Initial stages of food ingestion by Sepia officinalis (Mollusca: Cephalopoda)

The cuttlefish ingests much skeleton from the crustaceans and fish it preys upon. The skeletal pieces are relatively large and their dimensions bear a close relationship to the length of the buccal mass and diameter of the oesophagus. The structures of the buccal mass are instrumental in the breakdown of prey and orientation of long pieces of skeleton to ensure their entry into the oesophagus. Many pieces of skeletal material present in the stomach contents still have attached muscles, showing that there is little, or no, external digestion. Skeletal material may be important for long-term maintenance of young Sepia in captivity.     

Serotonergic regulation of the central heart auricles of Sepia officinalis L. (Mollusca, Cephalopoda)

In pharmacological bioassays on isolated isotonically suspended auricles of Sepia officinalis, the regulatory action of the neurotransmitter serotonin (5-hydroxytryptamine, 5-HT) on these autonomous contractile compartments was demonstrated. 5-HT induced concentration-dependent positive effects on frequency and tone, whereas the concentrations/response curve for the amplitude showed a biphasic course. All applied antagonists inhibited mainly the effect of 5-HT on frequency and amplitude. The chronotropic effects of 5-HT were blocked mainly by the 5-HT(1,2) antagonist methiothepin (pA(2)=8.01), the 5-HT(1a) antagonist NAN-190 (pA(2)-) and in lesser extent by the 5-HT(1,2) antagonist mianserin (pA(2)=6.81). In the presence of each antagonist applied the 5-HT action on amplitude was transformed into a positive inotropic effect with the maximum under influence of NAN-190 and the 5-HT(2,1c)-antagonist ketanserin. The auricular tone was also influenced by the antagonists and in combination with methiothepin it turned into strong negative tonotropic effect. In addition to the pharmacological bioassays, the presence of 5-HT in nerve endings within the auricle wall was demonstrated by immunohistochemical and fluorescence microscopic findings. Altogether the findings presented here confirm that 5-HT evokes excitatory effects on the autonomous contractile auricle of S. officinalis and acts obviously over different receptors, whereby a 5-HT(1)- and a 5-HT(2)-like seem to be involved.     

The cytoskeleton of chondrocytes of Sepia officinalis (Mollusca, Cephalopoda): an immunocytochemical study

Our previous electron microscope study showed that chondrocytes from cephalopod cartilage possess a highly developed cytoskeleton and numerous cytoplasmic processes that ramify extensively through the tissue. We have now carried out a light microscope immunocytochemical study of chondrocytes from the orbital cartilage of Sepia officinalis to obtain indications as to the nature of the cytoskeletal components. We found clear positivity to antibodies against mammalian tubulin, vimentin, GFAP, and actin, but not keratin. The simultaneous presence of several cytoskeletal components is consistent with the hypothesis that cephalopod chondrocytes have the characteristics of both chondrocytes and osteocytes of vertebrates, which endow the tissue as a whole with some of the properties of vertebrate bone. We confirm, therefore, the presence in molluscs of the ubiquitous cytoskeletal proteins of metazoan cells that have remained highly conserved throughout phylogenetic evolution.     

The ultrastructure of chondrocytes in the cartilage of Sepia officinalis and Octopus vulgaris (Mollusca, Cephalopoda)

Thin sections of cartilage from the chondrocranium of cuttle fish and octopus were examined using the transmission electron microscope. It was found that cephalopod chondrocytes differed considerably from the chondrocytes of vertebrate cartilage; in particular they possessed many long and ramifying cytoplasmic processes and had an ultrastructure typical of protein-secreting cells. They did not, however, contain secretory granules; while vesicles and rough endoplasmic reticulum cisternae seemed to open directly to the cell surface. The cell body and processes contained cytoskeletal structures: microtubules were easily recognized, but intermediate and thin filaments were difficult to make out as they were frequently clumped into bundles. Some chondrocytes contained conspicuous accumulations of hemocyanin. The cytoplasmic processes possessed intercellular contacts similar to gap junctions. Also present on processes and the cell body were cell-extracellular matrix focal adhesions. The chondrocytes were not polarized or arranged in any preferential spatial order, however, with their processes they formed a three-dimensional network throughout the cartilage tissue. Ultrastructural findings are discussed in relation to the singular morphofunctional characteristics of cephalopod cartilage which shares features with both the cartilage and bone of vertebrates.     

Fine structure and function of the digestive cell of Sepia officinalis (Mollusca: Cephalopoda)

The post–embryonic development of the digestive gland has been studied by light and electron microscopy on animals reared in the laboratory. The gland reaches its "adult" structure around the end of the first month after hatching.
The infrastructural changes of the digestive gland have been followed during digestion at various ages (5,10,20,30 days and "adult"); ferritin was used for a tracer to study absorption.
These experiments have produced evidence of endocytosis intake of large proteins and further intracellular digestion inside the digestive cell. The residues of digestion are excreted in a "brown body" while the metabolites are used in new syntheses for the cell itself. These results demonstrate that "ancestral" digestive processes have been kept in at least one cephalopod.     

orthodenticle/otx ortholog expression in the anterior brain and eyes of Sepia officinalis (Mollusca, Cephalopoda)

The origin of cerebral structures is a major issue in both developmental and evolutionary biology. Among Lophotrochozoans, cephalopods present both a derived nervous system and an original body plan, therefore they constitute a key model to study the evolution of nervous system and molecular processes that control the neural organization. We characterized a partial sequence of an ortholog of otx2 in Sepia officinalis embryos, a gene specific to the anterior nervous system and eye development. By in situ hybridization, we assessed the expression pattern of otx2 during S. officinalis organogenesis and we showed that otx is expressed (1) in the eyes, from early to late developmental stages as observed in other species (2) in the nervous system during late developmental stages. The otx ortholog does not appear to be required for the precocious emergence of the nervous ganglia in cephalopods and is later expressed only in the most anterior ganglia of the future brain. Finally, otx expression becomes restricted to localized part of the brain, where it could be involved in the functional specification of the central nervous system of S. officinalis. These results suggest a conserved involvement of otx in eye maturation and development of the anterior neural structures in S. officinalis.     

Components of the cellular defense and detoxification system of the common cuttlefish Sepia officinalis (Mollusca,Cephalopoda)

Endocytotic-active cells in the branchial heart complex of Sepia officinalis were studied by in situ injection of different types of xenobiotics and by in vitro perfusion of the organ complex with a bacterial suspension. The rhogocytes (ovoid cells) ingest particles of all tested sizes by endocytosis and phagocytosis. The hemocytes of the circulating blood and the adhesive hemocytes in the wall of the branchial heart incorporate all tested kinds of foreign materials, including bacterial cells due to phagocytosis achieved by the triangular mesenchymatic cells. The ultrastructural findings also give strong evidence that the triangular mesenchymatic cells are fixed hemocytes that have migrated into the branchial heart tissue. The ingestion and digestion of allogeneic substances and bacteria or their debris by rhogocytes and/or all (forms of) hemocytes suggests the involvement of these either fixed or mobile endocytotic-active cells in the defense and detoxification system of cephalopods.     

The stalked eyes of Bathothauma (Mollusca, Cephalopoda)

In the larvae of some cranchiid squids the eyes are carried sideways on long stalks and also possess a presumed luminous organ. The structure of this organ is described but the only sign of light producing tissues is some possible luminescent bacteria. The stalked eye may be connected with plankton feeding at great depths, perhaps improving distance judgement. The older animals of these species carry a different form of luminous organ.     

Food of the cuttlefish Sepia officinalis and S. elegans in the Ria de Vigo (NW Spain) (Mollusca: Cephalopoda)

A study of the diet of Sepia officinalis and S. elegans in the Ria de Vigo has shown that crustaceans are the most abundant prey in both species, followed by fish. Changes in the food composition of both species occur with growth. The type of prey eaten by the two sexes of these species is very similar. The possibility of trophic competition between juveniles of S. qficinalis and S. elegans is discussed.     

A new species of Dicyemennea Whitman, 1883 (Phylum Dicyemida) from Sepia latimanus (Mollusca: Cephalopoda: Decapodidae) off Okinawa, Japan

A new species of dicyemid mesozoan is described from a cuttlefish Sepia latimanus Quoy and Gaimard collected off Nago, Naha and Onnason, Okinawa Islands, Ryukyu Islands, Japan. Dicyemennea ryukyuense n. sp. is a large species that reaches about 5 mm in length. The vermiform stage is characterised by 23 peripheral cells, a conical calotte and an axial cell that extends to the base of the propolar cells. The infusoriform embryo consists of 37 cells; two nuclei are present in each urn cell and the refringent bodies are solid. In the type of reproductive strategy, rhombogens of D. ryukyuense form a small number of infusorigens and produce a relatively large number of gametes per infusorigen. In the family Dicyemidae, Dicyemennea Whitman, 1883 is the largest group after Dicyema von Kölliker, 1849. Other dicyemid species, including those belonging to Dicyema, were not detected. Dicyemids have never previously been detected in cephalopods living on corals and rocks off the Ryukyu Islands, even though they are benthonic in habitat. This is the first report of a dicyemid mesozoan from S. latimanus and also from off the Ryukyu Islands. S. latimanus inhabits coral reefs, often swimming over the coral but usually lying on the sandy bottom. Dicyemids cannot possibly infect cephalopods which live exclusively over coral and rock.     

Functional morphology of the mantle of Nautilus pompilius (Mollusca, Cephalopoda)

This study presents histological and cytological findings on the structural differentiation of the mantle of Nautilus pompilius in order to characterize the cells that are responsible for shell formation. The lateral and front mantle edges split distally into three folds: an outer, middle, and inner fold. Within the upper part of the mantle the mantle edge is divided into two folds only; the inner fold disappears where the hood is attached to the mantle. At the base of the outer fold of the lateral and front mantle edge an endo-epithelial gland, the mantle edge gland, is localized. The gland cells are distinguished by a distinct rough endoplasmic reticulum and by numerous secretory vesicles. Furthermore, they show a strong accumulation of calcium compounds, indicating that the formation of the shell takes place in this region of the mantle. Numerous synaptic contacts between the gland cells and the axons of the nerve fibers reveal that the secretion in the area of the mantle edge gland is under nervous control. The whole mantle tissue is covered with a columnar epithelium having a microvillar border. The analyses of the outer epithelium show ultrastructural characteristics of a transport active epithelium, indicating that this region of the mantle is involved in the sclerotization of the shell. Ultrastructural findings concerning the epithelium between the outer and middle fold suggest that the periostracum is formed in this area of the mantle, as it is in other conchiferan molluscs.     

Extensive population subdivision of the cuttlefish Sepia officinalis (Mollusca: Cephalopoda) around the Iberian Peninsula indicated by microsatellite DNA variation

The Atlantic Ocean-Mediterranean Sea junction has been proposed as an important phylogeographical area on the basis of concordance in genetic patterns observed at allozyme, mtDNA and microsatellite DNA markers in several marine species. This study presents microsatellite DNA data for a mobile invertebrate species in this area, the cuttlefish Sepia officinalis, allowing comparison of this relatively new class of DNA marker with previous allozyme results, and examination of the relative effects on gene flow of the Strait of Gibraltar and the Almería-Oran oceanographic front. Genetic variation at seven microsatellite loci screened in six samples from NE Atlantic and Mediterranean coasts of the Iberian Peninsula was high (mean Na = 9.6, mean H(e) = 0.725). Microsatellites detected highly significant subpopulation structuring (F(ST)= 0.061; R(ST) = 0.104), consistent with an isolation-by-distance model of low levels of gene flow. Distinct and significant clinal changes in allele frequencies between Atlantic and Mediterranean samples found at five out of seven loci, however indicate these results might be also consistent with an alternative model of secondary contact and introgression between previously isolated and divergent populations, as previously proposed for other marine species from the Atlantic-Mediterranean area. A pronounced 'step' change between SW Mediterranean samples associated with the Almería-Oran front suggests this oceanographic feature may represent a contemporary barrier to gene flow.     

Phylogeny of the Cocculinoidea (Mollusca, Gastropoda)

Abstract. The superfamily Cocculinoidea is a group of marine, deep-water, limpet-like gastropods. Recent speculation surrounding their affinities has concentrated on their placement within the Gastropoda. However, phylogenetic relationships within the Cocculinoidea, especially the monophyly of families and genera within the group, remain poorly understood. Phylogenetic analysis of 31 morphological characters for 15 cocculinoidean taxa and 2 outgroups resulted in a single most parsimonious tree, length=70, CI=0.62, and RI=0.71. Monophyly of the Cocculinoidea, Cocculinidae, and the genera Cocculina and Coccopigya was supported; Paracocculina and Coccocrater were found to be paraphyletic. Character optimization demonstrates that many characters often cited as diagnostic of various taxa, are often homoplastic and/or synapomorphies at different hierarchical levels.     

Proteinaceous adhesive secretions from insects, and in particular the egg attachment glue of Opodiphthera sp. moths

Biochemical and electrophoretic screening of 29 adhesive secretions from Australian insects identified six types that appeared to consist largely of protein. Most were involved in terrestrial egg attachment. Hydrogel glues were subjected to gravimetric analyses and assessed for overall amino acid composition. When 32 proteins in glues from eight insect species were analyzed individually, many proved to be rich in Gly, Ser, and/or Pro, and some contained substantial levels of 4-hydroxyproline. A few proteins were heavily glycosylated. Abundant protein-based secretions were tested as adhesives, mainly by measuring dry shear strength on wood. The strongest (1-2 MPa) was an egg attachment glue produced by saturniid gum moths of the genus Opodiphthera. It was harvested from female colleterial gland reservoirs as a treacle-like liquid that underwent irreversible gelation, and recovered from the capsules of laid eggs as a highly elastic orange-brown hydrogel that could also display high tack. Its protein-based nature was confirmed and explored by spectroscopy, enzymatic degradation, and 2D gel electrophoresis. Its proteins are mostly 80-95 kDa, and sequences (almost all novel) were established for 23 tryptic peptides. Scanning probe microscopy of Opodiphthera hydrogel in water returned median values of 0.83 nN for adhesion, 63 kPa for modulus, and 87% for resilience. Recombinant mimics of this material might be useful as biodegradable commodity adhesives or as specialty biomedical products.     

A combined approach to the phylogeny of Cephalopoda (Mollusca)

Cephalopoda represents a highly diverse group of molluscs, ranging in habitat from coastal regions to deep benthic waters. While cephalopods remain at the forefront of modern biology, in providing insight into fields such as neurobiology and population genetics, little is known about the relationships within the group. This study provides a comprehensive phylogenetic analysis of Cephalopoda (Mollusca) using a combination of molecular and morphological data. Four loci (three nuclear 18S rRNA, fragments of 28S rRNA and histone H3 and one mitochondrial cytochrome c oxidase subunit I) were combined with 101 morphological characters to test the relationships of 60 species of cephalopods, with emphasis within Decabrachia (squids and cuttlefishes). Individual and combined data sets were analyzed using the direct optimization method, with parsimony as the optimality criterion. Analyses were repeated for 12 different parameter sets accounting for a range of indel/change and transversion/transition cost ratios. Most analyses support the monophyly of Cephalopoda, Nautiloidea, Coleoidea and Decabrachia, however, the monophyly of Octobrachia was refuted due to the lack of support for a Cirroctopoda + Octopoda group. When analyzing all molecular evidence in combination and for total evidence analyses, Vampyromorpha formed the sister group to Decabrachia under the majority of parameters, while morphological data and some individual data sets supported a sister relationship between Vampyromorpha and Octobrachia. Within Decabrachia, a relationship between the sepioids Idiosepiida, Sepiida, Sepiolida and the teuthid Loliginidae was supported. Spirulida fell within the teuthid group in most analyses, further rendering Teuthida paraphyletic. Relationships within Decabrachia and specifically Oegopsida were found to be highly parameter‐dependent. © The Willi Hennig Society 2004.