Threshold determination for olfactory receptors of the spiny lobster

The shallow water caridean shrimps Heptacarpus pictus and H. paludicola are polymorphic in colour pattern. Populations of these species collected over colour‐variable substrates showed the greatest degree of coloration in terms of the proportion of individuals displaying a distinct colour pattern. The frequencies of H. Pictus colour morphs varied significantly between most sampling periods. Apostatic selection by fish predators is suggested as one hypothesis which could explain these changes in morph frequency.

Experiments with Heptacarpus pictus on rapid colour change showed that, although some pigment migration did occur in the chromatosomes studied, the macroscopic appearance of the colour patterns was not altered when shrimps were shifted from black to white backgrounds or vice versa. The results of background choice experiments with H. pictus suggest that these shrimp do not seek out colour backgrounds that would seem to be a matching background in a concealing coloration. This behaviour is consistent with the morphology of the colour patterns which appears to be a disruptive coloration rather than a concealing coloration that closely matches a particular substrate.     

Serum lipoproteins in the spiny lobster, Panulirus interruptus.

1. Most of the lipids in the hemolymph of the spiny lobster, Panulirus interruptus, were associated with a high density lipoprotein (HDL3). The lipid of this lipoprotein was composed of phospholipid (88%), sterol (4%) and triglyceride (3%). 2. In animals fed 14C-labeled triglyceride radioactivity was not seen in the serum until 12 hr after feeding. Most of this serum radioactivity was associated with phosphatidyl choline. 3. Electron micrographs showed that negatively stained high density lipoproteins of the lobster had a polymorphic appearance.     

Chemotaxis in the Florida spiny lobster, Panulirus argus

The relative importance of input from chemoreceptors on the medial versus the lateral antennular flagellae of the spiny lobster, Panulirus argus, in olfactory orientation is examined. Ablation experiments show that input from the lateral filament, specifically the aesthetase tuft of the lateral filament, is necessary and sufficient to trigger searching behaviour in this organism when stimulated with a gradient of dilute shrimp extract. P. argus locates the odour source by using this lateral filament input in both tropotactic and klinotactic comparisons of odour intensities.     

Mechanisms for mixture suppression in olfactory receptors of the spiny lobster

The mechanisms by which the output of olfactory receptor cellsis suppressed, as can happen, for example, when receptor cellsare activated by stimulus mixtures, are ill defined. We showthat subthreshold concentrations of some odorants suppress theresponses of antennular (olfactory) chemoreceptors of the spinylobster to stimulatory odorants in a manner indicative of competitiveinhibition. The effect of these suppressive odorants on theresponse of other receptor cells is inconsistent with this hypothesis,allowing that non-competitive mechanisms also contribute toperipheral mixture suppression in the olfactory pathway of thespiny lobster.     

Respiratory and calcium transport properties of spiny lobster hepatopancreas mitochondria

Mitochondria were isolated from the hepatopancreas of the Florida spiny lobster Panulirus argus using a high osmolarity medium containing 600 mm mannitol, 83 mm sucrose, 5 mm 4-morpholinepropanesulfonic acid, pH 7.6, 0.5% bovine serum albumin (BSA), and 1 mm EDTA. O₂ uptake and Ca²⁺ transport were measured by electrode methods in similar media (plus 4 mm KP_i, 3.3 mm MgCl₂, and 0.67 mg/ml BSA, with 80 mm KCl replacing a portion of the osmotic support). Substrate-supported respiration was observed to be coupled to phosphorylation of ADP or uptake of Ca²⁺ ions. State 3 rates (nanogram atoms O × minute^?1 × milligram protein^?1 ± SEM (N)) were: 49.2 ± 3.9 (19), succinate; 30.9 ± 3.9 (6), dl-palmitoyl carnitine; 29.0 ± 2.7 (9), l-malate; 40.0 ± 2.3 (3), l-glutamate; 27.7 ± 2.2 (5), d-3-hydroxybutyrate; and 26.4 ± 2.4 (18), l-proline ± pyruvate. α-Glycerol phosphate was not oxidized. Ca²⁺ uptake driven by succinate oxidation proceeded with Ca:O ratios of 4.0 ± 0.2 (SEM). Hepatopancreas mitochondria were not uncoupled by Ca²⁺ uptake in excess of 1100 ng atoms × mg protein^?1. Ca²⁺ efflux could be induced by ruthenium red, indicating the presence of an active Ca²⁺ cycle. These mitochondria may provide a favorable model system in which to study regulation of the Ca²⁺ cycle.     

Polymorphic tetranucleotide microsatellite markers in the Caribbean spiny lobster,Panulirus argus

Ten tetranucleotide microsatellite loci are described for the Caribbean spiny lobster Panulirus argus. Loci were polymorphic (4–15 alleles per locus) and exhibited high levels of expected (0.553–0.921) and observed heterozygosity (0.469–0.906) from samples caught off Belize and Puerto Rico coasts. No significant departure from Hardy–Weinberg equilibrium conditions were observed for any locus. All microsatellite loci should be useful for assessing population discrimination for this valuable marine animal currently subjected to excessive fishing efforts.     

Loss of escape-related giant neurons in a spiny lobster, Panulirus argus

When attacked, many decapod crustaceans perform tailflips, which are triggered by a neural circuit that includes lateral giant interneurons, medial giant interneurons, and fast flexor motor giant neurons (MoGs). Slipper lobsters (Scyllaridae) lack these giant neurons, and it has been hypothesized that behavioral (e.g., digging) and morphological (e.g., flattening and armor) specializations in this group caused the loss of escape-related giant neurons. To test this hypothesis, we examined a species of spiny lobster, Panulirus argus. Spiny lobsters belong to the sister taxon of the scyllarids, but they have a more crayfish-like morphology than scyllarids and were predicted to have escape-related giant neurons. Ventral nerve cords of P. argus were examined using paraffin-embedded sections and cobalt backfills. We found no escape-related giant neurons and no large axon profiles in the dorsal region of the nerve cord of P. argus. Cobalt backfills showed one fewer fast flexor motor neuron than in species with MoGs and none of the fast flexor motor neurons show any of the anatomical specializations of MoGs. This suggests that all palinuran species lack this giant escape circuit, and that the loss of rapid escape behavior preceded, and may have driven, alternative predator avoidance and anti-predator strategies in palinurans.     

Spontaneous and repetitive cardiac slowdown in the freely moving spiny lobster, Panulirus japonicus

The fluctuation of heartbeat interval was investigated to assess cardio-regulatory nervous function in freely moving spiny lobsters. This was performed by time series analysis of the heartbeat interval recorded from restrained animals, freely moving animals, and isolated hearts. The heart rate of freely moving animals exhibited on/off switching: i.e., an elevated and maintained rate was repetitively interrupted by periods of decreased rate. Each period was initiated by a sudden decrease in rate and was terminated by an exponential return to normal activity. In order to explain this characteristic change in heart rate, we have constructed a neurotransmitter release-reuptake model for such bi-stable activity of cardio-regulatory nerves. The model was successful in reproducing the characteristic observed fluctuation. In freely moving animals, the brain seems to regulate the heart through the inhibitory nerve in an "on/off" manner. In the hearts of restrained animals and isolated hearts, the heart rate exhibited white-noise like fluctuation. This implies that stress impairs the normal bi-stable regulatory mode.     

Competition with stone crabs drives juvenile spiny lobster abundance and distribution

Interspecific competition is assumed to have a strong influence on the population dynamics of competing species, but is not easily demonstrated for mobile species in the wild. In the Florida Keys (USA), anecdotal observations have long pointed to an inverse relationship in abundance of two large decapod crustaceans found co-occurring in hard-bottom habitat, the stone crab Menippe mercenaria and the Caribbean spiny lobster Panulirus argus. We used them to explicitly test whether competition for a renewable resource (shelter) can drive the abundance and distribution of the inferior competitor. We first explored this relationship in shelter competition mesocosm experiments to determine the competitively dominant species. Results showed that stone crabs are clearly the dominant competitors regardless of the number of lobsters present, the presence of co-sheltering species such as the spider crab, Damithrax spinosissimus, or the order of introduction of competitors into the mesocosm. We also found that lobsters use chemical cues from stone crabs to detect and avoid them. We then tested the ramifications of this competitive dominance in the field by manipulating stone crab abundance and then tracking the abundance and distribution of spiny lobsters through time. Increased stone crab abundance immediately resulted in decreased lobster abundance and increased aggregation. The opposite occurred on sites where stone crabs were removed. When we stopped removing stone crabs from these sites, they soon returned and lobster abundance decreased. This study explicitly demonstrated that interspecific competition can drive population dynamics between these species, and ultimately, community composition in these shallow water habitats.     

Isolation and characterization of the mitochondrial DNA from the florida spiny lobster, Panulirus argus

1. Mitochondria and mitochondrial DNA were isolated from the digestive gland of Panulirus argus. 2. The mitochondrial DNA has an average contour length of 5.13 microns which corresponds to a molecular weight of 10.10 X 10(6) daltons. 3. The molecular weight based on agarose gel electrophoresis of restricted individual DNA samples ranges from 10.04 to 10.4 X 10(6) daltons. 4. Restriction endonuclease analysis with Bam H1 and Eco R1 demonstrate variation in nucleotide sequence between individual lobsters.     

Reactions of the spiny lobster,Palinunis vulgaris,to substrate tilt (I.)

Summary Movements of legs on a tilting footboard during fore-aft and side-side tilts elicit a number of different reactions in the spiny lobster,Palinurus vulgaris. Most prominent among these are the predominantly phasic equilibrium reactions of the antennae in the direction opposite to footboard tilt, and the compensatory phasic and tonic deviations of the eyestalks in the direction of footboard movement. Systematic movements of the abdomen and uropods also take place, and the legs display resistance reactions which oppose board movement. Stimulation of a single leg is sufficient to produce the major components of these responses. After stiffening of the C-B joints reactions fail to appear. Mechanism and functional significance of these reactions are discussed in the context of the relevant biological stimuli.The support of D.M. Neil by the British Council Younger Research Workers Interchange Scheme is gratefully acknowledged. We would also like to express our thanks to Peter Heinecke for the expert work of developing and factoring the electronic control devices, Eberhard Göldner and Jörg Stadier for the skilful construction of the setup. We are furthermore indebted to Russ Fernald for his help with the computer work.     

Processing of antennular input in the brain of the spiny lobster, Panulirus argus

Neurons in the olfactory deutocerebrum of the spiny lobster, Panulirus argus, were recorded intracellularly and filled with biocytin. Recorded neurons arborized in the olfactory lobe (OL), a glomerular neuropil innervated by olfactory and some presumptive mechanosensory antennular afferents. The neurons responded to chemosensory input from the lateral antennular flagellum bearing the olfactory sensilla but not the medial flagellum bearing many non-olfactory chemosensory sensilla. Many neurons received additional mechanosensory input. Thus the OL integrates specifically olfactory with mechanosensory input. OL neurons had multiglomerular arborizations restricted to one or two of the three horizontal layers of the columnar glomeruli. OL local interneurons comprised core neurons with tree-like neurites and terminals in the base of the glomeruli and rim neurons with neurites surrounding the OL and terminals in the cap/subcap. The somata of OL local interneurons lay in the medial soma cluster (100000 somata). OL projection neurons arborized in the base of the glomeruli and ascended via the olfactory glomerular tract to the lateral protocerebrum. A parallel projection pathway is constituted by projection neurons of the accessory lobe, a glomerular neuropil without afferent innervation but intimate links to the OL. The projection neuron somata constituted the lateral soma cluster (200000 somata).Abbreviations AC anterior cluster (cluster 6,7) - AL accessory lobe - aMC anterior subcluster of medial cluster (cluster 9) - A _lNv main antenna I (antennular) nerve - A _lNM antenna I (antennular) motor nerve - A _llNv main antenna II (antennal) nerve - CB central body - CL central layer of accessory lobe - DC deutocerebral commissure - DCN deutocerebral commissure neuropil - dDUMC dorsal subcluster of dorsal unpaired median cluster (cluster 17) - dMC dorsal subcluster of medial cluster (cluster 11) - dVPALC dorsal subcluster of ventral paired anterolateral cluster (cluster 8) G glomerulus - IDUMC lateral subcluster of dorsal unpaired median cluster (cluster 16) - LC lateral cluster (cluster 10) - LF lateral flagellum of antenna I (antennule) - LL lateral layer of accessory lobe - MF medial flagellum of antenna I (antennule) - ML medial layer of accessory lobe - MPN anterior and posterior median protocerebral neuropils - OGT olfactory globular tract - OGTN olfactory globular tract neuropil - OL olfactory lobe - OLALT olfactory lobe-accessory lobe tract - PB protocerebral bridge - pMC posterior subcluster of medial cluster (cluster 9) - PT protocerebral tract - TNv tegumentary nerve - VPMC ventral paired medial cluster (cluster 12) - VUMC ventral unpaired medial cluster (cluster 13) - vVPALC ventral subcluster of ventral paired anterolateral cluster (cluster 8) - ASW artificial sea water - M3 mixture 3 - PRO L-proline - TM TetraMarin extract     

Hemocyanin-derived phenoloxidase activity in the spiny lobster Panulirus argus (Latreille, 1804)

Hemocyanin and phenoloxidase belong to the type-3 copper protein family, sharing a similar active center whereas performing different roles. In this study, we demonstrated that purified hemocyanin (450 kDa) from the spiny lobster Panulirus argus shows phenoloxidase activity in vitro after treatment with trypsin, chymotrypsin and SDS (0.1% optimal concentration), but it is not activated by sodium perchlorate or isopropanol. The optimal pHs of the SDS-activated hemocyanin were 5.5 and 7.0. Hemocyanin from spiny lobster behaves as a catecholoxidase. Kinetic characterization using dopamine, L-DOPA and catechol shows that dopamine is the most specific substrate. Catechol and dopamine produced substrate inhibition above 16 and 2 mM respectively. Mechanism-based inhibition was also evidenced for the three substrates, being less significant for L-DOPA. SDS-activated phenoloxidase activity is produced by the hexameric hemocyanin. Zymographic analysis demonstrated that incubation of native hemocyanin with trypsin and chymotrypsin, produced bands of 170 and 190 kDa respectively, with intense phenoloxidase activity. Three polypeptide chains of 77, 80 and 89 kDa of hemocyanin monomers were identified by SDS-PAGE. Monomers did not show phenoloxidase activity induced by SDS or partial proteolysis.     

Morphology and physiology of multiglomerular olfactory projection neurons in the spiny lobster

Whole-cell patch-clamp recording was used to characterize olfactory projection neurons in an isolated brain preparation of the spiny lobster, Panulirus argus. Responses to electrical stimulation of the olfactory afferents were recorded from projection neuron somata using biocytin-filled electrodes. All projection neurons were multiglomerular, innervating up to 80% of all olfactory lobe glomeruli, but the innervation was heterogeneous. Most neurons densely innervated only 3–4 glomeruli; the remaining glomeruli in their dendritic arbor were sparsely innervated, thereby creating two distinct patterns of intraglomerular branching. Projection neurons responded to orthodromic stimulation with an initial depolarization and spiking followed by a 1–3 s hyperpolarization. The inhibitory phase of the response was lower in threshold and longer in latency than the excitatory phase, a response pattern also reported in olfactory projection neurons of insects and vertebrates. The somata of the projection neurons supported voltage-activated currents and TTX-sensitive action potentials, suggesting that the soma, although spatially separated from the axon and dendrites, may play a significant functional role in these cells. Dye coupling between some projection neurons correlated with the presence of multiple amplitude action potentials, suggesting that at least some projection neurons may be coupled via gap junctions.