Detection of substrate-associated odour cues versus prey-associated cues by the oral veil in Tritonia diomedea

Our goal was to test two potential sensory roles for the oral veil in the nudibranch Tritonia diomedea (now synonymous with T. tetraquetra). First, we hypothesized this cephalic sensory organ could detect substrate-associated odours left behind by an odour plume flowing across sediment. In two experiments in a laboratory flow tank, however, T. diomedea did not show consistent crawling headings in response to either prey or predator odours associated with sediment substrate. In one of the experiments, the slugs did significantly decrease crawling speed in response to prey odours. Although slugs could thus detect at least some substrate-associated odours, these results suggest such cues are not used for navigation. We next considered the oral veil’s potential role in behaviours requiring responses to nearby cues. Our observations of animals before and after denervation of the oral veil suggest that, unsurprisingly, predatory bite-strikes do rely on sensory input from the oral veil. Overall, these data, combined with the results of earlier studies, are consistent with the oral veil detecting cues primarily from nearby stimuli (including both chemical and mechanical modalities), while having little or no role in detecting and responding to odour cues originating from distant sources used for navigation behaviour.     

Serotonin inhibits ciliary transport in esophagus of the nudibranch mollusk Tritonia diomedea

Both serotonin and the molluskan pedal neuropeptides (TPEPs) cause increased ciliary beating rate of cells of the foot epithelium of the nudibranch mollusk, Tritonia diomedea. Here we compared responses of the ciliated epithelium of the esophagus with that of the foot, and report fundamental differences. Serotonin reduces the ciliary transport rate of the esophagus. We find also that the serotonin driven inhibition of esophagus is blocked and the excitation of foot epithelium is reduced by the serotonin receptor blocker ketanserin. On the contrary, ergometrine completely blocked the serotonin effect in the esophagus, and does not block the serotonin effect in the foot. Neither the TPEP driven excitation of ciliated cells of the foot nor that of the esophagus is blocked by ketanserin and ergometrine. Clearly, serotonin and TPEP regulation of different ciliated epithelia involve different receptors. Thus, mechanisms of serotonin control of different ciliated epithelia in the same animal are apparently fundamentally different, and unlike responses in all previous reports, 5HT here inhibits a ciliated epihelium.     

Orientation and navigation relative to water flow, prey, conspecifics, and predators by the nudibranch mollusc Tritonia diomedea

Progress in understanding sensory and locomotory systems in Tritonia diomedea has created the potential for the neuroethological study of animal navigation in this species. Our goal is to describe the navigational behaviors to guide further work on how the nervous system integrates information from multiple senses to produce oriented locomotion. Observation of T. diomedea in its habitat has suggested that it uses water flow to navigate relative to prey, predators, and conspecifics. We test these hypotheses in the field by comparing slug orientation in time-lapse videos to flow direction in circumstances with and without prey, predators, or conspecifics upstream. T. diomedea oriented upstream both while crawling and after turning. This trend was strongest before feeding or mating; after feeding or mating, the slugs did not orient significantly to flow. Slugs turned downstream away from an upstream predator but did not react in control situations without an upstream predator. These data support the hypothesis that T. diomedea uses a combination of odors (or some other cue transported downstream) and water flow to navigate relative to prey, predators, and conspecifics. Understanding the context-dependent choice between upstream and downstream crawling in T. diomedea provides an opportunity for further work on the sensory integration underlying navigation behavior.     

Immunochemical and electrophysiological analyses of magnetically responsive neurons in the mollusc Tritonia diomedea

Tritonia diomedea uses the Earth’s magnetic field as an orientation cue, but little is known about the neural mechanisms that underlie magnetic orientation behavior in this or other animals. Six large, individually identifiable neurons in the brain of Tritonia (left and right Pd5, Pd6, Pd7) are known to respond with altered electrical activity to changes in earth-strength magnetic fields. In this study we used immunochemical, electrophysiological, and neuroanatomical techniques to investigate the function of the Pd5 neurons, the largest magnetically responsive cells. Immunocytochemical studies localized TPeps, neuropeptides isolated from Pd5, to dense-cored vesicles within the Pd5 somata and within neurites adjacent to ciliated foot epithelial cells. Anatomical analyses revealed that neurites from Pd5 are located within nerves innervating the ipsilateral foot and body wall. These results imply that Pd5 project to the foot and regulate ciliary beating through paracrine release. Electrophysiological recordings indicated that, although both LPd5 and RPd5 responded to the same magnetic stimuli, the pattern of spiking in the two cells differed. Given that TPeps increase ciliary beating and Tritonia locomotes using pedal cilia, our results are consistent with the hypothesis that Pd5 neurons control or modulate the ciliary activity involved in crawling during orientation behavior.     

Sperm tail differentiation in the nudibranch mollusc Hypselodoris tricolor (gastropoda,opisthobranchia)

The sperm axoneme of Hypselodoris tricolor forms from a single centriole that is located initially beneath the plasma membrane and then migrates to the nuclear surface. A conspicuous centriolar adjunct-like formation is present in the neck of midspermatids, but it becomes very reduced at the end of spermiogenesis. In spermatocyte and spermatid mitochondria, intracristal bodies originate from the accumulation of a dense material in some cristae. From our observations and foregoing reports, it may be concluded that the process of sperm tail differentiation in opisthobranchs resembles that in pulmonates, whereas it differs in many respects from that occurring in prosobranchs. The appearance of intracristal bodies in modified mitochondria seems to be a special feature of spermatogenesis in the opisthobranchs that does not occur in the two other groups of gastropod molluscs.     

Pharmacological manipulation of serotonin levels in the nervous system of the opisthobranch mollusc Tritonia diomedea

Serotonin-related disorders can be treated by manipulating serotonin synthesis with the serotonin precursor 5-hydroxytryptophan (5-HTP) or other pharmacological agents. The mollusc Tritonia diomedea is a model for investigating the effects of altering serotonin content on the functions of identified neurons. We used high-performance liquid chromatography and immunohistochemistry to examine the amount and localization of 5-HTP, serotonin, and the serotonin breakdown product 5-hydroxyindolacetic acid (5-HIAA) in the Tritonia brain after various pharmacological treatments. Exposure to 5-HTP (2 mM for 30 min-1 h) caused an immediate and massive increase in total 5-HTP content, which lasted more than 20 h, and the widespread appearance of 5-HTP immunoreactivity in neurons. Serotonin levels rose gradually, but only a restricted number of additional neurons displayed serotonin immunoreactivity. 5-HTP treatment also caused an increase in the total amount of 5-HIAA and the appearance of 5-HIAA immunoreactivity throughout the brain. Treatment with the synthesis cofactor tetrahydrobiopterin, the initial precursor tryptophan, or serotonin itself had no persistent effect on total serotonin content. The amino acid decarboxylase inhibitor hydroxybenzylhydrazine (NSD-1015) also had no effect on the total serotonin content, although it caused an accumulation of 5-HTP. Thus, serotonin levels in the brain of T. diomedea appear to be maintained by a homeostatic mechanism that can be disrupted by 5-HTP.     

Function of identified nerves in orientation to water flow in Tritonia diomedea

We determined which sensory and motor nerves mediate orientation to flow in the marine slug Tritonia diomedea, and tested the hypothesis that the slug orients to water flow by comparing the intensities of water flow stimulation on each side of its body. Lesion experiments revealed which nerves carried information necessary for flow orientation. The lateral branches of Cerebral Nerve # 2 were the only cerebral nerves necessary for flow orientation. Cutting all cerebral nerves except the lateral branches of Cerebral Nerve # 2 did not eliminate flow orientation. Thus, the lateral branches of Cerebral Nerve # 2 were both necessary and sufficient (among the cerebral nerves) for flow orientation. Denervation of one side of the head by cutting Cerebral Nerves # 1–4 on one side did not eliminate normal flow orientation. We have revised our model of how Tritonia diomedea orients to flow to allow for this unilateral determination of flow direction. Unilaterally cutting Pedal Nerve # 3, which contains many pedal motor axons, reduced turning toward that side, but did not affect final orientation to flow. The ability to detect flow direction was not compro mised by the inability to initially turn towards flow.Abbreviations CeN cerebral nerve - PeN pedal nerve - PlN pleural nerve     

Morphology of two pairs of identified peptidergic neurons in the buccal ganglia of the mollusc Tritonia diomedea

The morphology of two pairs of identified peptidergic neurons (B11 and B12) located in the buccal ganglia of Tritonia diomedea was described. Both pairs of neurons contained a large quantity of a small cardioactive peptide (SCP) in their somata. One of the pairs (B11), the large dorsal white cells, contained ACh in their somata along with SCP. Both pairs of cells appeared white in live preparations under epi-illumination. Each B11 and B12 was a unipolar neuron and sent its major axonal branch through the ipsilateral gastro-esophageal nerve to the gut. In addition, B12 sent a small branch to the contralateral buccal ganglion. A characteristic feature of both neuron pairs was their vesicular content. Three types of vesicles were observed in B11. Vesicles with electron-lucent core (LCV) and electron-dense core (DCV) were found in the somata. The axon hillock and the beginning of axon contained vesicles with variable electron dense core (VDCV) in addition to LCV and DCV. The ratio of DCV: LCV: VDCV changed from 5:95:0 for the perinuclear cytoplasm to 8:55:37 for the beginning of axon. The average maximum diameters were 97 +/- 23 nm for DCV, 103 +/- 32 nm for LCV and 106 +/- 29 nm for VDCV. B12 somata contained DCV (average maximum diameter 100 +/- 26 nm), LCV (109 +/- 23 nm) and elliptical vesicles with eccentric electron-opaque core (115 +/- 20 nm).     

Nuclear morphogenesis during spermiogenesis in the nudibranch molluscHypselodoris tricolor (Gastropoda,opisthobranchia)

An electron microscope study was carried out on Hypselodoris tricolor spermatids to describe the development of the nuclear morphogenesis and investigate the possible cause(s) of the change in the shape of the spermatid nucleus during spermiogenesis. Three different stages may be distinguished in the course of the nuclear morphogenesis on the basis of the morphology and inner organization of the nucleus. Stage 1 spermatid nuclei are spherical or ovoid in shape and the nucleoplasm finely granular in appearance. Stage 2 nuclei exhibit a disc- or cup-shaped morphology, and the chromatin forms short, thin filaments. During stage 3, a progressive nuclear elongation takes place, accompanied by chromatin rearrangement, first into fibers and then into lamellae, both formations helically oriented. A row of microtubules attached to the nuclear envelope completely surrounds the nucleus. Interestingly, the microtubules always lie parallel to the chromatin fibers adjacent to them. Late stage 3 spermatids show the highest degree of chromatin condensation and lack the manchette at the end of spermiogenesis. Our findings indicate the existence of a clear influence exerted on the chromatin by the manchette microtubules, which appear to be involved in determining the specific pattern of chromatin condensation in Hypselodoris tricolor.     

Longitudinal organisation of the mollusc species in a theoretical French river

An analysis of mollusc communities collected at 272 stations was carried out, using multivariate statistical methods. The stations ranged from the sources to the lower reaches of large rivers. Nine malacotypes succeeded each other along this upstream-downstream gradient; comments are given regarding the composition of their communities. The organisation of the 52 mollusc species included corresponds to that of a theoretical river, representative of the French hydrographic system. The reduction of the abundance and of species richness in the lower reaches of large rivers is discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Neuropeptide TPep action on salivary duct ciliary beating rate in the nudibranch molluscTritonia diomedea

Recently, in the marine molluscTritonia, a family of three peptides (TPep-NLS,-PLS,-PAR) from identified pedal ganglion neurons has been characterized and shown to regulate ciliary beat frequency in epithelia and isolated cells of the molluscan foot. In this study, using an antiserum raised against TPep-NLS, immunofluorescent labelling was observed in specific nerve cell bodies and axons in the buccal ganglia ofTritonia, as well as in axons leading to and innervating the salivary ducts, salivary glands, oesophagus and foregut. This pattern of innervation suggests that buccal ganglion neurons containing TPep control the beating rate of ciliated cells in feeding organs. Accordingly, TPeps were introduced to isolated ciliated salivary ducts. It was found that TPeps and serotonin increased the ciliary beat frequency of cells of the salivary duct similarly; other peptides (such as APep fromAplysia) had no such effect. Threshold sensitivity both for TPeps and serotonin was approximately 10⁻⁸ M, with maximal response occurring above 10⁻⁵ M. Taken together, these structural and physiological results suggest that TPep-like peptides are present in the salivary and other feeding organs ofTritonia and are involved in the regulation of salivary transport.     

Surface charges on the outer side of mollusc neuron membrane

Summary The shifts of current-voltage characteristics of sodium and calcium inward currents produced by changes in the concentration of divalent cations (Mg²⁺, Ca²⁺, Sr²⁺, Ba²⁺) and in pH of the extracellular solution have been measured on isolated neurons of the molluscHelix pomatia intracellularly perfused with potassium-free solutions. On the basis of these shifts and using Stern's theory (O. Stern, 1924.Z. Electrochem. 30508–516), the binding constants for the ions to charged groups of the outer side of the somatic membrane and the density of the surface charges produced by these groups have been calculated. For groups located in the vicinity of sodium channels we obtainedK _Ca=90±10,K _Sr=60±10,K _Ba=25±5 andK _Mg=16±5m ^–1 at pH=7.7 and for groups located in the vicinity of calcium channelsK _Ca=67±10,K _Sr=20±5 andK _Ba=19±5m ^–1 at pH=7.0. The same groups bind H⁺ ions with apparent pK=6.2±0.2 that corresponds toK _H=1.6×10⁶ m ^–1. The density of fixed charges near the sodium channels is 0.17±0.05 e/nm² (pH=7.7) and near the calcium channels is 0.23±0.05 electrons/nm² (pH=7.0). From the comparison of the obtained values with the data about binding constants of the same ions to different negatively charged phospholipids, a suggestion is made that just the phophatidylserine is responsible for the surface potential of the outer side of the somatic membrane. It was also shown that the presence of this potential results in a change in the concentration of carrier ions near the membrane which affects the maximal values of the corresponding transmembrane currents.     

Nucleosomal organization of chromatin in sperm nuclei of the bivalve mollusc Aulacomya ater

The sperm nuclei of Aulacomya ater, family Mitylidae, contain three proteins (X, Aa5 and Aa6) which are specific to this cell type coexisting with a set of five somatic-type histones. Information about the chromatin structure resulting from this kind of association is scarce. Therefore, we have probed the structure of this sperm chromatin through digestion with micrococcal nuclease in combination with salt fractionation. The data obtained have allowed us to propose a nucleosomal arrangement for this chromatin. However, two types of nucleosomes would be present in agreement with their protein components.     

大亚湾潮间带软体运动的物种多样性初步研究

1999年7月在大亚湾潮间带4种不同生境（岩石岸、砾石滩、泥沙滩、沙滩）10个样方采集软体动物38种,隶属于23个科。采用丰富度指数和多样性指数对不同生境软体动物的多样性进行研究。研究结果表明,大亚湾软体动物物种丰富度指数砾石滩、岩石岸(D=4.328～9.378)>泥沙滩(D=4.328～6.493)>沙滩(D=2.886)。多样性指数也显示砾石滩、岩石岸(H′=0.429～0.842)>泥沙滩(H′=0.315～0.450)>沙滩(H′=0.182)。对上述样方聚类结果表明,软体动物种类分布及数量受底质、浪击和污染的影响。     

Morphology of wing mechanoreceptors involved in the wing retraction reflex in the pteropod mollusc Clione limacina

Tactile stimulation of the wings (parapodia) of actively swimming Clione limacina results in inhibition of swimming and retraction of the wings. Electrophysiological evidence suggests that wing mechanoreceptors have central cell bodies and wide innervation fields in the ipsilateral wing. Scanning electron microscopy of expanded wings reveals ciliary cone processes arranged in a pattern that is similar to the electrophysiologically‐determined innervation fields of wing mechanoreceptors. Transmission electron microscopy suggests that the ciliary cone structures are terminal processes of neuron‐like cells. Three‐dimensional reconstructions of serially‐sectioned terminal processes indicate that cell bodies are not found in the wing epithelium or immediately under the epithelium, further supporting the notion that the wing mechanoreceptors have central cell bodies.