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PubMed1995

Mollusk genes encoding lysine tRNA (UUU) contain introns.

New intron-containing genes encoding tRNAs were discovered when genomic DNA isolated from various animal species was amplified by the polymerase chain reaction (PCR) with primers based on sequences of rabbit tRNA(Lys). From sequencing analysis of the products of PCR, we found that introns are present in several genes encoding tRNA(Lys) in mollusks, such as Loligo bleekeri (squid) and Octopus vulgaris (octopus). These introns were specific to genes encoding tRNA(Lys)(CUU) and were not present in genes encoding tRNA(Lys)(CUU). In addition, the sequences of the introns were different from one another. To confirm the results of our initial experiments, we isolated and sequenced genes encoding tRNA(Lys)(CUU) and tRNA(Lys)(UUU). The gene for tRNA(Lys)(UUU) from squid contained an intron, whose sequence was the same as that identified by PCR, and the gene formed a cluster with a corresponding pseudogene. Several DNA regions of 2.1 kb containing this cluster appeared to be tandemly arrayed in the squid genome. By contrast, the gene encoding tRNA(Lys)(CUU) did not contain an intron, as shown also by PCR. The tRNA(Lys)(UUU) that corresponded to the analyzed gene was isolated and characterized. The present study provides the first example of an intron-containing gene encoding a tRNA in mollusks and suggests the universality of introns in such genes in higher eukaryotes.

Animals↗
PubMed1991

Separation of nucleotides in homogenates of octopus retina by ion-pair reversed-phase liquid chromatography and identification by mass spectrometry.

An isocratic ion-pair reversed-phase liquid chromatographic method has been developed for the determination of thirteen nucleotides including cyclic AMP and cyclic GMP. The resolution capability of this method was evaluated successfully using homogenates of octopus retina, the aim being to elucidate the role of nucleotides (particularly ADP and ATP) in the control of oxidative metabolism. To overcome the inherent lack of specificity of ultraviolet detection we used the coupling of liquid chromatography with mass spectrometry, via a thermospray interface, to confirm the identity of the nucleotides of interest in the biological samples.

Animals↗
PubMedDate not supplied

Marine envenomations. Part 2: Invertebrates.

As more people travel to the oceans for sport diving and other marine-related activities, the incidence of marine envenomations has risen. This article is designed to give the emergency medicine physician an overview of varying marine envenomations, their clinical presentation, and recommended treatment. Part 1 of this article dealt with general wound management and vertebrate envenomations. Part 2 addresses invertebrate envenomations.

Animals↗
PubMedDate not supplied

Toxic exposure to ethylene dibromide and mercuric chloride: effects on laboratory-reared octopuses.

The effects of acute and chronic exposure to either ethylene dibromide (EDB) or mercuric chloride (MC) were studied in laboratory-reared Octopus joubini, O. maya and O. bimaculoides. The advantages of using octopuses were that the responses were immediate, highly visible and sensitive. All species demonstrated signs of toxicity to acute and chronic exposure to EDB and to MC. A dosage-sensitive relationship for the loss and subsequent recovery of locomotor response and of chromatophore expansion was found for each species after acute exposure. For each species the LC50 for chronic exposure occurred within 12 hr at 100 mg/l for EDB and within 3 hr at 1,000 mg/l for MC. This study demonstrated the potential usefulness of laboratory-reared octopuses in evaluating the toxicity of marine environmental pollutants.

Animals↗
PubMed1996

Spin-labeling analysis of structure and dynamics in octopus rhodopsin.

The location of cysteines accessible in octopus rhodopsin were characterized by a spin-labeling technique. Two cysteines were found to bind a methanthiosulfonate spin label. One of the spin labels is attached to helix V with the side chain located within the membrane, most probably close to the polar head group region. The second spin label was found to be attached to cysteine 345 in the C terminus. Light-induced reversible electron paramagnetic resonance spectral changes were observed for the spin label attached at position 345. It is concluded that conformational changes occur during the rhodopsin to metarhodopsin transition in the vicinity of the C-terminus position 345.

Animals↗
PubMed2006

Octopus arm choice is strongly influenced by eye use.

This study aims to investigate the octopus' eye and arm coordination and raises the question if visual guidance determines choice of arm use. Octopuses possess eight seemingly identical arms but have recently been reported to show a preference as to which arm they use to initiate contact with objects. These animals also exhibit lateralized eye use, therefore, a connection between eye and arm preference seems possible. Seven Octopus vulgaris were observed during approach, contact initiation and exploration of plastic objects that were positioned on three different levels in the water column. The subjects most commonly used an arm to initiate contact with an object that was in a direct line between the eye used to look at the object, and the object itself. This indicates that choice of arm use is spatially rather opportunistic when depending on visual guidance. Additionally, first contact with an object was usually established by the central third of the arm and in arm contact sequences neighboring arms were the most likely to follow an arm already touching the object. Although results point towards strong eye/arm coordination, we did not find lateralized behavior in this experiment. Results are discussed from a neuro-anatomical, behavioral and ecological perspective.

Animals↗
PubMed2006

Brain and behavioural evidence for rest-activity cycles in Octopus vulgaris.

Octopus vulgaris maintained under a 12/12h light/dark cycle exhibit a pronounced nocturnal activity pattern. Animals deprived of rest during the light period show a marked 'rebound' in activity in the following 24h. 'Active' octopuses attack faster than 'quiet' animals and brain activity recorded electrically intensifies during 'quiet' behaviour. Thus, in Octopus as in vertebrates, brain areas involved in memory or 'higher' processes exhibit 'off-line' activity during rest periods.

Animals↗
PubMed2003

Single exon structures of the oxytocin/vasopressin superfamily peptides of octopus.

Two genes of the oxytocin/vasopressin superfamily in the cephalopod Octopus vulgaris, cephalotocin (CT) and octopressin (OP), lack introns and consist of a single exon in their protein-coding regions, which is unlike the 2 intron-3 exon structures in most vertebrates and Lys-conopressin in a pond snail Lymnaea stagnalis. Octopus may have lost introns during the evolutionary process in mollusks. mRNA that had deleted 202 bp from preproCT cDNA (CT-del) was also expressed in the brain. The deletion occurred in the central part of neurophysin. Immunohistochemical studies suggest that a translational product of CT-del mRNA may not be present in a stable form due to the loss of neurophysin. Genomic Southern blot analysis revealed that a single copy of each of OP-, CT-, and CT-del-genes was present in the genome.

Amino Acid Sequence↗
PubMed2006

Calretinin distribution in the octopus brain: an immunohistochemical and in situ hybridization histochemical analysis.

The distribution of calretinin containing neurons examined by in situ hybridization mapping was compared with that obtained by immunocytochemistry in the brain of octopus. Results revealed a close correspondence between the two types of investigations. Western blot analysis disclosed a 29 kDa protein immunostained with anti-calretinin antibody. Calretinin containing neurons were localized mainly in the cortex of octopus lobes, including the vertical, frontal, basal, buccal, palliovisceral, pedal and branchial, with variations of staining intensity and density of immunoreactive cells. The amacrine cells surrounding calretinin containing neuronal bodies of the cortex were also labeled unlike the glial cells. The close correspondence of blotting analysis, immunocytochemistry and in situ hybridization indicates with no doubt that calretinin, like other calcium-binding proteins previously studied, is also present in the nervous system of cephalopods. Furthermore, although recent findings localize calretinin also in endocrine glands, the presence of this calcium-binding protein in the brain of octopus indicates that calretinin appeared early in the phylogeny as a neuronal protein already in invertebrates.

Amacrine Cells↗
PubMed2004

Biochemical dynamics of spermatogenesis and oogenesis in Eledone cirrhosa and Eledone moschata (Cephalopoda: Octopoda).

The effect of spermatogenesis and oogenesis on protein, lipid, glycogen, cholesterol and energy contents, total amino acid and fatty acid profiles of Eledone cirrhosa and Eledone moschata tissues (gonad, digestive gland and muscle) was investigated. A significant (p<0.05) increase in the amino acids and protein content of the gonad throughout sexual maturation (namely in oogenesis) was observed, but the allocation of these nitrogen compounds from the digestive gland and muscle was not evident. The major essential amino acids (EAA) in the three tissues were leucine, lysine and arginine. The major nonessential amino acids (NEAA) were glutamic acid, aspartic acid and alanine. A significant increase in lipid and fatty acid contents of gonad and digestive gland was observed. There was also little evidence of accumulated lipid storage reserves being used for egg production. It seems that for egg production Eledone species use energy directly from food, rather than from stored products. Most of saturated fatty acid (SFA) content of the three tissues was presented as 16:0 and 18:0, monounsaturated fatty acid (MUFA) content as 18:1 and 20:1 and polyunsaturated fatty acid (PUFA) content as 20:4n-6, 20:5n-3 and 22:6n-3. Cholesterol and glycogen contents significantly increased in gonad and digestive gland throughout maturation while the muscle revealed no obvious pattern. If Eledone's component sterols are of a dietary origin, a considerable variation in the cholesterol content between species might be expected on the basis of the sterol composition of their prey. Although spermatogenesis and oogenesis had a significant effect (p<0.05) in gonad and digestive gland energy content, the biochemical composition of digestive gland and muscle may not be primarily influenced by sexual maturation, but rather by other biotic factors such as feeding activity, food availability, spawning and brooding.

Animals↗
PubMed2004

Cloning and characterization of an mRNA encoding a novel G protein alpha-subunit abundant in mantle and gill of pearl oyster Pinctada fucata.

Nacre formation is an ideal model to study biomineralization processes. Although much has been done about biomineralization mechanism of nacre, little is known as to how cellular signaling regulates this process. We are interested in whether G protein signaling plays a role in mineralization. Degenerate primers against conserved amino acid regions of G proteins were employed to amplify cDNA from the pearl oyster Pinctada fucata. As a result, the cDNA encoding a novel G(s)alpha (pfG(s)alpha) from the pearl oyster was isolated. The G(s)alpha cDNA encodes a polypeptide of 377 amino acid residues, which shares high similarity to the octopus (Octopus vulgaris) G(s)alpha. The well-conserved A, C, G (switch I), switch II functional domains and the carboxyl terminus that is a critical site for interaction with receptors are completely identical to those from other mollusks. However, pfG(s)alpha has a unique amino acid sequence, which encodes switch III and interaction sites of adenylyl cyclase respectively. In situ hybridization and Northern blotting analysis revealed that the oyster G(s)alpha mRNA is widely expressed in a variety of tissues, with highest levels in the outer fold of mantle and epithelia of gill, the regions essential for biomineralization. We also show that overexpression of the pfG(s)alpha in mammalian MC3T3-E1 cells resulted in increased cAMP levels. Mutant pfG(s)alpha that has impaired CTX substrate diminished its ability to induce cAMP production. Furthermore, the alkaline phosphatase (ALP) activity, an indicator for mineralization, is induced by the G(s)alpha in MC3T3-E1 cells. These results indicated that G(s)alpha may be involved in regulation of physiological function, particularly in biological biomineralization.

Alkaline Phosphatase↗
PubMed2005

Interannual patterns of variation in concentrations of trace elements in arms of Octopus vulgaris.

Concentrations of essential (copper, iron, manganese, selenium, and zinc) and non-essential (arsenic, cadmium, mercury, and lead) trace elements were measured in arms of Octopus vulgaris. The cephalopods were sampled from commercial fishery landings at two sites on the Portuguese coast in spring 2002 and 2003. Mercury was determined using an Advanced Mercury Analyser Spectrophotometer (AMAS) and other trace elements were measured using inductively coupled plasma-atomic emission spectrometry (ICP-AES). Since high levels of arsenic were detected, identification of the forms present was carried out using high performance liquid chromatography (HPLC) followed by ICP-MS. Mean concentrations of trace elements analysed were in the following order: As > Zn > Fe > Cu >> Cd > Pb > Se > Mn >> Hg. The results of speciation of arsenic demonstrate that virtually all arsenic was in the arsenobetaine form, which is the less toxic form. The concentrations of several trace elements were generally high in samples from Viana in 2002. Cadmium concentrations were above the legal limit for human consumption in samples from Viana in 2002 and two of these animals also had lead concentrations that exceeded legal limits. Mercury appeared in all samples but levels were within legally defined safe limits. No relationship was detected between trace element concentrations and size or maturity of octopus.

Analysis of Variance↗
PubMed2005

Concentrations and composition of organochlorine contaminants in different species of cephalopod molluscs from the Italian waters (Adriatic Sea).

Persistent organochlorines, such as polychlorinated biphenyls including coplanar congeners and DDT compounds were measured in the liver of different species of mollusc cephalopods: broadtail squid (Illex coindetii), horned octopus (Eledone moschata) and spider octopus (Octopus salutii) collected in two different areas of the Adriatic Sea (central and southern Adriatic Sea). The concentration of PCBs exceeded DDT ones in horned octopus (PCBs: 617.0 ngg(-1) lipid weight, DDTs: 188.0 ngg(-1) lipid weight) and in spider octopus (PCBs: 590.0 ngg(-1) lipid weight, DDTs: 293.0 ngg(-1) lipid weight), while in broadtail squid PCB load (748.0 ngg(-1) lipid weight) was similar to those of DDTs (514.5 ngg(-1) lipid weight). No significant difference in PCB levels was observed among species, while DDT concentrations were higher in broadtail squid than in horned octopus and spider octopus. DDTs composition showed slight differences in relation to species, thought p,p'-DDE was the predominant compound in all cephalopod species (broadtail squid: 88.1%, horned octopus 94.1%, spider octopus: 97.5%). PCB isomer profiles were similar among species being hexachlorinated isomers the most abundant, followed by penta- and heptachlorobiphenyls, while tri- and tetrachlorobiphenyls made up a small percentage of the total PCB residues. Regarding coplanar congeners, non-ortho PCBs were below the detection limit in all samples and the 2,3,7,8-TCDD toxic equivalent (TEQ) concentrations were very low. The influence of biological and ecological factors (size/weight and location) on the bioaccumulation of organochlorine compounds has been discussed.

Animals↗
PubMed2006

Molecular characterization of the developmental gene in eyes: through data-mining on integrated transcriptome databases.

OBJECTIVES: Our aim was to utilize publicly available and proprietary sources to discover candidate genes important for ocular development. DESIGN AND METHODS: The collated information on our 5092 non-redundant clusters was grouped and functional annotation was conducted using gene ontology (FatiGO) for categorizing them with respect to molecular function. The web-based viewer technological platform (H-InvDB) was employed for transcription analyses of in-house high quality fetal eye Expressed Sequence Tags (ESTs). Eye-specific ESTs were also analyzed across species by using EMBEST. RESULTS: According to adult eye cDNA libraries, nucleic acid binding and cell structure/cytoskeletal protein genes were the most abundant among the ESTs of fetal eyes. Using cDNA assembly in H-InvDB, 20 (80%) of the 25 most commonly expressed genes in the human eye are also expressed in extraocular tissues. The crystalline gamma S gene is highly expressed in the eye, but not in other tissues. We used EMBEST to compare human fetal eye and octopus eye ESTs and the expression similarity was low (1.6%). This indicated that our fetal eye library contains genes necessary for the developmental process and biological function of the eye, which may not be expressed in the fully developed octopus eyes. The human fetal eye cDNA library also contained highly abundant eye tissue genes, including alphaA-crystallin, eukaryotic translation elongation factor 1 alpha 1 (EEF1A1), bestrophin (VMD2), cystatin C, and transforming growth factor, beta-induced (BIGH3). CONCLUSIONS: Our annotated EST set provides a valuable resource for gene discovery and functional genomic analysis. This display will help to appreciate the strengths and weaknesses of the different technological platforms, so that in future studies the maximum amount of beneficial information can be derived from the appropriate use of each method.

Animals↗
PubMed2005

Identifying hallmarks of consciousness in non-mammalian species.

Most early studies of consciousness have focused on human subjects. This is understandable, given that humans are capable of reporting accurately the events they experience through language or by way of other kinds of voluntary response. As researchers turn their attention to other animals, "accurate report" methodologies become increasingly difficult to apply. Alternative strategies for amassing evidence for consciousness in non-human species include searching for evolutionary homologies in anatomical substrates and measurement of physiological correlates of conscious states. In addition, creative means must be developed for eliciting behaviors consistent with consciousness. In this paper, we explore whether necessary conditions for consciousness can be established for species as disparate as birds and cephalopods. We conclude that a strong case can be made for avian species and that the case for cephalopods remains open. Nonetheless, a consistent effort should yield new means for interpreting animal behavior.

Animals↗
PubMed2006

[The modalities of the sexual cycle of Octopus vulgaris in the southern Moroccan Atlantic (Tantan, Boujdour)].

The reproduction of Octopus vulgaris between Tan Tan and Boujdour is studied. The samples used are sorted monthly from commercial catches of coastal trawlers operating in this area from December 2001 to May 2003. The study shows that sexual maturity is in advance for males than for females. Two spawning periods are made out by the follow-up of the RGS: a spring period from March to July, and an autumnal period, which is less intense, between September and October. The division of spermatogonia and spermatocytes reaches its maximal intensity in July and December, whereas the spermiogenesis is active until the time of mating. The vitellogenesis starts in mid-December and extends until the following spawning period. This reveals a later gonadic development for females than for males; approximately seven months for females and three months for males.

Animals↗