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PubMedDate not supplied

Neuromuscular glutamatergic and GABAergic channels.

Our laboratory has worked extensively on glutamatergic and GABA-ergic channels, predominantly in crayfish, but also in locust, Drosophila and recently Ascaris. Channel currents were recorded in the different modes of the patch-clamp technique (Hamill et al., 1981). The opening kinetics of the channels were derived from open and closed time histograms obtained from single channel recordings. From these, channel conductances could also be evaluated. The most relevant data were obtained by very rapidly rising and falling pulses (time of change about 0.1 ms) of agonists applied to outside-out patches containing the respective channels (Franke et al., 1987). From such recordings we constructed dose-response curves for peak and steady-state currents, for the rise times of the currents and for the time constants of desensitization. In double-pulse experiments we measured recovery from desensitization and predesensitization due to low agonist concentrations. For most of the channel types, we succeeded in constructing a reaction scheme which in computer simulations mimicked channel behaviour to a good approximation.

Animals↗
PubMed1998

Characterization of calcium/calmodulin-dependent protein kinase II activity in the nervous system of the lobster, Panulirus interruptus.

Nervous system tissue from Panulirus interruptus has an enzyme activity that behaves like calcium/calmodulin-dependent protein kinase II (CaM KII) This activity phosphorylates known targets of CaM KII, such as synapsin I and autocamtide 3. It is inhibited by a CaM KII-specific autoinhibitory domain peptide. In addition, this lobster brain activity displays calcium-independent activity after autophosphorylation, another characteristic of CaM KII. A cDNA from the lobster nervous system was amplified using polymerase chain reaction. The fragment was cloned and found to be structurally similar to CaM KII. Serum from rabbits immunized with a fusion protein containing part of this sequence immunoprecipitated a CaM KII enzyme activity and a family of phosphoproteins of the appropriate size for CaM KII subunits. Lobster CaM KII activity is found in the brain and stomatogastric nervous system including the commissural ganglia, commissures, stomatogastric ganglion and stomatogastric nerve. Immunoblot analysis of these same regions also identifies bands at an apparent molecular weight characteristic of CaM KII.

Amino Acid Sequence↗
PubMed2000

Decapod crustacean chelipeds: an overview.

The structure, growth, differentiation and function of crustacean chelipeds are reviewed. In many decapod crustaceans growth of chelae is isometric with allometry level reaching unity till the puberty moult. Afterwards the same trend continues in females, while in males there is a marked spurt in the level of allometry accompanied by a sudden increase in the relative size of chelae. Subsequently they are differentiated morphologically into crusher and cutter making them heterochelous and sexually dimorphic. Of the two, the major chela is used during agonistic encounters while the minor is used for prey capture and grooming. Various biotic and abiotic factors exert a negative effect on cheliped growth. The dimorphic growth pattern of chelae can be adversely affected by factors such as parasitic infection and substrate conditions. Display patterns of chelipeds have an important role in agonistic and aggressive interactions. Of the five pairs of pereiopods, the chelae are versatile organs of offence and defence which also make them the most vulnerable for autotomy. Regeneration of the autotomized chelipeds imposes an additional energy demand called "regeneration load" on the incumbent, altering energy allocation for somatic and/or reproductive processes. Partial withdrawal of chelae leading to incomplete exuviation is reported for the first time in the laboratory and field in Macrobrachium species.

Agonistic Behavior↗
PubMed1994

The accumulation and distribution of trace metals in some localized marine species.

Trace elements, such as As, Co, Cr, Hg, Sb, and Zn, were determined by neutron activation analysis (NAA), whereas Cd, Cu, and Pb were determined by graphite furnace atomic absorption spectroscopy (GFAAS) in clam, crab, prawn, swamp cerith, and mussel samples after digestion by microwave heating under controlled conditions before eluting the solutions through a column of a chelating resin, Chelex-100. The standard used in the determination of percentage volatile elements retained by microwave digestion and also in the activation process was Lobster Hepatopancreas TORT-1, whereas known mixed standards were prepared from nitrate salts to determine the efficiency of the separation procedure at a controlled pH. Mercury and lead detected in crabs exceeded the maximum permissible level. Some species also showed a high affinity toward certain elements, and their levels of accumulation in the tissues of these species corresponded with the concentration of these elements in sediments, especially at sites in the vicinity of an industrial zone.

Animals↗
PubMed2000

A central pattern generator to control a pyloric-based system.

A central pattern generator (CPG) is built to control a mechanical device (plant) inspired by the pyloric chamber of the lobster. Conductance-based models are used to construct the neurons of the CPG. The plant has an associated function that measures the amount of food flowing through it per unit of time. We search for the best set of solutions that give a high positive flow of food in the maximization function. The plant is symmetric and the model neurons are identical to avoid any bias in the space of solutions. We find that the solution is not unique and that three neurons are sufficient to produce positive flow. We propose an effective principle for CPGs (effective on-off connectivity) and a few predictions to be corroborated in the pyloric system of the lobster.

Animals↗
PubMed2001

Topology selection by chaotic neurons of a pyloric central pattern generator.

The pyloric Central Pattern Generator (CPG) in the lobster has an architecture in which every neuron receives at least one connection from another member of the CPG. We call this a "non-open" network topology. An "open" topology, where at least one neuron does not receive synapses from any other CPG member, is found neither in the pyloric nor in the gastric mill CPG. Here we investigate a possible reason for this topological structure using the ability to perform a biologically functional task as a measure of the efficacy of the network. When the CPG is composed of model neurons that exhibit regular membrane voltage oscillations, open topologies are as able to maximize this functionality as non-open topologies. When we replace these models by neurons which exhibit chaotic membrane voltage oscillations, the functional criterion selects non-open topologies. As isolated neurons from invertebrate CPGs are known in some cases to undergo chaotic oscillations, this suggests that there is a biological basis for the class of non-open network topologies that we observe.

Animals↗
PubMed2001

Analysis of models for crustacean stretch receptors.

The mechanisms underlying the diverse responses to step current stimuli of models [Edman et al. (1987) J Physiol (Lond) 384: 649-669] of lobster slowly adapting stretch receptor organs (SAO) and fast-adapting stretch receptor organs (FAO) are analyzed. In response to a step current, the models display three distinct types of firing reflecting the level of adaptation to the stimulation. Low-amplitude currents evoke transient firing containing one to several action potentials before the system stabilizes to a resting state. Conversely, high-amplitude stimulations induce a high frequency transient burst that can last several seconds before the model returns to its quiescent state. In the SAO model, the transition between the two regimes is characterized by a sustained pacemaker firing at an intermediate stimulation amplitude. The FAO model does not exhibit such a maintained firing; rather, the duration of the transient firing increases at first with the stimulus intensity, goes through a maximum and then decreases at larger intensities. Both models comprise seven variables representing the membrane potential, the sodium fast activation, fast inactivation, slow inactivation, the potassium fast activation, slow inactivation gating variables, and the intra cellular sodium concentration. To elucidate the mechanisms of the firing adaptations, the seven-variable model for the lobster stretch receptor neuron is first reduced to a three-dimensional system by regrouping variables with similar time scales. More precisely, we substituted the membrane potential V for the sodium fast activation equivalent potential Vm, the potassium fast inactivation Vn for the sodium fast inactivation Vh, and the sodium slow inactivation Vl for the potassium slow inactivation Vr. Comparison of the responses of the reduced models to those of the original models revealed that the main behaviors of the system were preserved in the reduction process. We classified the different types of responses of the reduced SAO and FAO models to constant current stimulation. We analyzed the transient and stationary responses of the reduced models by constructing bifurcation diagrams representing the qualitatively distinct dynamics of the models and the transitions between them. These revealed that (1) the transient firings prior to reaching the stationary state can be accounted for by the sodium slow inactivation evolving more slowly than the other two variables, so that the changes during the transient firings reflect the bifurcations that the two-dimensional system undergoes when the sodium slow inactivation, considered as a parameter, is varied; and (2) the stationary behaviors of the models are captured by the standard bifurcations of a two-dimensional system formed by the membrane potential and the potassium fast inactivation. We found that each type of firing and the transitions between them is due to the interplay between essentially three variables: two fast ones accounting for the action potential generation and the post-discharge refractoriness, and a third slow one representing the adaptation.

Animals↗
PubMed2001

Certification of the methylmercury content in SRM 2977 mussel tissue (organic contaminants and trace elements) and SRM 1566b oyster tissue.

The methylmercury content in two new marine bivalve mollusk tissue Standard Reference Materials (SRMs) has been certified using results of analyses from the National Institute of Standards and Technology (NIST) and two other laboratories. The certified concentrations of methylmercury were established based on the results from four and six different (independent) analytical methods, respectively, for SRM 1566b Oyster Tissue (13.2 +/- 0.7 microg/kg) and SRM 2977 Mussel Tissue (organic contaminants and trace elements) (36.2 +/- 1.7 microg/kg). The certified concentration of methylmercury in SRM 1566b is among the lowest in any certified reference material (CRM).

Animals↗
PubMed2000

Regulatory processes on the cytoplasmic surface of the Na(+)/Ca(2+) exchanger from lobster exoskeletal muscle.

A partially purified preparation of the lobster muscle Na(+)/Ca(2+) exchanger was reconstituted with, presumably, random orientation in liposomes. Ca(2+) efflux from (45)Ca-loaded vesicles was studied in exchanger molecules in which the transporter cytoplasmic surface was exposed to the extravesicular (ev) medium. Extravesicular Na(+) (Na(ev))-dependent Ca(2+) efflux depended directly upon the extravesicular Ca(2+) concentration ([Ca(2+)](ev)) with a half-maximal activation at [Ca(2+)](ev) = 0.6 microm. This suggests that the lobster muscle exchanger is catalytically upregulated by cytoplasmic Ca(2+), as in most other species. In contrast, at low [Na(+)](ev), the Ca(ev)-binding site (i.e., on the cytoplasmic surface) for Ca(2+) transported via Ca(2+)/Ca(2+) exchange was half-maximally activated by about 7.5 microm Ca(2+). Mild proteolysis of the Na(+)/Ca(2+) exchanger by alpha-chymotrypsin also upregulated the Na(ev)-dependent Ca(2+) efflux. Following proteolytic digestion in Ca-free medium, the exchanger was no longer regulated by nontransported ev Ca(2+). Proteolytic digestion in the presence of 1.9 microm free ev Ca(2+), however, induced only a 1. 6-fold augmentation of Ca(2+) efflux, whereas, after digestion in nominally Ca-free medium, a 2.3-fold augmentation was observed; Ca(2+) also inhibited proteolytic degradation of the Na(+)/Ca(2+) exchanger measured by immunoblotting. These data suggest that Ca(2+), bound to a high affinity binding site, protects against the activation of the Na(+)/Ca(2+) exchanger by alpha-chymotrypsin. Additionally, we observed a 6-fold increase in the Na(+)/Ca(2+) exchange rate, on average, when the intra- and extravesicular salt concentrations were increased from 160 to 450 mm, suggesting that the lobster muscle exchanger is optimized for transport at the high salt concentration present in lobster body fluids.

Animals↗
PubMed1997

Gating and conduction properties of a sodium-activated cation channel from lobster olfactory receptor neurons.

The gating and conduction properties of a channel activated by intracellular Na+ were studied by recording unitary currents in inside-out patches excised from lobster olfactory receptor neurons. Channel openings to a single conductance level of 104 pS occurred in bursts. The open probability of the channel increased with increasing concentrations of Na+. At 210 mm Na+, membrane depolarization increased the open probability e-fold per 36.6 mV. The distribution of channel open times could be fit by a single exponential with a time constant of 4.09 msec at -60 mV and 90 mm Na+. The open time constant was not affected by the concentration of Na+, but was increased by membrane depolarization. At 180 mm Na+ and -60 mV, the distribution of channel closed times could be fit by the sum of four exponentials with time constants of 0.20, 1.46, 8.92 and 69.9 msec, respectively. The three longer time constants decreased, while the shortest time constant did not vary with the concentration of Na+. Membrane depolarization decreased all four closed time constants. Burst duration was unaffected by the concentration of Na+, but was increased by membrane depolarization. Permeability for monovalent cations relative to that of Na+ (PX/PNa), calculated from the reversal potential, was: Li+ (1.11) > Na+ (1.0) > K+ (0.54) > Rb+ (0.36) > Cs+ (0.20). Extracellular divalent cations (10 mm) blocked the inward Na+ current at -60 mV according to the following sequence: Mn2+ > Ca2+ > Sr2+ > Mg2+ > Ba2+. Relative permeabilities for divalent cations (PY/PNa) were Ca2+ (39.0) > Mg2+ (34.1) > Mn2+ (15.5) > Ba2+ (13.8) > Na+ (1.0). Both the reversal potential and the conductance determined in divalent cation-free mixtures of Na+ and Cs+ or Li+ were monotonic functions of the mole fraction, suggesting that the channel is a single-ion pore that behaves as a multi-ion pore when the current is carried exclusively by divalent cations. The properties of the channel are consistent with the channel playing a role in odor activation of these primary receptor neurons.

Animals↗
PubMed2000

Comparative toxicity of dissolved metals to early larval stages of Palaemon serratus, Maja squinado, and Homarus gammarus (Crustacea:Decapoda).

The acute lethal toxicities of mercury, copper, and cadmium to the first larval stage of the prawn (Palaemon serratus), spider crab (Maja squinado), and lobster (Homarus gammarus) were tested. Chromium was also tested with the prawn. Mortality was recorded after 48 h (for lobster) or 72 h (for other species) incubation at 18 degrees C, and the median lethal concentrations (LC(50) +/- 95% confidence intervals) per individual and per mass unit were calculated. The LC(50) values were, 74 microg Hg/L, 3,304 microg Cu/L, 1,686 microg Cd/L, 12,486 microg Cr/L for prawn; 72 microg Hg/L, 50 microg Cu/L, 158 microg Cd/L for spider crab; and 48 microg Hg/L, 46 microg Cu/L, 34 microg Cd/L for lobster. Therefore, larvae of spider crab and lobster were markedly more sensitive than prawn to heavy metals and thus more suitable to use in seawater quality bioassays. Of these two species, spider crab is recommended due to its abundance, easier maintenance, and higher fecundity. The mercury LC(50) values for different larval stages of P. serratus (zoea I, II, V, and VI) were obtained, and no ontogenetic change in sensitivity to the metal could be detected.

Animals↗
PubMed1999

Analyses of tissues of eight marine species from Atlantic and Pacific coasts for dioxin-like chlorobiphenyls (CBs) and total CBs.

Eight commercially and recreationally important marine species were collected in 1993 and 1994 from several Atlantic and Pacific coastal regions of the contiguous United States. Approximately 700 edible tissue samples (e.g., whole body of mussel, crustacean muscle and hepatopancreas, and fish muscle) were analyzed for dioxin-like chlorobiphenyls (CBs) and other selected CB congeners using a rapid high-performance liquid chromatography photodiode array detection method (HPLC/PDA). Total CBs and toxic equivalents (TEQs) of dioxin-like CBs were also determined. The most abundant congeners measured in these tissues were the moderately chlorinated CBs (e.g., CB 138, 153), with mean concentrations ranging from below the limits of detection (approximately 0.2 ng/g) to 1,500 ng/g, wet weight. Certain dioxin-like CBs (e.g., CBs 77, 105, 118, 126) were also found in several of these samples (mean concentrations ranging from below the limits of detection [approximately 0.4 ng/g] to 680 ng/g). Similar to previous studies, the majority of seafood tissues contained total CB concentrations that were below the U.S. Food and Drug Administration's (FDA) tolerance limit for CBs of 2,000 ng/g, wet weight (2.0 ppm). Furthermore, the majority of samples that contained CB levels below the FDA CB tolerance limit also had CB TEQs that were lower than the FDA's advisory level for TCDD (25 ppt or 25 pg/g, wet weight) in fish from the Great Lakes, which is used in evaluating CB TEQs. Several crustacean hepatopancreas samples collected from certain Atlantic and Pacific urban sites (e.g., Dungeness crab from Elliott Bay in Puget Sound, WA, American lobster from Deer Island in Boston Harbor, MA), however, did contain total CB and CB TEQs that exceeded the FDA CB tolerance and TCDD advisory limits. Mono-ortho- (e.g., CBs 118, 105) and non-ortho-substituted congeners (e.g., CBs 77 and 126) were the largest contributors to the CB TEQs of the hepatopancreas samples that exceeded the action limit.http://link.springer-ny. com/link/service/journals/00244/bibs/37n2p205.html

Animals↗