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At least 19 recordsLinked to original sources

Characterization of a G-protein from the mandibular organ of the lobster Homarus americanus (Nephropidae, Decapoda).

1. GTP-binding activity was found in both calf brain and male lobster mandibular organ (MO). There was approximately two to three times as much binding in the calf brain. 2. The GTP-binding activity could be extracted from the calf brain with sodium cholate, but not from the MOs. 3. Using ADP-ribosylation catalyzed by pertussis toxin, GTP-binding was shown to be the result of the presence of G-protein. In the lobster MO the G-protein alpha subunit has a molecular weight of about 42 kDa and may be of the Go or Gi varieties.

Adenosine Diphosphate Ribose↗

Two sniffing strategies in palinurid lobsters.

Most studies of lobster chemoreception have focused on the model systems of Panulirus argus (Palinuridae) and Homarus americanus (Nephropidae). We compare antennule morphology across lobsters and conduct the first kinematic study of antennule flicking in a palinurid species other than P. argus. High-speed video analysis shows that Palinurus elephas flicks at a rate more than an order of magnitude higher than in P. argus. However, both species flick their antennular flagella at a Reynolds number (Re) of approximately one, such that an asymmetry in the speed of the flick phases causes both species to have a leaky closing flick phase and a non-leaky opening phase. The antennular flagella of P. argus are nearly seven times longer than those of P. elephas, and, when compared across palinurid genera, Panulirus species sample far greater areas of water over greater spatial and time scales than do any other palinurid genera. Palinurid lobsters appear to have two sniffing strategies: low flick rates over a large area of water (e.g. P. argus) or high flick rates over a small area of water (e.g. P. elephas). P. argus is a highly informative model system in which to study aquatic chemoreception; however, its antennule anatomy and kinematics suggest a separate strategy, unique to Panulirus species, for sensing chemical plumes in fluid environments.

Animals↗

Hooded sensilla homologues: structural variations of a widely distributed bimodal chemomechanosensillum.

A diversity of sensilla has been described in crustaceans, both across species and within a given species. However, few homologous setal types have been identified in crustaceans. In this study we examined setae with features of the hooded sensillum, which is a class of bimodal chemomechanosensilla first identified on antennules of the Caribbean spiny lobster Panulirus argus. We examined the antennules of 13 species representing seven families of malacostracan crustaceans, and most body surfaces of P. argus, and compared the sensillar morphology from different species and from different body regions to identify interspecific and intraspecific homologues of hooded sensilla. Our results show that sensilla with morphological characteristics of antennular hooded sensilla are present and have a similar pattern of distribution on the antennules of reptantian species representing three families (Palinuridae and Scyllaridae of the Achelata and Nephropidae of the Homarida). Furthermore, hooded sensillar homologues are present on most body surfaces of P. argus. However, there are intraspecific and interspecific variations in the morphology of these sensilla. We present evidence that supports the idea that postembryonic changes in individual sensilla may be responsible for some of these morphological variations. Despite these variations, we conclude that the sensilla are homologues, because they have several common characteristics, similar positions on the body surface, similar substructures, a continuum to their morphological variations, and morphological variation that is correlated with phylogenetic similarity. Taken together these results support the idea that the hooded sensillum is a singular and biologically important sensillar type that has a broad distribution.

Animals↗

Digestion in relation to feeding strategies exhibited by crustacean larvae.

Decapod crustaceans have adopted a full range of reproductive strategies from the release of large numbers of small eggs (Penaeoidea) to the release of relatively low numbers of large advanced larvae (Nephropidae). As larval size determines trophic position in planktonic food webs, all food sources from phyto- to zooplankton are exploited, with many species changing trophic level during ontogenetic development. Comparative studies on digestive enzymes, levels of activity and changes during ontogeny, together with measurements of gastroevacuation rates and food energy values appear to reveal a general pattern. While herbivorous decapod larvae adapt to low food energy values with high enzyme activity levels, rapid food turnover and low assimilation efficiency, carnivorous larvae exhibit low levels of enzyme activity but compensate by extending retention time of high-energy food to maximise assimilation efficiency. New studies on digestive enzyme levels during development in the penaeid Litopenaeus vannamei, the caridean Lysmata debelius and the cirriped Elminius modestus, appear to agree with previous observations.

Animals↗

Developmental expression of the octopamine phenotype in lobsters, Homarus americanus.

We have used immunocytochemical methods to examine the sequence of appearance of octopamine-immunoreactive neurons during development, and to try to correlate that appearance with the emergence of behavioral or physiological capabilities. The first octopamine neurons express their transmitter phenotype at approximately 43% of embryonic development. The last cells show immunostaining at the 3rd larval stage. In the wild, therefore, immunoreactivity in cells appears over a 9-12 month period. In contrast, serotonin-immunoreactive neurons stain early in embryonic development and the last serotonin-immunoreactive cells appear at about the same time the first octopamine-immunoreactive neurons show staining. The pattern of appearance of octopamine-immunoreactive cells is cell type-specific. A pair of brain cells and the descending interneurons stain first. Additional brain cell staining is seen throughout embryonic development. The ascending interneurons appear next, and a general anterior-posterior gradient typifies their emergence over a relatively short portion of embryonic life (E 48-62%). The neurosecretory cell staining appears last, is segment-specific, begins at about 62% development, and continues to the 3rd larval stage. The emergence of immunostaining for amine neurotransmitters within groups of identified neurons at precise times in development may specify possible functional units. With at least one group of cells, this possibility seems plausible: the three pairs of claw octopamine neurosecretory cells show immunostaining as a unit.

Animals↗

Ontogenetic alteration in peptidergic expression within a stable neuronal population in lobster stomatogastric nervous system.

In the adult lobster, Homarus gammarus, the stomatogastric ganglion (STG) contains two well-defined motor pattern generating networks that receive numerous modulatory peptidergic inputs from anterior ganglia. We are studying the appearance of extrinsic peptidergic inputs to these networks during ontogenesis. Neuron counts indicate that as early as 20% of development (E20) the STG neuronal population is quantitatively established. By using immunocytochemical detection of 5-bromo-2'-deoxyuridine incorporation, we found no immunopositive cells in the STG by E70. We concluded that the STG neuronal population remains quantitatively stable from mid-embryonic life until adulthood. We then investigated the ontogeny of FLRFamide- and proctolin-like peptides in the stomatogastric nervous system, from their first appearance until adulthood by using whole mount immunocytochemistry. Numerous FLRFamide-like-immunoreactive STG neuropilar ramifications were observable as early as E45 and remain thereafter. From E50 to the first larval stage, one to three STG somata stained, while somatic staining was not observed in larval stage II and subsequent stages. From E50 and thereafter, the STG neuropilar area was immunopositive for proctolin. One to two proctolinergic somata were detected in the STG of the three larval stages but were not seen in embryos, the post-larval stage or in adults. Thus, peptidergic inputs to the STG are present from mid-embryonic life. Moreover, whereas in the adult, STG neurons only contain glutamate or acetylcholine, some neurons transiently express peptidergic phenotypes during development. Although this system expresses an ontogenetic peptidergic plasticity, the STG neurons produce a single stable embryonic-larval motor output (Casasnovas and Meyrand [1995] J. Neurosci. 15:5703-5718).

Animals↗

Sequential developmental acquisition of cotransmitters in identified sensory neurons of the stomatogastric nervous system of the lobsters, Homarus americanus and Homarus gammarus.

We studied the developmental acquisition of three of the cotransmitters found in the gastropyloric receptor (GPR) neurons of the stomatogastric nervous systems of the lobsters Homarus americanus and Homarus gammarus. By using wholemount immunocytochemistry and confocal microscopy, we examined the distribution of serotonin-like, allatostatin-like, and FLRF(NH2)-like immunoreactivities within the stomatogastric nervous system of embryonic, larval, juvenile, and adult animals. The GPR neurons are peripheral sensory neurons that send proprioceptive information to the stomatogastric and commissural ganglia. In H. americanus, GPR neurons of the adult contain serotonin-like, allatostatin-like, and Phe-Leu-Arg-Phe-amide (FLRF(NH2))-like immunoreactivities. In the stomatogastric ganglion (STG) of the adult H. americanus and H. gammarus, all of the serotonin-like and allatostatin-like immunoreactivity colocalizes in neuropil processes that are derived exclusively from ramifications of the GPR neurons. In both species, FLRF(NH2)-like immunoreactivity was detected in the STG neuropil by 50% of embryonic development (E50). Allatostatin-like immunoreactivity was visible first in the STG at approximately E70-E80. In contrast, serotonin staining was not clearly visible until larval stage I (LI) in H. gammarus and until LII or LIII in H. americanus. These data indicate that there is a sequential acquisition of the cotransmitters of the GPR neurons.

Animals↗

Sequential developmental acquisition of neuromodulatory inputs to a central pattern-generating network.

The activity of the adult stomatogastric ganglion (STG) depends on a large number of aminergic and peptidergic modulatory inputs. Our aim is to understand the role of these modulatory inputs in the development of the central pattern-generating networks of the STG. Therefore, we analyze the developmental and adult expressions of three neuropeptides in the stomatogastric nervous system of the lobsters Homarus americanus and Homarus gammarus by using wholemount immunocytochemistry and confocal microscopy. In adults, red pigment-concentrating hormone (RPCH)-like, proctolin-like, and a tachykinin-like immunoreactivity are present in axonal projections to the STG. At 50% of embryonic development (E50), all three peptides stain the commissural ganglia and brain, but only RPCH- and proctolin-like immunoreactivities stain axonal arbors in the STG. Tachykinin-like immunoreactivity is not apparent in the STG until larval stage II (LII). The RPCH-immunoreactive projection to the STG consists of two pairs of fibers. One pair stains for RPCH immunoreactivity at E50; the second RPCH-immunoreactive pair does not stain until about LII. One pair of the RPCH fibers double labels for tachykinin-like immunoreactivity. The adult complement of neuromodulatory inputs is not fully expressed until close to the developmental time at which major changes in the STG motor patterns occur, suggesting that neuromodulators play a role in the tuning of the central pattern generators during development.

Animals↗

Lobster G-protein coupled receptor kinase that associates with membranes and G(beta) in response to odorants and neurotransmitters.

A cDNA clone (lobGRK2) encoding a protein of 690 amino acids with significant similarity to the GRK2 subfamily of G-protein coupled receptor kinases was isolated. lobGRK2 was widely expressed as a 9-kb major transcript and a protein of 80 kDa. It was most abundant in the brain and the olfactory organ but was absent in the eye/eyestalk. Immunocytochemistry revealed lobGRK2 immunoreactivity in the outer dendritic segments of the olfactory receptor neurons, the site of olfactory transduction. LobGRK2 immunoreactivity was observed in most neuronal structures in the brain, although with varying intensity. It was strongest in neuropil, especially the olfactory and accessory lobes but was also detectable in neuronal cell bodies. Stimulation of brain homogenates with a mixture of neurotransmitters increased the association of lobGRK2 with membranes and with G(beta). Similarly, stimulation of olfactory dendrite homogenates with an odorant mixture caused lobGRK2 to associate with G(beta). These results support the conclusion that lobGRK2 responds to odorants and to neurotransmitters and may act to initiate desensitization by phosphorylating G-protein-coupled receptors in the olfactory organ and the brain, respectively.

Animals↗

Antiport-driven sulfate secretion in an invertebrate epithelium.

A novel invertebrate gastrointestinal transport mechanism has been shown to couple chloride/sulfate exchange in an electrogenic fashion. In the lobster, Homarus americanus, the hepatopancreas, or digestive gland, exists as an outpocketing of the digestive tract, representing a single cell layer separating the gut lumen and an open circulatory system comprised of hemolymph. Investigations utilizing independently prepared brush-border and basolateral membrane vesicles revealed discrete antiport systems which possess the capacity to bring about a transcellular secretion of sulfate. The luminal antiport system functions as a high affinity, one-to-one chloride-sulfate exchanger that is stimulated by an increase in luminal hydrogen ion concentration. Such a system would take advantage of the high chloride concentration of ingested seawater, as well as the high proton concentrations generated during digestion, which further suggests a potential regulation by resident sodium-proton exchangers. Exchange of one chloride for one divalent sulfate ion provides the driving force for electrogenic vectorial translocation. The basolateral antiport system was found to be electroneutral in nature, responsive to gradients of the dicarboxylic anion oxalate, while lacking in proton stimulation. No evidence of sodium-sulfate cotransport, commonly reported for the brush border of vertebrate renal and intestinal epithelia, was observed in either membrane preparation. The two antiporters together can account for the low hemolymph to seawater sulfate levels previously described in decapod crustaceans. A secretory pathway for sulfate based upon electrogenic chloride-antiport may appear among invertebrates partly in response to digestion taking place in a seawater environment.

Animals↗

Immunocytochemical localization of actin and tubulin in the integument of land crab (Gecarcinus lateralis) and lobster (Homarus americanus).

The crustacean integument consists of the exoskeleton and underlying epithelium and associated tissues. The epithelium, which is composed of a single layer of cells, is responsible for the cyclical breakdown and synthesis of the exoskeleton associated with molting (ecdysis). During premolt (proecdysis) the epithelial cells lengthen and secrete the two outermost layers (epicuticle and exocuticle) of the new exoskeleton while partially degrading the two innermost layers (endocuticle and membranous layer) of the overlying old exoskeleton. This increased cellular activity is associated with increased protein synthesis and a change in cell shape from cuboidal to columnar. The cytoskeleton, composed of microfilaments (actin) and microtubules (tubulin), plays important roles in the intracellular organization and motility of eukaryotic cells. Immunoblot analysis shows that the land crab exoskeleton contains actin, tubulin, and actin-related proteins (Varadaraj et al. 1996. Gene 171:177-184). In the present study, immunocytochemistry of land crab and lobster integument showed that both proteins were localized in various cell types, including epithelia, connective tissue, tendinal cells, and blood vessels. Muscle immunostained for actin and myosin, but not for tubulin. The membranous layer of land crab (the other layers of the exoskeleton were not examined) and membranous layer and endocuticle of lobster also reacted specifically with anti-beta-actin and anti-alpha-tubulin monoclonal antibodies, but not with an anti-myosin heavy chain antibody. During proecdysis immunolabeling of the membranous layer decreased probably due to protein degradation. The staining intensity for actin and tubulin in the proecdysial epithelium was similar to that in the intermolt (anecdysial) epithelium, suggesting that there was a net accumulation of both proteins proportional to the increase in cellular volume. These results support the previous biochemical analyses and, more specifically, localize actin and tubulin in exoskeletal structures, suggesting that they may serve both intracellular and extracellular functions in crustaceans. J. Exp. Zool. 286:329-342, 2000.

Actins↗

Lobster GABA receptor subunit expressed in neural tissues.

A cDNA encoding an ionotropic gamma-aminobutyric acid (GABA) receptor subunit was isolated from a lobster (Homarus americanus) cDNA library. A longer version of this cDNA, containing a 108-bp insert, was also detected. The two cDNAs are predicted to encode alternatively spliced proteins of 485 and 521 amino acids, respectively. The sequences were most similar to the Drosophila RDL (resistance to dieldrin) GABA subunit with 54% identity, and 30-35% identity with vertebrate ionotropic GABA receptor subunits. Only the shorter clone formed functional ion channels when transfected into human embryonic kidney (HEK) 293 cells. GABA caused a Cl(-)-selective current in the presence of GABA that was blocked by picrotoxin. The GABA-induced current was weakly sensitive to the GABA(A) antagonist, bicuculline, but was enhanced by pentobarbital. Expression of the GABA receptor mRNA was highest in brain and the olfactory organ, but was not detected in leg muscle. These data suggest that the isolated cDNAs are likely to encode proteins that comprise subunits of native GABA receptors expressed in olfactory receptor neurons and projection neurons of the olfactory deutocerebrum.

Alternative Splicing↗

Glomerular organization in developing olfactory and accessory lobes of American lobsters: stabilization of numbers and increase in size after metamorphosis.

Olfactory glomeruli are columnar and radially arranged at the periphery of the primary chemosensory areas, the olfactory lobes (OLs), in the American lobster Homarus americanus. The number of olfactory glomeruli reaches nearly 100/lobe in midembryonic life, increases rapidly during larval life, and stabilizes at about 200 in juvenile and adult lobsters. The accessory lobes (ALs), higher order integration areas, are composed of cortical columns and of spherical glomeruli. Two populations of spherical glomeruli are defined, the cortical glomeruli located at the bases of the columns, and the medullary glomeruli scattered throughout the ALs. Both cortical columns and spherical glomeruli are seen for the first time in the second larval stage. There are about 1000 cortical columns and 1700 glomeruli/AL in the postlarva and these numbers remain constant during the life of the lobster. In both OLs and ALs, it is the size of the interglomerular spaces and of the glomeruli themselves that increases. Therefore, the data suggest that in both OLs and ALs the glomeruli were already generated when the lobster metamorphoses (stage III to IV) and switches from a planktonic to a benthic existence, and that the new sensory neurons that are formed at each molt in the antennulae grow into existing olfactory glomeruli. Stability of the glomerular population in the primary olfactory centers, once the full complement of glomeruli is acquired, has also been reported in insects, fish, and mammals.

Animals↗

Serotonin depletion by 5,7-dihydroxytryptamine alters deutocerebral development in the lobster, Homarus americanus.

The olfactory and accessory lobes constitute prominent histological structures within the larval and mature lobster deutocerebrum, and both are associated with a dense innervation from paired serotonergic nerve cells, the dorsal giant neurons (DGNs). During development, the cell bodies of the DGNs are the first central somata to express serotonin (5-HT), and the onset of their 5-HT immunoreactivity coincides with the beginning of accessory lobe formation. In contrast, the olfactory lobe anlagen emerge much earlier and grow in the apparent absence of serotonin. The role of serotonergic input for the development of these brain structures was investigated in lobster embryos after serotonin had been depleted pharmacologically with the neurotoxin 5,7-dihydroxytryptamine. A approximately 90% reduction of serotonin was confirmed in eggs using high-performance liquid chromatography with electrochemical detection. Morphometric analyses suggested that serotonin depletion dramatically slowed the growth of olfactory and accessory lobes, although glomeruli differentiated at the normal time in both areas. The toxin exhibited a high degree of specificity for serotonergic neurons and associated target regions, and serotonin depletion persisted for at least 2 months following treatment. The goal of future experiments is to determine which of the cell types that innervate the olfactory and accessory lobes are affected by toxin treatment, thereby resulting in the retarded growth of these areas.

5,7-Dihydroxytryptamine↗