Functional Organization of the Cardiac Ganglion of the Lobster, Homarus americanus

External recording and stimulation, techniques were used to determine which neurons and interactions are essential for production of the periodic burst discharge in the lobster cardiac ganglion. Burst activity can be modulated by brief single shocks applied to the four small cells, but not by similar stimulation of the five large cells, suggesting that normally one or more small cells primarily determine burst rate and duration. Repetitive electrical stimulation of large cells initiates spike activity in small cells, probably via excitatory synaptic and/or electrotonic connections which may normally act to prolong bursts and decrease burst rate. Transection of the ganglion can result in burst activity in small cells in the partial or complete absence of large cell spike activity, but large cells isolated from small cell excitatory synaptic input by transection or by application of dinitrophenol do not burst. Generally, transections which decrease excitatory feedback to small cells are accompanied by an increase in burst rate, but mean spike frequency over an entire burst cycle stabilizes at the original level within 10–30 min for various groups of cells whose spike-initiating sites are still intact. These and previous results suggest that the system is two layered: one or more small cells generate the burst pattern and impose it on the large cells which are the system's motorneurons.     

Integrative Neurophysiology of the Lobster Cardiac Ganglion

The synchronized bursts of impulses produced by the nine neuronsof the isolated Homarus cardiac ganglion are usually initiatedby Cell 7. Activity in all other cells commences with very shortlatency thereafter. Impulses in most cells originate in triggerzones located 1–2 mm from the cell body, but the firstseveral impulses in Cells 8 and 9 frequently originate in distaltrigger zones some distance from the somata. Large cells fireat a high initial frequency, dropping rapidly to a low frequencyplateau. Small cells exhibit a more tonic behavior and fireat intermediate rates. More anterior small cells tend to firefaster than more posterior ones. The major synaptic interactionsare the impulse-mediated excitatory ones from small cells tolarge cells, and possibly to more anterior small cells. Thereare weak interactions from large cells back onto small cells,and very specific interactions from Cells 1 and 2 onto 3_A, 4_A,5_A, and 3_B 4_B 5_B respectively. The large discrete EPSPs generatedin large cells by small cell impulses appear to be the explanationfor "discrete positioning" in large-cell firing patterns. Inthis situation, large-cell impulses only fire at discrete timesduring the burst, regardless of the actual large-cell pattern. The overall view is of a two-layered neural system in whichthe small cells possess an endogenous oscillatory driver potential,synchronized by synaptic and electrotonic interactions, anddriving a train of impulses in each cell. This activates excitatorysynapses on the large cells, which combined with a triggereddriver potential in each large cell, produces synchronized trainsof motor impulses which activate the heart muscle, causing theheartbeat.     

A Description of a New "Amoebozoan" Isolated from the American Lobster, Homarus americanus

ABSTRACT. Our knowledge of the diversity of amoeboid protists is rapidly expanding as new and old habitats are more fully explored. In 2003, while investigating the cause of an amoeboid disease afflicting lobsters on the East Coast, samples were examined for the presence of amoebae from the carapace washings of the American lobster, Homarus americanus . During this survey a unique community of gymnamoebae was discovered. Among the new taxa discovered was a small Thecamoeba -like organism with a single posteriorly directed pseudopodium. Although resembling Parvamoeba rugata , this amoeba displayed distinctive morphology from that isolate or any other amoebozoan. Phylogenetic analysis shows this amoeba is distantly related to the Thecamoebidae. In this paper we describe the unique morphology of a second species of Parvamoeba and discuss its phylogenetic position with respect to the "Amoebozoa."     

Gross Dissection of the Stomach of the Lobster, Homarus Americanus

The stomach of the American lobster (Homarus americanus) is located in the cephalothorax, between the rostrum and the cervical groove. The anterior end of the stomach is defined by the mouth opening and the posterior end by the bottom of the pylorus. Along the dorsal side of the stomach lies the stomatogastric nervous system (STNS). This nervous system, which contains rhythmic networks that underlie feeding behavior, is an established model system for studying rhythm generating networks and neuromodulation ^1,2. While it is possible to study this system in vivo ³, the STNS continues to produce its rhythmic activity when isolated in vitro. In order to study this system in vitro the stomach must be removed from the animal. This video article describes how the stomach can be dissected from the American lobster. In an accompanying video article⁴ we demonstrate how the STNS can be isolated from the stomach. Open in a separate windowClick here to view.^{(62M, flv)}     

Sub-arctic Populations of European Lobster, Homarus gammarus, in Northern Norway

The European lobster is distributed throughout the south and western regions of the Norwegian coast. A previous lobster allozyme investigation (1993) in the Tysfjord region, north of the Arctic Circle demonstrated that the lobster population from this region was genetically different from lobster samples collected in other parts of Norway. More detailed investigation including supplementary extensive sampling and additional allozyme, microsatellite and mtDNA analyses are reported here. This investigation supports the genetic distinctness of the Tysfjord population and shows that this is mainly due to a reduction (60–70%) in gene diversity (observed heterozygosities and number of alleles) compared with lobsters from more southern regions. In addition to the Tysfjord region, the comprehensive sampling also included lobsters found in the adjacent Nordfolda fjord system. Genetic analyses provided evidence for significant differences between the lobster populations of Tysfjord and Nordfolda, even though they are separated by a coastal distance of only 142 km. The two populations were also different with regards to several biological characteristics such as body size. The genetic difference between these two geographically close populations is likely to be due to the local hydrological conditions, preventing larval dispersal between the fjord systems. Assessment of lobster abundance in the north-west region suggests that the sub-arctic lobster populations are geographically isolated.     

Modulation of Activity of One Neuron by Subthreshold Slow Potentials in Another in Lobster Cardiac Ganglion

A direct demonstration is given of interaction between specific neurons without impulses, via graded slow potentials electrotonically spread from one cell to another. Repetitive polarizing or depolarizing current pulses of 50 to 200 msec. and subthreshold intensity were passed through an intracellular electrode in the soma of a follower cell of the isolated ganglion. When the frequency is near the natural rhythm of impulse bursts corresponding to heart beats and arising in a pacemaker cell 5 to 10 mm. posteriorly, the bursts rapidly become synchronized with the pulses. The effect disappears upon withdrawing the intracellular electrode. Brief pulses or full spikes in the follower are not effective. Hyperpolarizing long pulses attract the burst to a fixed period after the end of the pulse, depolarizations after the beginning of the pulse. The natural rhythm promptly reappears when the pulses are stopped and occasionally breaks through during weak repetitive pulses. Current pulses in postsynaptic cells also alter the threshold of a presynaptic neuron to externally applied stimuli. Some kind of direct, low resistance pathway for electrotonic spread, discriminating against spikes because of their brevity, is inferred, providing a basis for subthreshold interaction which is specific and not by way of a field effect. Due to the sensitivity of modulation of ongoing rhythms, electrotonic currents can be effective even after decrementing over several millimeters.     

Bioaccumulation and Metabolic Effects of the Endocrine Disruptor Methoprene in the Lobster, Homarus americanus

Methoprene is a pesticide that acts as a juvenile hormone agonist.Although developed initially against insects, it has since beenshown to have toxic effects on larval and adult crustaceans.Methoprene was one of several pesticides applied to the WesternLong Island Sound (WLIS) watershed area during the summer of1999; the other pesticides were malathion, resmethrin, and sumethrin.These pesticides were applied as part of a county-by-countyeffort to control the mosquito vector of West Nile Virus. Subsequently,the seasonal lobster catches from the WLIS have decreased dramatically.The lethality of the pesticides to lobsters had been unknown.We studied the effects of methoprene while other investigatorsstudied effects of the other pesticides. We questioned whethermethoprene, through its effects on larvae, adults or both, couldhave contributed to this decline. We found that low levels ofmethoprene had adverse effects on lobster larvae. It was toxicto stage II larvae at 1 ppb. Stage IV larvae were more resistant,but did exhibit significant increases in molt frequency beginningat exposures of 5 ppb. Juvenile lobsters exhibited variationsin tissue susceptibility to methoprene: hepatopancreas appearedto be the most vulnerable, reflected by environmental concentrationsof methoprene inhibiting almost all protein synthesis in thisorgan. Our results indicated that methoprene concentrates in the hepatopancreas,nervous tissue and epidermal cells of the adult lobster. Methoprenealtered the synthesis and incorporation of chitoproteins (cuticleproteins) into adult postmolt lobster explant shells. SDS PAGEanalyses of adult post–molt shell extracts revealed changesin the synthesis of chitoproteins in the methoprene-treatedspecimens, suggesting that methoprene affects the normal pathwayof lobster cuticle synthesis and the quality of the post-moltshell. Although it is likely that a combination of factors ledto the reduced lobster population in WLIS, methoprene may havecontributed both by direct toxic effects and by disrupting homeostaticevents under endocrine control.     

Intrinsic Photosensitive Retinal Ganglion Cells in the Diurnal Rodent,Arvicanthis ansorgei

Intrinsically photosensitive retinal ganglion cells (ipRGCs) represent a new class of photoreceptors which support a variety of non-image forming physiological functions, such as circadian photoentrainment, pupillary light reflex and masking responses to light. In view of the recently proposed role of retinal inputs for the regulation of diurnal and nocturnal behavior, we performed the first deep analysis of the ipRGC system in a diurnal rodent model, Arvicanthisansorgei, and compared the anatomical and physiological properties of ipRGCs with those of nocturnal mice. Based on somata location, stratification pattern and melanopsin expression, we identified two main ipRGC types in the retina of Arvicanthis: M1, constituting 74% of all ipRGCs and non-M1 (consisting mainly of the M2 type) constituting the following 25%. The displaced ipRGCs were rarely encountered. Phenotypical staining patterns of ganglion cell markers showed a preferential expression of Brn3 and neurofilaments in non-M1 ipRGCs. In general, the anatomical properties and molecular phenotyping of ipRGCs in Arvicanthis resemble ipRGCs of the mouse retina, however the percentage of M1 cells is considerably higher in the diurnal animal. Multi-electrode array recordings (MEA) identified in newborn retinas of Arvicanthis three response types of ipRGCs (type I, II and III) which are distinguished by their light sensitivity, response strength, latency and duration. Type I ipRGCs exhibited a high sensitivity to short light flashes and showed, contrary to mouse type I ipRGCs, robust light responses to 10 ms flashes. The morphological, molecular and physiological analysis reveals very few differences between mouse and Arvicanthis ipRGCs. These data imply that the influence of retinal inputs in defining the temporal niche could be related to a stronger cone input into ipRGCs in the cone-rich Arvicanthis retina, and to the higher sensitivity of type I ipRGCs and elevated proportion of M1 cells.     

Some effects of proctolin on the cardiac ganglion of the Maine Lobster, Homarus americanus (Milne Edwards)

The neuropeptide proctolin has excitatory effects on the isolated lobster cardiac ganglion. Selective application to the anterior cell body region produces a dose-dependent (10(-8)--10(-5) M) prolonged depolarization of large anterior cells as well as marked increases in burst frequency and/or duration. In ganglia which have been silenced with tetrodotoxin, proctolin application to anterior cells elicits long-lasting depolarizing responses which are accompanied by a 10-30% increase of the apparent membrane input resistance. Higher proctolin concentrations produce high-frequency trains of driver potentials. It is proposed that a proctolin like peptide may serve a neurohumoral role in the lobster cardiac ganglion and that the anterior motor neurons exhibit endogenous rhythmicity in its presence.     

Structural and Functional Basis of Motor Pattern Generation in the Stomatogastric Ganglion of the Lobster

The stomatogastric ganglion of the lobster Panulirus interruptuscontains about 30 neurons and controls the striated musculatureof the stomach. The ganglion produces two complex rhythms, thepyloric cycle and the gastric mill cycle, when completely deafferented. This paper describes the neural circuitry underlying this activityin terms of interactions among motor neurons. The pyloric motorneurons are coordinated by electrotonic and inhibitory synapticinteractions which are driven by a group of three neurons havingendogenous bursting capability. The gastric mill cycle doesnot appear to have any such driving source, and instead relieson the overall properties of the network to generate its burstpatterns. Preliminary computer modeling indicates that alternatebursting between antagonists can occur without cyclically burstingdriver cells. Computer reconstruction of Procion-filled stomatogastricneurons are used both to corroborate the results of the physiologicalstudies and to quantify the geometry for purposes of modelingthe intraneuronal flow of synaptic currents.     

Phase Maintenance in the Pyloric Pattern of the Lobster (Panulirus interruptus) Stomatogastric Ganglion

The extent to which individual neural networks can producephase-constant motor patterns as cycle frequency is altered has notbeen studied extensively. I investigated this issue in thewell-defined, rhythmic pyloric neural network. When pyloric cyclefrequency is altered three- to fivefold, pyloric inter-neuronaldelays shift by hundreds to thousands of msec, and all pyloricpattern elements show strong phase maintenance. The experimentalparadigm used is unlikely to activate exogenous inputs to thenetwork, and these delay changes are thus likely to arise fromphase-compensatory mechanisms intrinsic to the network. Pyloricinter-neuronal delays depend on the time constants of the networkssynapses and of the membrane properties of its neurons. The observeddelay shifts thus suggest that, in response to changes in overallcycle frequency, these constants vary so as to maintain patternphasing.     

Statistical Estimation of Distribution of Quantal Release of Transmitter in Neuromuscular Synapse of the Lobster Homarus americanus

A new approach to estimation of quantal release distribution of transmitter under conditions of high synaptic activity is presented. Postsynaptic responses of neuromuscular excitatory synapse in muscle-opener of nipper of the lobster, which are obtained by focal extracellular recording, are used as original data set. Based on two data groups (value of evoked and spontaneous postsynaptic responses), the linear regression model is constructed. Parameters of this model describe completely the quantal release distribution. To evaluate the parameters, biased modifications of the least squares method—the penalized least squares method and the principal components method—were applied. As a result, it was possible to achieve estimations of the quantal release distribution with sufficiently low standard errors. Modeling studies have shown that the gain of accuracy of the estimation due to a decrease of the standard error exceeds considerably losses caused by its bias.     

Description of the Blueberry Root-knot Nematode,Meloidogyne carolinensis n. sp.

Meloidogyne carolinensis n. sp. is described from cultivated highbush blueberry (cultivars derived from hybrids of Vaccinium corymbosum L. and V. lamarckii Camp) in North Carolina. The perineal pattern of the female has a large cuticular ridge that surrounds the perivulval area, and the excretory pore is near the level of the base of the stylet. The stylet is 15.9 μm long and the knobs gradually merge with the shaft. The head shape and stylet morphology of the male are quite variable. The typical head and four variants, as well as the typical stylet and two variants, are described. The labial disc, medial lips, and lateral lips of second-stage juveniles are fused and in the same contour. The head region is not annulated. Mean juvenile length is 463.7 μm, stylet length is 11.9 μm, and tail length is 42.5 μm.     

Behavior,Growth and Survival of Stage V Lobsters (Homarus Americanus) in Relation to Shelter Availability and Lobster Density

The effects of 2 levels of shelter availability (8 or 32 shelters) and of 3 levels of intraspecific density (5, 15, or 30 individuals per 0.5 m 2) on the behavior, survival and growth of stage V lobsters over a period of h 25 days were examined through controlled laboratory experiments. Dominance interactions were apparent in all treatments and only certain lobsters gained access to shelter in the medium- and high-density treatments. Lobsters spent more than 95% of their time in shelter when possible, and expressed significantly greater fidelity to one shelter in the 8-shelter treatment compared to the 32-shelter treatment. Shelter availability and lobster density had no effect on the incidence of molting or on the mean size of lobsters at experiment's end. The proportion of injured lobsters was independent of shelter availability, but was positively related to density. Lobsters were most likely to die after having molted and percent mortality was significantly greater at low density and in the 32-shelter treatment than in other treatments, probably due to a stronger and more consistent dominance hierarchy. Overall, the findings suggest that settlers may interact, that the frequency and intensity of interactions may be modified by relative shelter availability and lobster density, and that such interactions may contribute to determine a cohort's fate.     

Behavior, Growth and Survival of Stage V Lobsters ( Homarus Americanus ) in Relation to Shelter Availability and Lobster Density

The effects of 2 levels of shelter availability (8 or 32 shelters) and of 3 levels of intraspecific density (5, 15, or 30 individuals per 0.5 m 2 ) on the behavior, survival and growth of stage V lobsters over a period of ≤25 days were examined through controlled laboratory experiments. Dominance interactions were apparent in all treatments and only certain lobsters gained access to shelter in the medium- and high-density treatments. Lobsters spent more than 95% of their time in shelter when possible, and expressed significantly greater fidelity to one shelter in the 8-shelter treatment compared to the 32-shelter treatment. Shelter availability and lobster density had no effect on the incidence of molting or on the mean size of lobsters at experiment's end. The proportion of injured lobsters was independent of shelter availability, but was positively related to density. Lobsters were most likely to die after having molted and percent mortality was significantly greater at low density and in the 32-shelter treatment than in other treatments, probably due to a stronger and more consistent dominance hierarchy. Overall, the findings suggest that settlers may interact, that the frequency and intensity of interactions may be modified by relative shelter availability and lobster density, and that such interactions may contribute to determine a cohort's fate.     

Description of three new Triaenodes species from Fiji (Trichoptera,Leptoceridae)

The following three new species are illustrated and described from Fiji: Triaenodes rebellus Eriksson & Johanson, sp. n., Triaenodes oscitus Müller & Johanson, sp. n., and Triaenodes forcipatus Puranen Li & Johanson, sp. n. All species are endemic to Viti Levu. A key to the male Triaenodes species of Fiji is provided. With this report, the number of Triaenodes species known from Fiji is doubled.     

Neuropilar Projections of the Anterior Gastric Receptor Neuron in the Stomatogastric Ganglion of the Jonah Crab,Cancer Borealis

Sensory neurons provide important feedback to pattern-generating motor systems. In the crustacean stomatogastric nervous system (STNS), feedback from the anterior gastric receptor (AGR), a muscle receptor neuron, shapes the activity of motor circuits in the stomatogastric ganglion (STG) via polysynaptic pathways involving anterior ganglia. The AGR soma is located in the dorsal ventricular nerve posterior to the STG and it has been thought that its axon passes through the STG without making contacts. Using high-resolution confocal microscopy with dye-filled neurons, we show here that AGR from the crab Cancer borealis also has local projections within the STG and that these projections form candidate contact sites with STG motor neurons or with descending input fibers from other ganglia. We develop and exploit a new masking method that allows us to potentially separate presynaptic and postsynaptic staining of synaptic markers. The AGR processes in the STG show diversity in shape, number of branches and branching structure. The number of AGR projections in the STG ranges from one to three simple to multiply branched processes. The projections come in close contact with gastric motor neurons and descending neurons and may also be electrically coupled to other neurons of the STNS. Thus, in addition to well described long-loop pathways, it is possible that AGR is involved in integration and pattern regulation directly in the STG.     

Multi-Annual Fluctuations in Reconstructed Historical Time-Series of a European Lobster (Homarus gammarus) Population Disappear at Increased Exploitation Levels

Through the history of ecology, fluctuations of populations have been a dominating topic, and endogenous causes of fluctuations and oscillations have been recognized and studied for more than 80 years. Here we analyzed an historical dataset, covering more than 130 years, of European lobster (Homarus gammarus) catches. The data shows periodic fluctuations, which are first dampened and then disappear over time. The disappearance of the periodicity coincided with a substantial increase in fishing effort and the oscillations have not reappeared in the time series. The shifting baseline syndrome has changed our perception of not only the status of the stock, but also the regulating pressures. We describe the transition of a naturally regulated lobster population into a heavily exploited fisheries controlled stock. This is shown by the incorporation of environmental and endogenous processes in generalized additive models, autocorrelation functions and periodicity analyses of time-series.