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State-dependent learned valuation drives choice in an invertebrate.

Humans and other vertebrates occasionally show a preference for items remembered to be costly or experienced when the subject was in a poor condition (this is known as a sunk-costs fallacy or state-dependent valuation). Whether these mechanisms shared across vertebrates are the result of convergence toward an adaptive solution or evolutionary relicts reflecting common ancestral traits is unknown. Here we show that state-dependent valuation also occurs in an invertebrate, the desert locust Schistocerca gregaria (Orthoptera: Acrididae). Given the latter's phylogenetic and neurobiological distance from those groups in which the phenomenon was already known, we suggest that state-dependent valuation mechanisms are probably ecologically rational solutions to widespread problems of choice.

Animals↗

Molecular cloning of an invertebrate glutamate receptor subunit expressed in Drosophila muscle.

Insects and other invertebrates use glutamate as a neurotransmitter in the central nervous system and at the neuromuscular junction. A complementary DNA from Drosophila melanogaster, designated DGluR-II, has been isolated that encodes a distant homolog of the cloned mammalian ionotropic glutamate receptor family and is expressed in somatic muscle tissue of Drosophila embryos. Electrophysiological recordings made in Xenopus oocytes that express DGluR-II revealed depolarizing responses to L-glutamate and L-aspartate but low sensitivity to quisqualate, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA), and kainate. The DGluR-II protein may represent a distinct glutamate receptor subtype, which shares its structural design with other members of the ionotropic glutamate receptor family.

Amino Acid Sequence↗

Invertebrate facultative anaerobiosis.

The unique pattern of anaerobic carbohydrate metabolism in invertebrate facultative anaerobes serves to couple other substrate-level phosphorylations to the glycolytic reactions, thus increasing the potential yield of high-energy phosphate compounds. Currently, two important coupling sites can be identified:

Adenosine Triphosphate↗

Associative Learning in Aplysia: evidence for conditioned fear in an invertebrate.

Aversive classical conditioning of Aplysia californica, a gastropod mollusk suited for neurobiological study, produces a learned reaction to the chemosensory conditioned stimulus that is expressed as a marked facilitation of four defensive responses: two graded reflexes (head and siphon withdrawal), an all-or-none fixed act (inking), and a complex fixed action pattern (escape locomotion). In addition, the conditioned stimulus produces a concomitant depression of at least one appetitive response, feeding. These extensive and selective actions of the conditioned stimulus in Aplysia resemble the actions of conditioned fear stimuli in higher mammals and suggest that the functional equivalent of fear occurs in invertebrates and thus may be an adaptive mechanism that is widespread in the animal kingdom.

Animals↗

Assimilation of inorganic nitrogen by marine invertebrates and their chemoautotrophic and methanotrophic symbionts.

Symbioses between marine invertebrates and their chemoautotrophic and methanotrophic symbionts are now known to exist in a variety of habitats where reduced chemical species are present. The utilization of chemical energy and reliance on C(1) compounds by these symbioses are well documented. Much less is known about their metabolism of nitrogen. Earlier work has shown that the tissues of organisms in these associations are depleted of N compared with those of other marine organisms, indicating that local sources of nitrogen are assimilated and that novel mechanisms of nitrogen metabolism may be involved. Although these symbioses have access to rich sources of ammonium (NH(4) and NH(3)) and/or nitrate, several investigators have proposed that N(2) fixation may account for some of these isotope values. Here we report that [N]ammonium and, to a lesser degree, [N]nitrate are assimilated into organic compounds by Solemya reidi, a gutless clam containing S-oxidizing bacteria, and seep mussel Ia, an undescribed mytilid containing methanotrophic bacteria. In contrast, Riftia pachyptila, the giant hydrothermal vent tube worm symbiotic with S-oxidizing bacteria, assimilated nitrate but not exogenous ammonium. The rates of assimilation of these sources are sufficient to at least partially support C(1) compound metabolism. N(2) assimilation was not exhibited by the symbionts tested.

Journal Article↗

alpha-Amylase gene of Streptomyces limosus: nucleotide sequence, expression motifs, and amino acid sequence homology to mammalian and invertebrate alpha-amylases.

The nucleotide sequence of the coding and regulatory regions of the alpha-amylase gene (aml) of Streptomyces limosus was determined. High-resolution S1 mapping was used to locate the 5' end of the transcript and demonstrated that the gene is transcribed from a unique promoter. The predicted amino acid sequence has considerable identity to mammalian and invertebrate alpha-amylases, but not to those of plant, fungal, or eubacterial origin. Consistent with this is the susceptibility of the enzyme to an inhibitor of mammalian alpha-amylases. The amino-terminal sequence of the extracellular enzyme was determined, revealing the presence of a typical signal peptide preceding the mature form of the alpha-amylase.

Amino Acid Sequence↗

Enzyme activities associated with an invertebrate iridovirus: nucleotide phosphohydrolase activity associated with iridescent virus type 6 (CIV).

A nucleoside triphosphate phosphohydrolase activity is firmly associated with a purified invertebrate iridovirus, iridescent virus type 6. The enzyme activity hydrolyzes all the nucleoside triphosphates, but has a high preference for ATP. The products of the reaction are nucleoside diphosphates. Conditions for nucleoside triphosphate phosphohydrolase activity are described.

Adenosine Diphosphate↗

Proteolytic processing of the Sindbis virus membrane protein precursor PE2 is nonessential for growth in vertebrate cells but is required for efficient growth in invertebrate cells.

We have shown previously that processing of the Sindbis virus envelope protein precursor PE2 to envelope protein E2 is not required for virus maturation in cultured vertebrate fibroblast cells and that unprocessed PE2 can be incorporated into infectious virus in place of E2 (J. F. Presley and D. T. Brown, J. Virol. 63:1975-1980, 1989; D. L. Russell, J. M. Dalrymple, and R. E. Johnston, J. Virol. 63:1619-1629, 1989). To better understand the role of this processing event in the invertebrate vector portion of the alphavirus life cycle, we have examined the maturation of Sindbis virus mutants defective in PE2 processing in cultured mosquito cells. We found that although substantial amounts of structural proteins PE2, E1, and C were produced in infected mosquito (aedine) cell lines, very little infectious virus was released. When the period of infection was extended, plaque size variants appeared, some of which exhibited a restored ability to grow in mosquito cells. The nucleotide sequences of two such variants were determined. These variants contained point mutations that restored PE2 cleavage, indicating a genetic linkage between failure to cleave PE2 and failure to grow in mosquito cells.

Aedes↗

Occurrence of Vibrio parahaemolyticus in selected marine invertebrates, sediment, and seawater around Alexandria, Egypt.

Samples of seawater, sediment, and the following invertebrates were collected from the Mediterranean Sea in the neighborhood of Alexandria, Egypt: sea urchins (Echinus spp.), clams (Tapes spp.), and wedge shells (Donax trunculus). A total of 165 samples were collected from November 1979 to July 1980. Average counts (per 100 mL or 100 g) of Vibrio parahaemolyticus content were as follows: seawater, 36; sea urchins, 349; sediment, 436; wedge shells, 534; clams, 1872. Samples collected in the summer months contained higher levels of V. parahaemolyticus than winter samples.

Bivalvia↗

Intracellular calcium translocation during contraction in vertebrate and invertebrate smooth muscles as studied by the pyroantimonate method.

The intracellular localization of activator Ca and its translocation during the mechanical activity were studied on vertebrate and invertebrate smooth muscles by fixing muscle fibers with a 1% OsO4 solution containing 2% potassium pyroantimonate for electron microscopic examination. When guniea-pig taenia coli, Mytilus anterior byssal retractor muscle, and Dorabella longitudinal body wall muscle were fixed during the relaxed state, electron-opaque pyroantimonate precipitate containing Ca was localized along the inner surface of the plasma membrane and at other membranous structures in close apposition to the plasma membrane, in accordance with physiological evidence that these muscles contain intracellularly stored activator Ca. When they were fixed during the contracted state, the precipitate was distributed diffusely in the myoplasm in the form of small particles, indicating the release of activator Ca from the peripheral structures. The contraction in dog coronary artery smooth muscle appears to be associated with the inward movement of extracellular Ca. In accordance with this, the resting coronary artery muscle fibers exhibited the precipitate in the lumen of the caveolae, i.e., the bottle-shaped plasma membrane investigations, but not at the peripheral intracellular structures, though the contracted fibers showed the diffuse distribution of the precipitate in the myoplasm. These results indicate that the pyroantimonate method is very effective in studying the translocation of activator Ca in various types of smooth muscles.

Animals↗

Antibacterial non-glycosidase activity of invertebrate destabilase-lysozyme and of its helical amphipathic peptides.

BACKGROUND: Since bactericidal properties of some lysozymes are independent of their glycosidase activity, we have investigated this phenomenon for destabilase-lysozyme (DL) from medicinal leech (Hirudo medicinalis). METHODS: Glycosidase activity was determined on Micrococcus luteus, non-enzymatic antibacterial activity of heat-treated DL and of synthetic peptides alpha1, alpha2 and alpha3 (fragments of its primary structure) on M. luteus, Escherichia coli, Bacillus brevis and Streptomyces chrysomallus. RESULTS: Glycosidase activity disappeared after the heating of native DL at 100 degrees C for 40 min. Antibacterial activity of heat-treated DL for M. luteus MDMSU128 and MDMSU140 expressed as minimal inhibitory concentration was 9.8.10(-8) and 12.10(-8) M, respectively, and to E. coli MDMSU52 11.10(-8) M. Antibacterial activity of synthetic peptide alpha1 for M. luteus MDMSU128 and for E. coli MDMSU52 was 8.3.10(-5) and 4.9.10(-5) M, respectively. CONCLUSION: DL is the first invertebrate lysozyme with combined enzymatic and non-enzymatic antibacterial action.

Animals↗

A robot trace maker: modeling the fossil evidence of early invertebrate behavior.

The study of trace fossils, the fossilized remains of animal behavior, reveals interesting parallels with recent research in behavior-based robotics. This article reports robot simulations of the meandering foraging trails left by early invertebrates that demonstrate that such trails can be generated by mechanisms similar to those used for robot wall-following. We conclude with the suggestion that the capacity for intelligent behavior shown by many behavior-based robots is similar to that of animals of the late Precambrian and early Cambrian periods approximately 530 to 565 million years ago.

Animals↗

A conserved role for the MEK signalling pathway in neural tissue specification and posteriorisation in the invertebrate chordate, the ascidian Ciona intestinalis.

Ascidians are invertebrate chordates with a larval body plan similar to that of vertebrates. The ascidian larval CNS is divided along the anteroposterior axis into sensory vesicle, neck, visceral ganglion and tail nerve cord. The anterior part of the sensory vesicle comes from the a-line animal blastomeres, whereas the remaining CNS is largely derived from the A-line vegetal blastomeres. We have analysed the role of the Ras/MEK/ERK signalling pathway in the formation of the larval CNS in the ascidian, Ciona intestinalis. We show evidence that this pathway is required, during the cleavage stages, for the acquisition of: (1) neural fates in otherwise epidermal cells (in a-line cells); and (2) the posterior identity of tail nerve cord precursors that otherwise adopt a more anterior neural character (in A-line cells). Altogether, the MEK signalling pathway appears to play evolutionary conserved roles in these processes in ascidians and vertebrates, suggesting that this may represent an ancestral chordate strategy.

Animals↗

Murine Otx1 and Drosophila otd genes share conserved genetic functions required in invertebrate and vertebrate brain development.

Despite the obvious differences in anatomy between invertebrate and vertebrate brains, several genes involved in the development of both brain types belong to the same family and share similarities in expression patterns. Drosophila orthodenticle (otd) and murine Otx genes exemplify this, both in terms of expression patterns and mutant phenotypes. In contrast, sequence comparison of OTD and OTX gene products indicates that homology is restricted to the homeodomain suggesting that protein divergence outside the homeodomain might account for functional differences acquired during brain evolution. In order to gain insight into this possibility, we replaced the murine Otx1 gene with a Drosophila otd cDNA. Strikingly, epilepsy and corticogenesis defects due to the absence of Otx1 were fully rescued in homozygous otd mice. A partial rescue was also observed for the impairments of mesencephalon, eye and lachrymal gland. In contrast, defects of the inner ear were not improved suggesting a vertebrate Otx1-specific function involved in morphogenesis of this structure. Furthermore, otd, like Otx1, was able to cooperate genetically with Otx2 in brain patterning, although with reduced efficiency. These data favour an extended functional conservation between Drosophila otd and murine Otx1 genes and support the idea that conserved genetic functions required in mammalian brain development evolved in a primitive ancestor of both flies and mice.

Animals↗

The structure and physical properties of invertebrate and primitive vertebrate arteries.

Light and electron microscopy and in vitro inflation experiments were conducted on the aortae of three different invertebrate species: the lobster Homarus americanus, the horseshoe crab Limulus polyphemus and the whelk Busycon contrarium. Inflation experiments were also performed on the aortae of two species of primitive vertebrates, the sea lamprey Petromyzon marinus and the Atlantic hagfish Myxine glutinosa. The inflation experiments demonstrated similar overall biomechanical properties in each case, despite the existence of differences in tissue structure. The vessels were compliant at low strains, but demonstrated nonlinear elasticity, increasing in stiffness as strains increased; this property could act as protection against artery wall rupture. The vessels of the lamprey, hagfish and lobster are capable of acting as fairly efficient elastic reservoirs and of smoothing blood flow during circulation as they had low hysteresis values (13-18%). The aortae of the horseshoe crab and whelk, if performing this function, have much higher energy losses, up to more than 30% per cycle. The microscopy studies of the aortae of the lobster, horseshoe crab and whelk revealed tissue structures which differ widely from each other as well as from the structures of the lamprey and hagfish. None of these arteries contained elastin, but all contained fibrillar material which differed in appearance, size and arrangement between species. These materials were conjectured to contribute to the elastic properties of the tissue.

Animals↗

Pathways of inorganic nitrogen assimilation in chemoautotrophic bacteria-marine invertebrate symbioses: expression of host and symbiont glutamine synthetase

Symbioses between chemoautotrophic bacteria and marine invertebrates living at deep-sea hydrothermal vents and other sulfide-rich environments function autotrophically by oxidizing hydrogen sulfide as an energy source and fixing carbon dioxide into organic compounds. For chemoautotrophy to support growth, these symbioses must be capable of inorganic nitrogen assimilation, a process that is not well understood in these or other aquatic symbioses. Pathways of inorganic nitrogen assimilation were investigated in several of these symbioses: the vent tubeworms Riftia pachyptila and Tevnia jerichonana, the vent bivalves Calyptogena magnifica and Bathymodiolus thermophilus, and the coastal bivalve Solemya velum. Nitrate reductase activity was detected in R. pachyptila, T. jerichonana and B. thermophilus, but not in C. magnifica and S. velum. This is evidence for nitrate utilization, either assimilation or respiration, by some vent species and is consistent with the high levels of nitrate availability at vents. The ammonia assimilation enzymes glutamine synthetase (GS) and glutamate dehydrogenase (GDH) were detected in all symbioses tested, indicating that ammonia resulting from nitrate reduction or from environmental uptake can be incorporated into amino acids. A complicating factor is that GS and GDH are potentially of both host and symbiont origin, making it unclear which partner is involved in assimilation. GS, which is considered to be the primary ammonia-assimilating enzyme of autotrophs, was investigated further. Using a combination of molecular and biochemical approaches, host and symbiont GS were distinguished in the intact association. On the basis of Southern hybridizations, immunoreactivity, subunit size and thermal stability, symbiont GS was found to be a prokaryote GS. Host GS was distinct from prokaryote GS. The activities of host and symbiont GS were separated by anion-exchange chromatography and quantified. Virtually all activity in symbiont-containing tissue was due to symbiont GS in R. pachyptila, C. magnifica and B. thermophilus. In contrast, no symbiont GS activity was detected in the gill of S. velum, the predominant activity in this species appearing to be host GS. These findings suggest that ammonia is primarily assimilated by the symbionts in vent symbioses, whereas in S. velum ammonia is first assimilated by the host. The relationship between varying patterns of GS expression and host-symbiont nutritional exchange is discussed.

Journal Article↗

Modulation of the cost of pHi regulation during metabolic depression: a (31)P-NMR study in invertebrate (Sipunculus nudus) isolated muscle.

Extracellular acidosis has been demonstrated to play a key role in the process of metabolic depression under long-term environmental stress, exemplified in the marine invertebrate Sipunculus nudus. These findings led to the hypothesis that acid-base regulation is associated with a visible cost depending on the rate and mode of H(+)-equivalent ion exchange. To test this hypothesis, the effects of different ion-transport inhibitors on the rate of pH recovery during hypercapnia, on energy turnover and on steady-state acid-base variables were studied in isolated body wall musculature of the marine worm Sipunculus nudus under control conditions (pHe 7.90) and during steady-state extracellular acidosis (pHe 7.50 or 7.20) by in vivo (31)P-NMR and oxygen consumption analyses. During acute hypercapnia (2 % CO(2)), recovery of pHi was delayed at pHe 7.5 compared with pHe 7.9. Inhibition of the Na(+)/H(+)-exchanger by 5-(N,N-dimethyl)-amiloride (DMA) at pHe 7.5 delayed recovery even further. This effect was much smaller at pHe 7.9. Inhibition of anion exchange by the addition of the transport inhibitor 4, 4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS) prevented pH recovery at pHe 7.5 and delayed recovery at pHe 7.9, in accordance with an effect on Na(+)-dependent Cl(-)/HCO(3)(-) exchange. The effects of ouabain, DIDS and DMA on metabolic rate were reduced at low pHe, thereby supporting the conclusion that acidosis caused the ATP demand of Na(+)/K(+)-ATPase to fall. This reduction occurred via an inhibiting effect on both Na(+)/H(+)- and Na(+)-dependent Cl(-)/HCO(3)(-) (i.e. Na(+)/H(+)/Cl(-)/HCO(3)(-)) exchange in accordance with a reduction in the ATP demand for acid-base regulation during metabolic depression. Considering the ATP stoichiometries of the two exchangers, metabolic depression may be supported by the predominant use of Na(+)/H(+)/Cl(-)/HCO(3)(-) exchange under conditions of extracellular acidosis.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

Event-related potentials in an invertebrate: crayfish emit 'omitted stimulus potentials'.

Electrical signs of neural activity correlated with stimuli or states include a subclass called event-related potentials. These overlap with, but can often be distinguished from, simple stimulus-bound evoked potentials by their greater dependence on endogenous (internal state) factors. Studied mainly in humans, where they are commonly associated with cognition, they are considered to represent objective signs of moderately high-level brain processing. We tested the hypothesis that invertebrates lack such signs by looking in the crayfish Procambarus clarkii for a class of OFF-effects shown in humans to index expectancy. Disproving the hypothesis, we find, using chronic, implanted preparations, that a good omitted stimulus potential is reliably present. The system learns in a few cycles of a regularly repeated light flash to expect one on schedule. Omitted stimulus potentials are found in the protocerebrum, the circumesophageal connective and in the optic tract - perhaps arising in the retina, as in vertebrates. These potentials can be very local and can include loci with and without direct visual evoked potentials in response to each flash. In some loci, the omitted stimulus potential has a slow wave component, in others only a spike burst. Omitted stimulus potentials are more endogenous than visual evoked potentials, with little dependence on flash or ambient light intensity or on train duration. They vary little in size at different times of the day, but abruptly fail to appear if the ambient light is cut off. They can occur during walking, eating or the maintained defense posture but are diminished by 'distraction' and are often absent from an inert crayfish until it is aroused. We consider this form of apparent expectation of a learned rhythm (a property that makes it 'cognitive' in current usage), to be one of low level, even though some properties suggest endogenous factors. The flashes in a train have an inhibitory effect on a circuit that quickly 'learns' the stimulus interval so that the omitted stimulus potential, ready to happen after the learned interval, is prevented by each flash, until released by a missing stimulus.

Animals↗