Morphology and physiology of pleural-to-buccal neurons coordinating defensive retraction with feeding arrest in the pond snail Lymnaea stagnalis

1. We have studied morphology, physiology and chemistry of a bilateral pair of pleural-to-buccal projecting neurons (PlB cells) of the pond snail Lymnaea stagnalis. Intracellular dye fills revealed axon arborization within neuropiles of ipsilateral pedal and cerebral ganglia, as well as in both buccal ganglia. Terminal axons of the left and right PlBs showed close proximity within the buccal commissure. 2. The left and right PlB neurons have been found electrotonically coupled and, sometimes, generating synchronous spikes. 3. The results show that two PlB cells operate as a single unit, and that paired buccal networks responsible for feeding rhythm are treated by the PlBs as a single target.     

Structure of visual pathways in the nervous system of freshwater pulmonate molluscs

Central nervous system of freshwater pulmonate molluscs Lymnaea stagnalis and Planorbarius corneus was stained using retrograde transport of neurobiotin in the optic tract fibers. In both species, perikarya and fibers of the stained neurons are found in all ganglia except the buccal ones. Afferent fibers of the optic nerve form dense sensory neuropil located in relatively small volume of cerebral ganglia. Typical neuronal groups sending their processes into the optic nerves of ipsilateral and contralateral body halves are described. Among them, neurons of visceral and parietal ganglia innervating both eyes concurrently as well as sending projections into peripheral nerves are revealed. These neurons, supposedly, have a function to integrate sensory signals, which may be a basis for regulation of light sensitivity of retina and functioning of peripheral organs. Bilateral links of the molluscan eye with the pedal ganglia cells and statocysts are found, which is, likely, a structural basis of certain known behavioral patterns related to stimulation of visual inputs in the studied gastropod molluscs.     

Multilevel inhibition of feeding by a peptidergic pleural interneuron in the mollusc<Emphasis Type="Italic"> Lymnaea stagnalis</Emphasis>

The pleural interneuron PlB is a white neuron in the pleural ganglion of the snail Lymnaea. We test the hypothesis that it inhibits neurons at all levels of the feeding system, using a combination of anatomy, physiology and pharmacology. There is just one PlB in each pleural ganglion. Its axon traverses the pedal and cerebral ganglia, running into the buccal ganglia. It has neuropilar branches in the regions of the cerebral and buccal ganglia where neurons that are active during feeding also branch. Activation of the PlB blocks fictive feeding, whether the feeding rhythm occurs spontaneously or is driven by a modulatory interneuron. The PlB inhibits all the neurons in the feeding network, including protraction and retraction motoneurons, central pattern generator interneurons, buccal modulatory interneurons (SO, OC), and cerebral modulatory interneurons (CV1, CGC). Only the CV1 interneuron shows discrete 1:1 IPSPs; all other effects are slow, smooth hyperpolarizations. All connections persist in Ca²⁺/Mg²⁺-rich saline, which reduces polysynaptic effects. The inhibitory effects are mimicked by 0.5 to 100 mol l^–1 FMRFamide, which the PlB soma contains. We conclude that the PlB inhibits neurons in the feeding system at all levels, probably acting though the peptide transmitter FMRFamide.Electronic Supplementary Material Supplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s00359-004-0503-x     

Ultrastructure of the Juxtaganglionar Organ,a Putative Endocrine Gland Associated with the Cerebral Ganglia of Aplysia juliana

Summary

Electron microscopy was used to examine the morphology of a putative endocrine gland, the juxtaganglionar organ (JO), and its relation to the cerebral ganglia of the hermaphroditic opisthobranch gastropod Aplysia juliana. The JO is a well-vascularized, poorly innervated tissue of glandular cells—rich in mitochondria, lipids, ribosomes, and endoplasmic reticulum, with sparse cilia and membrane-limited secretory granules—within the connective tissue sheath just exterior to the neuronal soma in the dorsal and posterior portions of the cerebral ganglia. The cytology and organization of the JO supports its homology to the dorsal bodies of pulmonate gastropods, which axe endocrine organs known to release one or more female gonadotropic factors.     

Mercuric chloride-induced Gastrin/Cholecystokinin 8 immunoreactivity in the central nervous system of the terrestrial slug Semperula maculata: an immunohistochemical study

We measured the immunoreactivity of the neuropeptide gastrin cholecystokinin 8 (gastrin/CCK 8) in neurons of the terrestrial slug Semperula maculata following acute treatment with mercuric chloride (HgCl₂). The distribution of gastrin/CCK 8 was analyzed in neurons of different regions, specifically from cerebral ganglia (procerebrum (pro-c), mesocerebrum (meso-c) and metacerebrum (meta-c). In the control group, neurons of pedal, pleural, parietal and visceral ganglia showed positive immunoreactivity using vertebrate antiserum against gastrin/CCK 8. Gastrin/CCK 8 immunoreactivity was also seen in the fibers and neuropil region of all ganglia. In the cerebral ganglion, 10, 12 and 8 % of the neurons from pro-c, meso-c and meta-c, respectively, were stained with the antibody. The immunostaining was increased in neurons (giant, large, medium and small) after HgCl₂ treatment. The treatment greatly increased the mucin content within the neurons. Exposure to HgCl₂ enhanced gastrin immunoreactivity in the neurons and this increased with time. Results are discussed in the context of neuropathology in cerebral ganglia associated with the feeding behavior of Semperula maculata.     

Development of embryonic cells containing serotonin, catecholamines, and FMRFamide-related peptides in Aplysia californica

This study demonstrates the presence of a relatively extensive but previously unrecognized nervous system in embryonic stages of the opisthobranch mollusc Aplysia californica. During the trochophore stage, two pairs of cells were observed to be reactive to antibodies raised against the neuropeptides FMRFamide and EFLRIamide. These cells were located in the posterior region of the embryo, and their anterior projections terminated under the apical tuft. As the embryos developed into veliger stages, serotonin-like immunoreactive (LIR) cells appeared in the apical organ and were later observed to innervate the velum. Also, aldehyde-induced fluorescence indicative of catecholamines was present in cells in the foot, oral, and possibly apical regions during late embryonic veliger stages. Just before the embryo hatches as a free-swimming veliger, additional FMRFamide-LIR and catecholamine-containing cells appeared in regions that correspond to the ganglia of what will become the adult central nervous system (CNS). Neurons and connectives that will contribute to the adult CNS appear to develop along the pathways that are pioneered by the earliest posterior FMRFamide-LIR cells. These observations are consistent with the hypothesis that, besides their presumed roles in the control of embryonic behaviors, some elements may also guide the development of the CNS. Embryonic nervous systems that develop prior to and outside of the adult CNS have also been reported in pulmonate and prosobranch species of molluscs. Therefore, the demonstration of early developing neurons and their transmitter phenotypes in A. californica presents new opportunities for a better understanding of the ontogeny and phylogeny of both behavioral and neuronal function in this important model species.     

Characterization of cardiac innervation in the nudibranch,Archidoris montereyensis

Summary The heart of the nudibranch mollusc Archidoris montereyensis is regulated by a small number of powerful effector neurons located in the right pleural and visceral ganglia. Two identifiable neurons in the pleural ganglion, a heart excitor (pl_HE) and a heart inhibitor (Pl_HI), are especially important regulators of cardiac function in that low levels of spontaneous activity in either cell significantly alters the amplitude and rate of heart contractions. These neurons have extensive dendritic arbors within the right pleural ganglion and branching axonal processes within the visceral ganglion. The visceral ganglion also contains a heart excitor neuron (V_HE) and at least two heart inhibitor neurons (V_HI cells), but their influence on cardiac activity is weaker than that of the pleural ganglion cells. All of these heart effector cells appear to be motor neurons with axons that terminate predominately in the atrio-ventricular valve region of the heart via the pericardial nerve. The simplicity and strength of these neuronal connections to the heart of Archidoris make this a favorable preparation for studies of cardiac regulation.Abbreviations Pl _HE pleural ganglion heart excitor neuron - Pl _HI pleural heart inhibitor neuron - V _HE visceral ganglion heart excitor neuron - V _HI cells, visceral heart inhibitor neurons - V _K visceral kidney excitor neuron - V _G visceral gill excitor neuron     

Nitrite and Nitrate Levels in Individual Molluscan Neurons: Single-Cell Capillary Electrophoresis Analysis

Abstract: Cell and tissue concentrations of NO₂^? and NO₃^? are important indicators of nitric oxide synthase activity and crucial in the regulation of many metabolic functions, as well as in nonenzymatic nitric oxide release. We adapted the capillary electrophoresis technique to quantify NO₂^? and NO₃^? levels in single identified buccal neurons and ganglia in the opisthobranch mollusc Pleurobranchaea californica, a model system for the study of the chemistry of neuron function. Neurons were injected into a 75-µm separation capillary and the NO₂^? and NO₃^? were separated electrophoretically from other anions and detected by direct ultraviolet absorbance. The limits of detection for NO₂^? and NO₃^? were <200 fmol (<4 µM in the neurons under study). The NO₂^? and NO₃^? levels in individual neurons varied from 2 mM (NO₂^?) and 12 mM (NO₃^?) in neurons histochemically positive for NADPH-diaphorase activity down to undetectable levels in many NADPH-diaphorase-negative cells. These results affirm the correspondence of histochemical NADPH-diaphorase activity and nitric oxide synthase in molluscan neurons. NO₂^? was not detected in whole ganglion homogenates or in hemolymph, whereas hemolymph NO₃^? averaged 1.8 ± 0.2 × 10^?3M. Hemolymph NO₃^? in Pleurobranchaea was appreciably higher than values measured for the freshwater pulmonate Lymnaea stagnalis (3.2 ± 0.2 × 10^?5M) and for another opisthobranch, Aplysia californica (3.6 ± 0.7 × 10^?4M). Capillary electrophoresis methods provide utility and convenience for monitoring NO₂^?/NO₃^? levels in single cells and small amounts of tissue.     

Light- and electron-microscopic immunocytochemistry of a molluscan insulin-related peptide in the central nervous system of Planorbarius corneus

Summary Two groups of cerebral dorsal cells of the pulmonate snail Planorbarius corneus stain positively with antisera raised against synthetic fragments of the B- and C-chain of the molluscan pro-insulin-related prohormone, proMIP-I, of another pulmonate snail, Lymnaea stagnalis. At the light-microscopic level the somata of the dorsal cells and their axons and neurohemal axon terminals in the periphery of the paired median lip nerves are immunoreactive with both antisera. Furthermore, the canopy cells in the lateral lobes of the cerebral ganglia are positive. In addition, MIP_B-immunoreactive neurons are found in most other ganglia of the central nervous system. At the ultrastructural level, pale and dark secretory granules are found in somata and axon terminals of the dorsal cells. Dark granules are about 4 times as immunoreactive to both antisera as pale granules. Release of anti-MIP_B- and anti-MIP_C-immunopositive contents of the secretory granules by exocytosis is apparent in material treated according to the tannic acid method. It is concluded that the dorsal and canopy cells synthesize a molluscan insulin-related peptide that is packed in the cell body into secretory granules and that is subsequently transported to the neurohemal axon terminals and released into the hemolymph by exocytosis. Thus, MIP seems to act as a neurohormone on peripheral targets. On the basis of the analogy between the dorsal cells and the MIP-producing cells in L. stagnalis, it is proposed that the dorsal cells of P. corneus are involved in the control of body growth and associated processes.     

后鳃亚纲软体动物化学防御物质研究进展

海洋后鳃亚纲软体动物（Opisthobranchia）属软体动物门（Mollusca）复足纲软体动物（Gastropoda）,其成体体表的被壳或退化或完全消失。由于失去了物理保护,海洋后鳃亚纲软体动物的生存主要依赖于化学防御机制。大多数海洋后鳃亚纲软体动物通过选择适当的食物,并将其中有用的代谢物质经过进一步生物转化或积累到身体的特定部位作为化学防御性物质,以保护自己不受天敌的捕食;少数动物能够生物合成自身所需要的化学物质,从而建立其化学防御体系。显然,研究后鳃亚纲软体动物及其食源生物的化学组成,可以揭示它们之间的食物链关系,并进一步阐明这些化学物质的生态学作用。由于相同软体动物在食性上具有统一性,因此对其体内化学成分的分析也有助于其分类学的研究。同时,这种进化的化学防御体系为我们提供了一条从自然界寻找生物活性物质的新方法。基于以上原因,来自生态学、化学以及药理学等不同领域的科学家均对海洋后鳃亚纲软体动物表现出极大的兴趣,并对其进行了一系列研究;在过去的20多年中,发表了大量的相关论文。综述了海洋后鳃亚纲软体动物近5a来的研究概况,文章涵盖了所有研究的3个大的种群,旨在向读者介绍有关该类动物的化学研究情况,并据此讨论其可能的生态作用。     

Electrophysiological study of serotoninergic neuron Cl in the pteropod mollusk Clione

Functional characteristics of cerebral serotoninergic neuron Cl, axons of which terminate at the buccal ganglia [7], were investigated in the pteropod molluskClione. Stimulating neuron Cl induced activation of the feeding rhythm generator located in the buccal ganglia — an effect arising after a long latency and persisting for some tens of seconds once stimulation had ended. Neuron Cl receives feedback from buccal ganglion cells and this brings about periodic modulation in ganglia activity during the generation of feeding rhythm. Activity of neuron Cl is correlated with operation of the locomotor rhythm generator located in the pedal ganglia. The firing rate of Cl neurons increased upon activation of the locomotor generator (whether spontaneous or induced by stimulating certain command neurons). The correlation found between workings of the locomotor generator and activity of Cl neurons is thought to be one of the manifestations of feeding synergy involving simultaneous activation of the locomotor and buccal apparatus.Institute for Research on Information Transmission, Academy of Sciences of the USSR, Moscow. M. V. Lomonosov State University, Moscow. Translated from Neirofiziologiya, Vol. 23, No. 1, pp. 18–25, January–February, 1991.     

The feeding habits of Sargochromis Codringtonii in Lake Kariba, Zimbabwe

The food of adult Sargochromis codringtonii in Lake Kariba was dominated by prosobranch snails with pulmonate snails forming an insignificant dietary component. S. codringtonii does not show particular preference for any snail species, feeding on those species that are readily available. Morphometrical analysis of the feeding structures of S. codringtonii showed that the fish is well adapted to handling a molluscivorous diet. The pharyngeal jaws with their flat crowned teeth are well adapted to crushing molluscs. The daily food consumption rate of S. codringtonii was estimated over different seasons. Estimates of food consumption indicate that the fish consumed 13.96% of their dry body weight per day in summer falling to 4.46% in winter. The low consumption of pulmonate snails suggests that S. codringtonii can not be a successful biological control agent against the vectors of schistosomiasis.     

Neurons determining passive defensive response in the pteropod mollusk Clione limacina

Neurons responding to tactile stimulation of the head with bursts of action potentials of short latency followed by passive defensive response were found in the pedal ganglia and identified as Pd13. Stimulation of one Pd13 neuron leads to inhibition of the entire locomotor generator. A whole set of neurons, identified as P2, 3, 4, and 5, activated solely by intensive tactile stimulation of the head, were found in the pleural ganglia. Stimulating one such neuron also induces inhibition of the entire locomotor generator. These pleural cells are synaptically connected with Pd13 neurons and one EPSP in Pd13 unit corresponds to each action potential in the pleural cell. This connection has a facility for potentiation, subsequently replaced by habituation. In this way, pleural neurons also introduce Pd13 neurons into the inhibitory trend when activated by intensive tactile stimulation. Application of cerucal and ergotamine (dopaminergic receptor blockers) suppresses the inhibitory effect of the Pd13 neuron and pleural cells, thus indicating dopamine involvement in the inhibitory processes occurring in passive defensive reaction.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 21, No. 5, pp. 685–694, September–October, 1989.     

Prey capture phase of feeding behavior in the pteropod mollusc,clione limacina: neuronal mechanisms

The prey capture phase of feeding behavior in the pteropod mollusc Clione limacina consists of an explosive extrusion of buccal cones, specialized structures which are used to catch the prey, and acceleration of swimming with frequent turning and looping produced by tail bend. A system of neurons which control different components of prey capture behavior in Clione has been identified in the cerebral ganglia. Cerebral B and L neurons produce retraction of buccal cones and tightening of the lips over them — their spontaneous spike activities maintain buccal cones in the withdrawn position. Cerebral A neurons inhibit B and L cells and produce opening of the lips and extrusion of buccal cones. A pair of cerebral interneurons C-BM activates cerebral A neurons and synchronously initiates the feeding motor program in the buccal ganglia. Cerebral T neurons initiate acceleration of swimming and produce tail bending which underlies turning and looping during the prey capture. Both tactile and chemical inputs from the prey produce activation of cerebral A and T neurons. This reaction appears to be specific, since objects other than alive Limacina or Limacina juice do not initiate activities of A and T neurons.     

Multi‐directional sensitivity of statocyst receptor cells of the opisthobranch gastropod Aplysia limacina

The electrical activity in the static nerves of Aplysia limacina was studied in relation to the spatial orientation of the animal. Each sense cell shows a multi‐directional sensitivity. Hence, it responds during full‐circle rotations about all horizontal axes but only within a limited angular range. The statocyst receptor cells are found to be gravity receptors only, and their electrical behaviour suggests that main information about the spatial orientation of Aplysia is contained in the pattern of responding cells, i.e. which of the 13 cells are active at any one time, and not in their absolute discharge frequencies. Furthermore, evidence is given to a similar function of the statocysts of opisthobranch and pulmonate gastropods.     

Evidence for homologous peptidergic neurons in the buccal ganglia of diverse nudibranch mollusks

The buccal ganglia of seven nudibranches (Aeolidia papillosa, Armina californica, Dirona albolineata, D. picta, Hermissenda crassicornis, Melibe leonina, and Tritonia diomedea) were examined to explore possible homologies between large cells that reacted with antibodies directed against small cardioactive peptide B (SCP_B). The buccal ganglion of each species possessed a pair of large, dorsal–lateral, whitish neurons that contained an SCP_B-like peptide. We refer to these neurons as the SLB (SCP_B-immunoreactive Large Buccal) cells. In all species examined, the SLB cells project out the gastroesophageal nerves and appear to innervate the esophagus. In each species, an apparent rhythmic feeding motor program (FMP) was observed by intracellular recording from both SLB neurons and other neurons in isolated preparations of the buccal ganglia. SLB cells often fire at a high frequency, and usually burst in a specific phase relation to the FMP activity. Stimulation of SLB cells enhances expression of the feeding motor program, either by potentiating existing activity or eliciting the FMP in quiescent preparations. Finally, perfusion of isolated buccal ganglia with SCP_B excites the SLB cells and activates FMPs. Thus, both the immunohistochemical and electrophysiological data suggest that the SLB cells within three suborders of the opistobranchia (Dendronotacea, Arminacea, and Aeolidacea) are homologous. A comparison of our data with previously published studies indicates that SLB cell homologs may exist in other gastropods as well.     

Comparative pharmacology of feeding in molluscs

1. This paper reviews the role of transmitters in identified neurons of gastropod molluscs in generating and modulating fictive feeding. 2. In Lymnaea and Helisoma the 3 phase rhythm is generated by sets of interneurons which use acetylcholine for the N1 (protraction) phase, glutamate for the N2 (rasp) phase interneurons. The N3 interneurons are likely to use several different transmitters, of which one is octopamine. 3. In all the species examined, serotonin (5-HT) is released from giant cerebral cells. Other amines, including dopamine and octopamine, are present in the buccal ganglia and all these amines activate or enhance feeding. 4. Nitric oxide (NO), mostly originating from sensory processes, can also activate fictive feeding, but (at least in Lymnaea) may also be released centrally from buccal (B2) and cerebral neurons (CGC). 5. The central pattern generator for feeding is also modulated by peptides including APGWamide, SCP(B) and FMRFamide. 6. There is increasing evidence that most of these transmitters/modulators act on feeding neurons through second messenger systems--allowing them to act as longer-lasting neuromodulators of the feeding network. 7. Many of the transmitters are used in similar ways by each of the gastropods examined so far, so that their function in the CNS seems to have been conserved through evolution.