Descripción: Split lamellae of posterior gills of Chasmagnathus granulatus adapted to 2.5‰ salinity were mounted in a modified Ussing chamber. With NaCl-saline on both sides of the preparation a transepithelial voltage (Vte) of 4.1±0.5 mV (outside positive) was measured. After voltage-clamping, the negative short-circuit current (Isc) amounted to -142±21 μA cm-2 at a conductance (Gte) of 44±5 mS cm-2. Substitution of either chloride (by nitrate) or sodium (by choline) on both sides of split gill lamellae significantly reduced Isc (by 70-80%) and Gte (by 30-50%). External CsCl (but not BaCl2 or furosemide) inhibited the negative Isc without affecting Gte. Addition of ouabain, BaCl2 or diphenylamine-2-carboxylate to the internal bath inhibited Isc at unchanged Gte. Internal acetazolamide did not affect Isc or Gte across split gill lamellae. Unidirectional Na+ influx across isolated and perfused posterior gills, however, was reduced by internal acetazolamide by approximately 20% at constant Vte. The results suggest that posterior gills of hyperosmoregulating C. granulatus display a high conductance epithelium that actively absorbs NaCl in a coupled way by an electrogenic mechanism similar to that seen in the thick ascending limb of Henle's loop and, to a minor degree, by an electroneutral mechanism, presumably via apical Na+/H+and Cl-/HCO3--antiports.

Descripción: Coupling experiments at small spatial scales with large-scale surveys can help to generalize experimental results across large spatial scales. The goal of the present study was to evaluate patterns of crab herbivory within and, at a larger scale, between many southwestern (SW) Atlantic salt marshes. Hence, we conducted experiments in an Argentinean salt marsh to elucidate the effects of crab Chasmagnathus granulatus grazing on the cordgrass Spartina densiflora and the factors that can affect this interaction, and then examined the potential generality of these results across SW Atlantic salt marshes from Brazil to Argentina (15 marshes, range ≈ 2000 km). Experimental examination of the effects of crabs (control and exclusion) on marsh grass transplants, and factors that can affect them, i.e. the presence or absence of plant neighbors and marsh height (middle and low), revealed that crab herbivory decreased plant biomass and increased stem mortality; however, herbivory effects were significantly diminished in the presence of plant neighbors and with increasing marsh elevation. Our geographical survey showed that crab herbivory is common in SW Atlantic salt marshes, with more than 20 % of leaves damaged in most marshes and with greater consumption at marshes with higher crab densities. In addition, plants at the lower edge of marshes were generally the most consumed (max. >60 % leaves consumed) and crabs preferred S. alterniflora over S. densiflora. Over a regional spatial scale, our results suggest that herbivory may affect plant production at some marshes and can also play a role in limiting the lower tidal elevation limit of low-marsh plants. © Inter-Research 2007.

Descripción: The effects of dopamine (DA) and dopaminergic agonists and antagonists on ion transport were studied in isolated perfused gills of the crab Chasmagnathus granulatus. DA applied under steady state conditions (perfusion with hemolymph-like saline) produced a transient increase of the transepithelial potential difference (Vtc) from 2.2±0.2 to 4.8±0.3 mV, describing an initial cAMP-dependent stimulating phase followed by an inhibitory phase. Spiperone and domperidone (antagonists of D2-like DA receptors in vertebrates) completely blocked the response to DA, while the D1-like antagonist SCH23390 blocked only the inhibitory phase. Theophylline (phosphodiesterase inhibitor) and okadaic acid (protein phosphatases PP1 and PP2A inhibitor) were also able to block the inhibitory phase, suggesting that it depends on adenylyl cyclase inhibition and on protein phosphatases. When the gills were perfused with hypoosmotic solution, or with the adenylyl cyclase activator forskolin, Vte was increased several-fold. DA applied under these stimulated conditions partially reversed the Vte increase by 54% and 25%, respectively. Similarly, the D1-like agonist, fenoldopam, produced a 33% reduction in the stimulated Vte. We propose that, in C. granulatus gills, DA stimulates adenylyl cyclase and therefore ion transport through D1-like receptors linked to a Gs protein, although they respond to antagonists that interact with D2-like receptors in vertebrates. The inhibitory phase seems to be mediated by D2-like receptors linked to a Gi/o protein, which inhibits adenylyl cyclase, although these receptors can be activated or blocked by agonists or antagonists that interact with D1-like receptors in vertebrates and insects.

Descripción: The crab Chasmagnathus granulatus reacts to a shadow passing overhead with an escape response that habituates after 30 trials and for 5 days at least. The effect of a wide range of different intertrial intervals (ITIs) (0, 9, 27, 45, 81, 135, and 171 sec) on the Chasmagnathus long-term habituation (LTH) was evaluated at 24 h. Memory retention was estimated separately at two phases of a six-trial testing session: at first trial (the initial testing phase) and at the subsequent block of five trials (the retraining phase). A training of 30 trials with an ITI equal to or longer than 27 sec induced LTH at both testing phases, however, with a 0- or a 9- sec ITI, training wholly failed to build up LTH. When the number of trials was increased, a massed training (ITI = 0 or 9 sec) induced LTH at re- training but not at initial testing. Thus, massed training produces LTH only at retraining, whereas spaced training (ITI ≤ 27 sec) produces LTH at both initial phase and retraining. An ITI shift from training to testing diminished or abolished retention at retraining regardless of the direction of the shift, thus suggesting that crabs acquire a memory of the trial- spacing at training. According to these results, it is postulated that LTH consists of two memory components: one produced by spaced training and expressed at both initial testing and retraining, and one yielded by massed training and expressed only at retraining. The possibility that the two components of LTH were differentially affected by cycloxemide and context shift is discussed.

Descripción: We studied the transepithelial potential difference (TEPD) and 22Na flux across isolated perfused gills (anterior pair 5 and posterior pairs 6-8) of the crab Chasmagnathus granulatus acclimated to either hypo- or hyper-osmotic conditions. The gills of crabs acclimated to low salinity, perfused and bathed with 10%‰ saline solutions, produced the following TEPDs (hemolymph side with respect to bath side): 0.4±0.7, -10.2±1.6, -10.8±1.3 and -6.7±1.3mV for gills 5, 6, 7 and 8, respectively. Gills 6, 7 and 8 did not differ significantly. Reducing the saline concentration of bath and perfusate from 30%‰ to 20%‰ or 10%‰ increased significantly the TEPDs of these gills. TEPDs of gill 6 (representative of posterior gills) were reduced by 69±5% and 60±5% after perfusion with ouabain or BaCl2 (5mmoll-1 each), respectively. The same gill showed a net ouabain-sensitive Na+ influx of 1150±290 μequiv g-1h-1. Gill 6 of crabs acclimated to high salinity produced TEPDs of -1.5±0.1 and -1.3±0.09mV after perfusion with 30%‰ or 40%‰ salines, respectively. Perfusion with ouabain or BaCl2 reduced TEPDs by 76±7% and 86±4%, respectively. A net ouabain-sensitive Na+ efflux of 2282±337 μequiv g-1h-1 was recorded in gill 6 perfused with 38%‰ saline.

Descripción: Using quantitative real-time PCR, the expression of mRNAs encoding three transport-related proteins and one putative housekeeping protein was analyzed in anterior and posterior gills of the euryhaline crab Chasmagnathus granulatus following transfer from isosmotic conditions (30‰ salinity) to either dilute (2‰) or concentrated (45‰) seawater. Modest changes were observed in the abundance of mRNAs encoding the housekeeping protein arginine kinase and the vacuolar-type H+-ATPase B-subunit, both of which were highly expressed under all conditions. By contrast, the expression of Na +/K+-ATPase α-subunit mRNA and Na+/K +/2Cl- cotransporter mRNA was strongly responsive to external salinity. During acclimation to dilute seawater, cotransporter mRNA increased 10-20-fold in posterior gills within the first 24 h while Na +/K+-ATPase α-subunit mRNA increased 35-55-fold. During acclimation to concentrated seawater, cotransporter mRNA increased 60-fold by 96 h and Na+/K+-ATPase α-subunit increased approximately 25-fold in posterior gills. Our results indicate a complex pattern of transcriptional regulation dependent upon the direction of salinity change and the developmental background of the gills.

Descripción: Experiments with insects and crabs have demonstrated their remarkable capacity to learn and memorize complex visual features (Giurfa et al., 2001; Pedreira and Maldonado, 2003; Chittka and Niven, 2009). Such abilities are thought to require modular brain processing similar to that occurring in vertebrates (Menzel and Giurfa, 2001). Yet, physiological evidence for this type of functioning in the small brains of arthropods is still scarce (Liu et al., 1999, 2006; Menzel and Giurfa, 2001). In the crab Chasmagnathus granulatus, the learning rate as well as the long-term memory of a visual stimulus has been found to be reflected in the performance of identified lobula giant neurons (LGs) (Tomsic et al., 2003). The memory can only be evoked in the training context, indicating that animals store two components of the learned experience, one related to the visual stimulus and one related to the visual context (Tomsic et al., 1998; Hermitte et al., 1999). By performing intracellular recordings in the intact animal, we show that the ability of crabs to generalize the learned stimulus into new space positions and to distinguish it from a similar but unlearned stimulus, two of the main attributes of stimulus memory, is reflected by the performance of the LGs. Conversely, we found that LGs do not support the visual context memory component. Our results provide physiological evidence that the memory traces regarding "what" and "where" are stored separately in the arthropod brain. © 2011 the authors.

Descripción: Background: Human β-amyloid, the main component in the neuritic plaques found in patients with Alzheimer's disease, is generated by cleavage of the β-amyloid precursor protein. Beyond the role in pathology, members of this protein family are synaptic proteins and have been associated with synaptogenesis, neuronal plasticity and memory, both in vertebrates and in invertebrates. Consolidation is necessary to convert a short-term labile memory to a long-term and stable form. During consolidation, gene expression and de novo protein synthesis are regulated in order to produce key proteins for the maintenance of plastic changes produced during the acquisition of new information.Results: Here we partially cloned and sequenced the beta-amyloid precursor protein like gene homologue in the crab Chasmagnathus (cappl), showing a 37% of identity with the fruit fly Drosophila melanogaster homologue and 23% with Homo sapiens but with much higher degree of sequence similarity in certain regions. We observed a wide distribution of cappl mRNA in the nervous system as well as in muscle and gills. The protein localized in all tissues analyzed with the exception of muscle. Immunofluorescence revealed localization of cAPPL in associative and sensory brain areas. We studied gene and protein expression during long-term memory consolidation using a well characterized memory model: the context-signal associative memory in this crab species. mRNA levels varied at different time points during long-term memory consolidation and correlated with cAPPL protein levels. Conclusions: cAPPL mRNA and protein is widely distributed in the central nervous system of the crab and the time course of expression suggests a role of cAPPL during long-term memory formation. © 2010 Fustiñana et al; licensee BioMed Central Ltd.