Oogenesis, spermatogenesis, embryogenesis, and the structure of the larva of Oopsacas minuta (Hexactinellida, Hexasterophora) were studied in a population living in a Mediterranean cave and which reproduces year round. The different stages in oogenesis and the total and equal cleavage of the zygote are similar to the same events in the Calcarea and Demospongiae. During early development, the embryo is clearly cellular. The first stages of cleavage are spiral, and gastrulation occurs by primary delamination. These features are unique in the phylum Porifera where the occurrence of true gastrulation is controversial. The larva possesses multi-flagellated cells that are linked by parallel junctions, as well as larval spicules, choanochambers, and symbiotic bacteria. Part of larval tissue is syncytial. Spiculogenesis is intrasyncytial rather than extracellular as in the Calcarea, and the ultrastructure of the sclerocyte is similar to that of Demospongiae except that it is multinucleate. The development...

Only three species of the sponge genus Plakina Schulze have been described from the Mediterranean since 1880, in spite of a large amount of allegedly intraspecific variability in morphological characters. However, recent genetic studies based on electrophoretic techniques have revealed extensive cryptic speciation in north-western Mediterranean Plakina, demonstrating that most of this variation was interspecific rather than intraspecific. We describe in detail the morphology and anatomy of four new species of Plakina from the Mediterranean–P. crypta, P. weinbergi, P. endoumensis and P. jani–of which the latter two were discovered through allozyme electrophoresis. Plakina monolopha Schulze and P. trilopha Schulze are redescribed, and their morphological and geographical limits are discussed along with those of P. dilopha Schulze. Accurate analysis of the internal anatomy and of the shape and ramification pattern of lophose spicules in scanning electron microscopy provides new, powerful morphological criteria for species discrimination in Plakina. More widespread use of such new taxonomic characters should provide evidence against the alleged cosmopolitanism of some Plakina species, thus generating an increase in estimates of the biodiversity of plakinids.

Three new sponge species without a skeleton, Oscarella viridis, O. microlobata, and O. imperialis, were found in sublittoral caves and on vertical walls along the coast of Provence (western Mediterranean Sea, France). Their morphology, anatomy, and cytology are described and they are compared with the two other valid Mediterranean Oscarella species, O. lobularis and O. tuberculata. Reproductive and internal anatomical characters are uniform in the genus, but details of external morphology and especially cytological characters (mesohylar cells with inclusions) provide good diagnostic features at the species level. Careful observation of morphological and cytological characters is essential for clarifying the systematics of Oscarella and reveals an unexpected biodiversity of this genus in the Mediterranean Sea.

International audience; Sponges branch basally in the metazoan phylogenetic tree and are thus well positioned to provide insights into the evolution of mechanisms controlling animal development, likely to remain active in adult sponges. Of the four sponge clades, the Homoscleromorpha are of particular interest as they alone show the ''true'' epithelial organization seen in other metazoan phyla (the Eumetazoa). We have examined the deployment in sponges of Wnt signalling pathway components, since this pathway is an important regulator of many developmental patterning processes. We identified a reduced repertoire of three divergent Wnt ligand genes in the recently-sequenced Amphimedon queenslandica (demosponge) genome and two Wnts from our EST collection from the homoscleromorph Oscarella lobularis, along with well-conserved genes for intracellular pathway components (b-catenin, GSK3b). Remarkably, the two O. lobularis Wnt genes showed complementary expression patterns in relation to the evenly spaced ostia (canal openings) of the exopinacoderm (ectoderm), highly reminiscent of Wnt expression during skin appendage formation in vertebrates. Furthermore, experimental activation of the Wnt/b-catenin pathway using GSK3b inhibitors provoked formation of ectopic ostia, as has been shown for epithelial appendages in Eumetazoa. We thus suggest that deployment of Wnt signalling is a common and perhaps ancient feature of metazoan epithelial patterning and morphogenesis. Citation: Lapébie P, Gazave E, Ereskovsky A, Derelle R, Bézac C, et al. (2009) WNT/b-Catenin Signalling and Epithelial Patterning in the Homoscleromorph Sponge Oscarella. PLoS ONE 4(6): e5823.

The metamorphosis of the cinctoblastula of Homoscleromorpha is studied in five species belonging to three genera. The different steps of metamorphosis are similar in all species. The metamorphosis occurs by the invagination and involution of either the anterior epithelium or the posterior epithelium of the larva. During metamorphosis, morphogenetic polymorphism was observed, which has an individual character and does not depend on either external or species specific factors. In the rhagon, the development of the aquiferous system occurs only by epithelial morphogenesis and subsequent differentiation of cells. Mesohylar cells derive from flagellated cells after ingression. The formation of pinacoderm and choanoderm occurs by the differentiation of the larval flagellated epithelium. This is possibly due to the conservation of cell junctions in the external surface of the larval flagellated cells and of the basement membrane in their internal surface. The main difference in homoscleromorph metamorphosis compared with Demospongiae is the persistence of the flagellated epithelium throughout this process and even in the adult since exo- and endopinacoderm remain flagellated. The antero-posterior axis of the larva corresponds to the baso-apical axis of the adult in Homoscleromorpha.

Data on nonbilaterian animals (sponges, cnidarians, and ctenophores) have suggested that Antennapedia (ANTP) class homeobox genes played a crucial role in the early diversification of animal body plans. Estimates of ancestral gene diversity within this important class of developmental regulators have been mostly based on recent analyses of the complete genome of a demosponge species, leading to the proposal that all ANTP families found in nonsponges animals (eumetazoans) derived from an ancestral “proto-NK” six-gene cluster. However, a single sponge species cannot reveal ancestral metazoan traits, in particular because lineage-specific gene duplications or losses are likely to have occurred during the long history of the Porifera. We thus looked for ANTP genes by degenerate polymerase chain reaction search in five species belonging to the Homoscleromorpha, a sponge lineage recently phylogenetically classified outside demosponges and characterized by unique histological features. We identified new genes of the ANTP class called HomoNK. Our phylogenetic analyses placed HomoNK (without significant support) close to the NK6 and NK7 families of cnidarian and bilaterian ANTP genes and did not recover the monophyly of the proposed “proto-NK” cluster. Our expression analyses of the HomoNK gene OlobNK in adult Oscarella lobularis showed that this gene is a strict marker of choanocytes, the most typical sponge cell type characterized by an apical flagellum surrounded by a collar of microvilli. These results are discussed in the light of the predominant neurosensory expression of NK6 and NK7 genes in bilaterians and of the recent proposal that choanocytes could be the sponge homologs of sensory cells.