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Extensive mitochondrial gene arrangements in coleoid Cephalopoda and their phylogenetic implications.

We determined the complete mitochondrial genomes of five cephalopods of the Subclass Coleoidea (Suborder Oegopsida: Watasenia scintillans, Todarodes pacificus, Suborder Myopsida: Sepioteuthis lessoniana, Order Sepiida: Sepia officinalis, and Order Octopoda: Octopus ocellatus) and used them to infer phylogenetic relationships. In our Maximum Likelihood (ML) tree, sepiids (cuttlefish) are at the most basal position of all decapodiformes, and oegopsids and myopsids form a monophyletic clade, thus supporting the traditional classification of the Order Teuthida. We detected extensive gene rearrangements in the mitochondrial genomes of broad cephalopod groups. It is likely that the arrangements of mitochondrial genes in Oegopsida and Sepiida were derived from those of Octopoda, which is thought to be the ancestral order, by entire gene duplication and random gene loss. Oegopsida in particular has undergone long-range gene duplications. We also found that the mitochondrial gene arrangement of Sepioteuthis lessoniana differs from that of Loligo bleekeri, although they belong to the same family. Analysis of both the phylogenetic tree and mitochondrial gene rearrangements of coleoid Cephalopoda suggests that each mitochondrial gene arrangement was acquired after the divergence of each lineage.

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On the depth and scale of metabolic rate variation: scaling of oxygen consumption rates and enzymatic activity in the Class Cephalopoda (Mollusca).

Recent ecological theory depends, for predictive power, on the apparent similarity of metabolic rates within broad taxonomic or functional groups of organisms (e.g. invertebrates or ectotherms). Such metabolic commonality is challenged here, as I demonstrate more than 200-fold variation in metabolic rates independent of body mass and temperature in a single class of animals, the Cephalopoda, over seven orders of magnitude size range. I further demonstrate wide variation in the slopes of metabolic scaling curves. The observed variation in metabolism reflects differential selection among species for locomotory capacity rather than mass or temperature constraints. Such selection is highest among epipelagic squids (Lolignidae and Ommastrephidae) that, as adults, have temperature-corrected metabolic rates higher than mammals of similar size.

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MR microscopy of cobalt-labeled nerve cells and pathways in an invertebrate brain (Sepia officinalis, Cephalopoda).

This article describes a novel application of contrast-enhanced MR microscopy to trace nerve cells and pathways through small invertebrate brains. Using the cuttlefish Sepia officinalis (Cephalopoda) as a model, the cells and pathways of one of the brain nerves were labeled with paramagnetic cobalt(II) ions by conventional centripetal cobalt iontophoresis. In MR microscopy, the cobalt-labeled cell bodies and pathways became hypointense in 9.4 T spin echo images. Their course and distribution were identical with those seen with conventional histological techniques after cobalt sulphide precipitation (with or without subsequent silver intensification). Magn Reson Med 45:575-579, 2001.

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Phylogeny and historical biogeography of the loliginid squids (Mollusca: cephalopoda) based on mitochondrial DNA sequence data.

The cephalopod taxon Loliginidae (Cephalopoda: Myopsida) is a species-rich group of tropical and temperate shallow-water squids, many of which are commercial fisheries objects and neurophysiological research organisms. The worldwide distribution of these squids could make Loliginidae a useful case study in shallow-water marine biogeography, but the phylogeny of the group is unknown. To clarify loliginid phylogeny, regions of two mitochondrial genes (the 16S rRNA and the cytochrome c oxidase subunit I genes) were sequenced for members of 19 loliginid species and several outgroups. Maximum-parsimony and maximum-likelihood analyses were performed on a combined data set, as well as on each data set individually. Analyses of the combined data support loliginid monophyly and reveal four clades-one consisting primarily of species in American waters from two genera, one composed of 3 east Atlantic species, one consisting of the bioluminescent loliginids (Uroteuthis sensu Vecchione et al., 1998) plus Loliolus japonica, and one represented by a Loligo (Alloteuthis) subulata-Lolliguncula mercatoris pair. The likelihood of the unconstrained maximum-likelihood tree is not significantly better than the likelihoods of the best trees constrained to Sepioteuthis monophyly or Uroteuthis monophyly, but there is significant support for Lolliguncula polyphyly. Tests of alternative hypotheses of loliginid cladogenesis suggest that cladogenesis within Loliginidae is correlated with the widening of the Atlantic and the closure of the Tethys Sea, although dispersal from the Indo-West Pacific is a reasonable explanation for the origin of the clade of American loliginines.

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Cytology of the renal appendages of Nautilus (Cephalopoda, Tetrabranchiata).

The renal appendages of tetrabranchiate cephalopods, Nautilus pompilius (L.) and Nautilus macromphalus (G.B. Sow.), were studied using light- and electron-microscopical methods. The appendages, homologous to the renal appendages of dibranchiate cephalopods, possess in both species a folded transporting epithelium characterized by a secreting brush border and a high content of mitochondria and lysosome-like dense bodies as well as a basal labyrinth, and are separated from the blood lacunae by a multilayered lamina basalis. In the extracellular crypt-like infoldings of the epithelium, numerous concentric stratified concrements are found. The results strengthen the hypothesis that these concrements represent mineral-storage structures only found in calcium shell-supporting species within the class Cephalopoda.

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Nitric oxide: a vasodilatatory mediator in the cephalic aorta of Sepia officinalis (L.) (Cephalopoda).

Evidence is presented that nitric oxide (NO) may regulate blood pressure in cephalopod molluscs. In vitro tests performed on the cephalic aorta of Sepia officinalis (L.) (Cephalopoda) showed that the NO releasers (glyceroltrinitrate, sodium nitroprusside, 3-morpholinylsydnoneimine chloride and KNO(2)) induced concentration-dependent vasodilatation of vessel segments (without the tunica adventitia/periadventitia) precontracted by dopamine. These vasodilatatory actions could be totally blocked by oxadiazolo[4,3-a] quinoxalin-1-one, an inhibitor of the NO-sensitive guanylyl cyclase, and partially mimicked by the cyclic guanosine monophosphate (cGMP) analogue 8-bromo cGMP and by the phosphodiesterase inhibitor, zaprinast. The NO-precursor, L-arginine, showed vasodilatatory effects only on segments of the aorta in which the layers containing nerves (tunica adventitia/periadventitia) had been left intact, suggesting that NO synthase may be located within peripheral nerves.

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Evolutionary relationships among squids of the family Gonatidae (Mollusca: Cephalopoda) inferred from three mitochondrial loci.

The oceanic squid family Gonatidae (Mollusca: Cephalopoda) is widely distributed in subpolar and temperate waters, exhibiting behavioral and physiological specializations associated with reproduction. Females of several species undergo muscular degeneration upon maturation; origins of this complex morphogenic change are unknown, hindering our understanding of ecological and morpho-physiological adaptations within the family. To provide further information regarding the evolutionary relationships within Gonatidae, three mitochondrial loci (12S rRNA, 16S rRNA, and cytochrome c oxidase subunit I) were analyzed for 39 individuals representing fourteen gonatid and six outgroup cephalopod species. In addition to elucidating relationships among gonatids, molecular data provided more information than morphological data for problematic specimens. Although some data sets are incongruent or have low nodal support values, combined molecular analysis confirms the presence of gonatid groups previously established by morphological characteristics (i.e., possessing radular teeth in seven longitudinal rows and muscular mantle tissue). These characteristics are basal to taxa possessing radular teeth in five longitudinal rows and less muscular mantle tissue, indicating that the derived forms are those species exhibiting physiological adaptation such as tissue degeneration upon maturation and egg brooding.

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Phylogenetic relationships among cirrate octopods (Mollusca: Cephalopoda) resolved using mitochondrial 16S ribosomal DNA sequences.

PHYLOGENETIC RELATIONSHIPS AMONG THE CIRRATE OCTOPODS (MOLLUSCA: Cephalopoda) were investigated using partial sequences of the 16S rRNA mitochondrial gene. The derived phylogeny supports the traditional separation of cirrate families based on web form. Genera with a single web (Opisthoteuthis, Grimpoteuthis, Luteuthis, and Cirroctopus) are clearly distinct from those with an intermediate or secondary web (Cirroteuthis, Cirrothauma, and Stauroteuthis). The cirrates with a single web are separated into three groups. The first group is represented by Opisthoteuthis species, the second by Grimpoteuthis and Luteuthis, and the third by members of the genus Cirroctopus. There is no support for the isolation of Luteuthis in a separate family (Luteuthidae). There is, however, evidence of two groupings within the genus Opisthoteuthis. The data suggest the following revisions in the systematic classification of the cirrates: (1) Cirrothauma, Cirroteuthis, and Stauroteuthis be united in the Cirroteuthidae; (2) Grimpoteuthis and Luteuthis be placed in the Grimpoteuthidae; (3) Opisthoteuthis in the Opisthoteuthidae, and; (4) Cirroctopus be considered sufficiently distinct from both Opisthoteuthidae and Grimpoteuthidae to warrant placement in a new family.

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From Polyplacophora to Cephalopoda: comparative analysis of nitric oxide signalling in mollusca.

The distribution of putative nitric oxide synthase (NOS)-containing cells has been analysed using NADPH-d histochemistry in the CNS and peripheral tissues in more than 2D ecologically and systematically different molluscan genera representing 3 main classes of the phylum MOLLUSCA: Polyplacophora (Lepidopleurus, Lepidozona, Katharina), Gastropoda (Littorina, Lymnaea, Aplexa, Physa, Planorbarius, Planorbis, Helisoma, Biomphalaria, Helix, Limax, Cepaea, Bulla, Aplysia, Phyllaplysia, Philine, Pleurobranchea, Tritonia, Armina, Flabellina, Cadlina) and Cephalopoda (Octopus, Sepia, Rossia, Loligo). Several species were used for more detailed immunohistochemical, biochemical, biophysical and physiological studies to further assay of NOS activity and to analyse functional roles of nitric oxide (NO) in these animals. The main conclusions of our comparative analysis and literature survey can be summarised as following: (i) There is strong evidence for the presence of NO-dependent signalling pathways in different molluscan species. (ii) We hypothesise that a general tendency in the evolution of NADPH-d-reactive cells in Mollusca is a migration of nitrergic function from periphery to the CNS. Also, different isoforms of NOS appear to be present in any one species. (iii) One of the main functional targets of NO signalling is the feeding system. However, there are obvious differences between predators (many labelled central motoneurons) and herbivorous species (many labelled peripheral putative sensory cells) as well as between land/freshwater and marine animals. Nevertheless, in all species tested NO-activated feeding-like motor patterns in the buccal ganglia. Additional functional and cellular targets for NO in molluscs are also considered. We briefly review neuromodulatory mechanisms of NO action and we consider molluscs as useful model systems for investigations of the roles of NO.

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Distribution and function of biogenic amines in the heart of Nautilus pompilius L. (Cephalopoda, Tetrabranchiata).

Biogenic amines (serotonin and catecholamines), play an important role in the control of the blood flow not only in vertebrates, but also in invertebrates such as cephalopods. In contrast to the well investigated hearts of the a 'modern', coleoid cephalopods, the innervation of the heart of the archaic Nautilus pompilius L. has not been studied in detail. In this study the distribution and effects of biogenic amines in the Nautilus heart were investigated. Serotonin and catecholamines were visualised by the glyxoylic acid induced fluorescence. High performance liquid chromatotography analysis was performed to discriminate between the catecholamines, which showed a high content of noradrenaline in the 4 auricles, the aorta and the ventricle, whereas the ventricle showed a high dopamine content. Adrenaline was found at a very low concentration in the ventricle. Serotonin and dopamine were also immunohistochemically localised to larger nerves and throughout the heart, respectively. In organ bath experiments, the auricles showed little spontaneous activity. After adding serotonin, they displayed rhythmical contractions, which were accelerated dose-dependently by noradrenaline. In summary, these data suggest an important role for biogenic amines in the control of the heart of Nautilus pompilius L., with serotonin possibly stimulating excitatory nerve fibres, whereas noradrenaline is likely to influence the muscle contraction itself.

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Molecular phylogeny of coleoid cephalopods (Mollusca: Cephalopoda) using a multigene approach; the effect of data partitioning on resolving phylogenies in a Bayesian framework.

The resolution of higher level phylogeny of the coleoid cephalopods (octopuses, squids, and cuttlefishes) has been hindered by homoplasy among morphological characters in conjunction with a very poor fossil record. Initial molecular studies, based primarily on small fragments of single mitochondrial genes, have produced little resolution of the deep relationships amongst coleoid cephalopod families. The present study investigated this issue using 3415 base pairs (bp) from three nuclear genes (octopine dehydrogenase, pax-6, and rhodopsin) and three mitochondrial genes (12S rDNA, 16S rDNA, and cytochrome oxidase I) from a total of 35 species (including representatives of each of the higher level taxa). Bayesian analyses were conducted on mitochondrial and nuclear genes separately and also all six genes together. Separate analyses were conducted with the data partitioned by gene, codon/rDNA, gene+codon/rDNA or not partitioned at all. In the majority of analyses partitioning the data by gene+codon was the appropriate model with partitioning by codon the second most selected model. In some instances the topology varied according to the model used. Relatively high posterior probabilities and high levels of congruence were present between the topologies resulting from the analysis of all Octopodiform (octopuses and vampire "squid") taxa for all six genes, and independently for the datasets of mitochondrial and nuclear genes. In contrast, the highest levels of resolution within the Decapodiformes (squids and cuttlefishes) resulted from analysis of nuclear genes alone. Different higher level Decapodiform topologies were obtained through the analysis of only the 1st+2nd codon positions of nuclear genes and of all three codon positions. It is notable that there is strong evidence of saturation among the 3rd codon positions within the Decapodiformes and this may contribute spurious signal. The results suggest that the Decapodiformes may have radiated earlier and/or had faster rates of evolution than the Octopodiformes. The following taxonomic conclusions are drawn from our analyses: (1) the order Octopoda and suborders Cirrata, Incirrata, and Oegopsida are monophyletic groups; (2) the family Spirulidae (Ram's horn squids) are the sister taxon to the family Sepiidae (cuttlefishes); (3) the family Octopodidae, as currently defined, is paraphyletic; (4) the superfamily Argonautoidea are basal within the suborder Incirrata; and (5) the benthic octopus genera Benthoctopus and Enteroctopus are sister taxa.

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