Serotonin catabolism depends upon location of release: characterization of sulfated and gamma-glutamylated serotonin metabolites in Aplysia californica

Serotonin is a vital neurotransmitter for the functioning of the nervous system in species throughout the animal phyla. Despite its ubiquitous nature, the metabolism of this molecule has yet to be completely elucidated in even the most basic of organisms. Two novel serotonin catabolites, serotonin-O-sulfate and gamma-glu-serotonin-O-sulfate, are chemically characterized using capillary electrophoresis with wavelength-resolved fluorescence detection and electrospray mass spectrometry, and the formation of gamma-glu-serotonin in Aplysia californica is confirmed. These novel compounds appear to be synthesized enzymatically, and known mammalian enzymes exist for all serotonin transformations observed here. The pathway of serotonin inactivation depends upon the type of neuronal tissue subjected to neurotransmitter incubation, with assorted serotonin products observed in distinct locations. Initially demonstrated to be in the metacerebral cell (MCC) soma, the new serotonin metabolite serotonin-O-sulfate may contribute to important functions in the serotonergic system beyond simple serotonin inactivation.     

Copper induced oxidation of serotonin: analysis of products and toxicity

Serotonin is a major neurotransmitter that controls many functions, ranging from mood and behaviour through to sleep and motor functions. The non-enzymatic oxidation of serotonin is of significant importance as some oxidation products are considered to be neurotoxic. An interaction between copper and serotonin has been suggested by symptoms observed in a number of neurodegenerative diseases such as Wilson's and Prion diseases. Using PC12 cells as a model of neuronal cells, we show that the interaction between copper and serotonin is toxic to undifferentiated cells. The toxicity is largely due to reactive oxygen species as cell death is significantly reduced in the presence of the antioxidant mannitol. Differentiation of the PC12 cells also confers resistance to the oxidative process. In vitro oxidation of serotonin by copper results in the eventual formation of a coloured pigment, thought to be a melanin-like polymeric species. Using spectroscopic methods we provide evidence for the formation of a single intermediate product. This dimeric intermediate was identified and characterized as 5,5'-dihydroxy-4,4'-bitryptamine. These results indicate that copper structurally alters serotonin and this process may play a role in copper related neurodegenerative diseases.     

A major clade of prokaryotes with ancient adaptations to life on land

Evolutionary trees of prokaryotes usually define the known classes and phyla but less often agree on the relationships among those groups. This has been attributed to the effects of horizontal gene transfer, biases in sequence change, and large evolutionary distances. Furthermore, higher level clades of prokaryote phyla rarely are supported by information from ecology and cell biology. Nonetheless, common patterns are beginning to emerge as larger numbers of species are analyzed with sophisticated methods. Here, we show how combined evidence from phylogenetic, cytological, and environmental data support the existence of an evolutionary group that appears to have had a common ancestor on land early in Earth's history and includes two-thirds of known prokaryote species. Members of this terrestrial clade (Terrabacteria), which includes Cyanobacteria, the gram-positive phyla (Actinobacteria and Firmicutes), and two phyla with cell walls that differ structurally from typical gram-positive and gram-negative phyla (Chloroflexi and Deinococcus-Thermus), possess important adaptations such as resistance to environmental hazards (e.g., desiccation, ultraviolet radiation, and high salinity) and oxygenic photosynthesis. Moreover, the unique properties of the cell wall in gram-positive taxa, which likely evolved in response to terrestrial conditions, have contributed toward pathogenicity in many species. These results now leave open the possibility that terrestrial adaptations may have played a larger role in prokaryote evolution than currently understood.     

Reduced female mating receptivity and activation of oviposition in two Callosobruchus species due to injection of biogenic amines

Analyses of proximate mechanisms that control mating and oviposition behaviours in insects are important because they link behavioural ecology and physiology. Recently, seed beetles have been used as models to study evolution of female multiple mating and cost of reproduction including mating. In the present study, we investigated the effects of biogenic amines into the abdomens of females of two Callosobruchus species, Callosobruchus chinensis and Callosobruchus maculatus, on mating receptivity and oviposition behaviour. In C. chinensis, injection of octopamine and tyramine reduced receptivity to mating and tyramine and serotonin increased the number of eggs laid. Similarly, injection of tyramine reduced the receptivity of females and increased the number of eggs laid by females of C. maculatus. These results show the possibility that biogenic amines control mating receptivity and oviposition behaviour in females of two Callosobruchus species.     

Different functions for homologous serotonergic interneurons and serotonin in species-specific rhythmic behaviours

Closely related species can exhibit different behaviours despite homologous neural substrates. The nudibranch molluscs Tritonia diomedea and Melibe leonina swim differently, yet their nervous systems contain homologous serotonergic neurons. In Tritonia, the dorsal swim interneurons (DSIs) are members of the swim central pattern generator (CPG) and their neurotransmitter serotonin is both necessary and sufficient to elicit a swim motor pattern. Here it is shown that the DSI homologues in Melibe, the cerebral serotonergic posterior-A neurons (CeSP-As), are extrinsic to the swim CPG, and that neither the CeSP-As nor their neurotransmitter serotonin is necessary for swim motor pattern initiation, which occurred when the CeSP-As were inactive. Furthermore, the serotonin antagonist methysergide blocked the effects of both the serotonin and CeSP-As but did not prevent the production of a swim motor pattern. However, the CeSP-As and serotonin could influence the Melibe swim circuit; depolarization of a cerebral serotonergic posterior-A was sufficient to initiate a swim motor pattern and hyperpolarization of a CeSP-A temporarily halted an ongoing swim motor pattern. Serotonin itself was sufficient to initiate a swim motor pattern or make an ongoing swim motor pattern more regular. Thus, evolution of species-specific behaviour involved alterations in the functions of identified homologous neurons and their neurotransmitter.     

Current problems in marine biodiversity studies

This paper deals with a discussion of terminology and six proposed levels of biodiversity. Recent data and estimates were used to compare species and taxonomic diversity of terrestrial, freshwater, and marine organisms. About 1.5 million terrestrial species and 320000 aquatic species are hitherto known. In spite of a long history of research, only about 280000 marine species have been discovered, of which 180000 species are invertebrates. Of 33 metazoan phyla, 31 are found in the sea, 13 of these being exclusively marine. Seventeen metazoan phyla contain freshwater species, and only 11 phyla comprise terrestrial animals. Two phyla (freshwater Micrognathozoa and terrestrial Onychophora) possess no marine species. In this paper, we review the assessment reports on marine biological diversity in coral reefs, coastal ecosystems, macrobenthos, and meiofauna. Recent data on the number of known species are listed for each metazoan phylum; the number of anticipated new species to be discovered is estimated. Deep-sea macrobenthos are believed to comprise about 25 million species; meiofauna seems to be composed of 20 to 30 million species, ten million of whom are marine nematodes. Hypotheses are discussed that can account for the high species diversity of deep-sea macrobenthos and meiofauna.     

A review of FMRFamide- and RFamide-like peptides in metazoa

Neuropeptides are a diverse class of signalling molecules that are widely employed as neurotransmitters and neuromodulators in animals, both invertebrate and vertebrate. However, despite their fundamental importance to animal physiology and behaviour, they are much less well understood than the small molecule neurotransmitters. The neuropeptides are classified into families according to similarities in their peptide sequence; and on this basis, the FMRFamide and RFamide-like peptides, first discovered in molluscs, are an example of a family that is conserved throughout the animal phyla. In this review, the literature on these neuropeptides has been consolidated with a particular emphasis on allowing a comparison between data sets in phyla as diverse as coelenterates and mammals. The intention is that this focus on the structure and functional aspects of FMRFamide and RFamide-like neuropeptides will inform understanding of conserved principles and distinct properties of signalling across the animal phyla.     

Host sharing and host manipulation by larval helminths in shore crabs: cooperation or conflict?

Larval helminths of different species that share the same intermediate host and are transmitted by predation to the same definitive host may cooperate in their attempts to manipulate the behaviour of the intermediate host, while at the same time having conflicts of interests over the use of host resources. A few studies have indicated that intermediate hosts harbouring larval helminths have altered concentrations of neurotransmitters in their nervous system, and thus measuring levels of neurotransmitters in host brains could serve to assess the respective and combined effect of different helminth species on host behaviour. Here, we investigate potential cooperation and conflict among three helminths in two species of crab intermediate hosts. The acanthocephalan Profilicollis spp., the trematode Maritrema sp. and an acuariid nematode, all use Macrophthalmus hirtipes (Ocypodidae) as intermediate host, whereas Profilicollis and Maritrema also use Hemigrapsus crenulatus (Grapsidae). All three helminths mature inside gulls or other shore birds. There was a significant decrease in the mean volume of Profilicollis cystacanths as the intensity of infection by this parasite increased in H. crenulatus, the only host in which this was investigated; however, there was no measurable effect of other helminth species on the size of acanthocephalans, suggesting no interspecific conflict over resource use within crabs. There was, in contrast, evidence of a positive interspecific association between the two most common helminth species: numbers of Profilicollis and Maritrema were positively correlated among crabs, independently of crab size, in M. hirtipes but not H. crenulatus. More importantly, we found that the total number of larval helminths per crab correlated significantly, and negatively, with concentrations of serotonin in crab brains, again only in M. hirtipes; numbers of each parasite species separately did not covary in either crab species with serotonin or dopamine, the other neurotransmitter investigated in this study. The relationship with serotonin appears due mainly to numbers of Profilicollis and Maritrema and not to nematodes. This is the first demonstration of a potentially synergistic manipulation of host behaviour by different helminth species, one that appears host-specific; our results also point toward the neurobiological mechanism underlying this phenomenon.     

Behavioral effects of serotonin and serotonin agonists in two crayfish species, Procambarus clarkii and Orconectes rusticus

Exogenous serotonin elicits several behaviors in Procambarus clarkii, including a flexed, elevated posture, reduced locomotion, and changes in aggressive behavior. We conducted experiments to determine if several serotonin agonists mimicked the behavioral effects of serotonin in two crayfish species, P. clarkii and Orconectes rusticus. Drugs tested were 1-(3-Chlorophenyl)-piperazine dihydrochloride (mCPP), Oxymetazoline, 2,5-dimethoxy-4-iodoamphetamine (DOI), CGS-12066A, and (+/-)-8-hydroxy-2-(di-n-dipropylamino) tetralin (8-OH-DPAT). In P. clarkii, mCPP most closely mimicked the effects of serotonin, significantly increasing the performance of the flexed, elevated posture and reducing locomotion; 8-OH-DPAT significantly reduced locomotion as well. Both of these drugs produced significant increases in elevated posture and decreases in locomotion in O. rusticus, and in this species, the drugs at test concentrations were more effective in eliciting these effects than serotonin. The effects of the drugs on behaviors performed during fighting bouts were variable. In both species, only 8-OH-DPAT significantly reduced several agonistic behaviors, and no agonist or 5-HT itself produced significant increases in agonistic behavior.     

Serotonin-immunoreactive neural system and contractile system in the hydroid Cladonema (Cnidaria, Hydrozoa)

Serotonin is a widespread neurotransmitter which is present in almost all animal phyla including lower metazoans such as Cnidaria. Serotonin detected in the polyps of several cnidarian species participates in the functioning of a neural system. It was suggested that serotonin coordinates polyp behavior. For example, serotonin may be involved in muscle contraction and/or cnidocyte discharge. However, the role of serotonin in cnidarians is not revealed completely yet. The aim of this study was to investigate the neural system of Cladonema radiatum polyps. We detected the net of serotonin-positive processes within the whole hydranth body using anti-serotonin antibodies. The hypostome and tentacles had denser neural net in comparison with the gastric region. Electron microscopy revealed muscle processes throughout the hydranth body. Neural processes with specific vesicles and neurotubules in their cytoplasm were also shown at an ultrastructural level. This work demonstrates the structure of serotonin-positive neural system and smooth muscle layer in C. radiatum hydranths.     

A-to-I editing of the 5HT2C receptor and behaviour.

Site-specific deamination of five adenosine residues in the pre-mRNA of the serotonin 2C receptor, 5HT2CR, alters the amino acid sequence of the encoded protein. Such RNA editing can produce 32 mRNA variants, encoding 24 protein isoforms that vary in biochemical and pharmacological properties. Because serotonin functions in the regulation of mood and behaviour, modulation of serotonin signalling by RNA editing may be relevant to such psychiatric disorders as anxiety and depression. Several recent human studies have reported changes in 5HT2CR editing in schizophrenia, major depression or suicide, but results are variable and not conclusive. Rodent studies have begun to examine effects of drug treatments and stress. Understanding the importance of 5HT2CR editing in mood and behaviour will be assisted by experiments designed to analyse multiple strains of mice, in different behavioural tests, with optimal evaluation of the time course of molecular changes.     

Effect of calcium channel blockers on serotonin uptake

We previously demonstrated that verapamil inhibits serotonin uptake by bovine pulmonary arterial endothelial cells by a mechanism not involving alterations in calcium fluxes. In this study, we determine whether verapamil inhibition of serotonin uptake occurs in other pulmonary cell types (bovine pulmonary artery smooth muscle cells), in cells from other organs and species (rat epididymal endothelial cells), and in intact organs (isolated rat lungs). We also compare the effects of verapamil with those of nifedipine and diltiazem. At concentrations of 10(-6) M or greater, verapamil is an inhibitor of serotonin uptake by cultured cells and isolated lungs. Nifedipine and diltiazem are weak inhibitors of serotonin uptake by cultured bovine cells only at suprapharmacologic doses and have no effect on serotonin uptake by isolated lungs. Surprisingly, nifedipine stimulates serotonin uptake by rat epididymal endothelial cells. We conclude that inhibition of serotonin uptake by verapamil is a generalized phenomenon, occurring in a variety of cell types, in intact organs, and in different species that does not occur consistently with other calcium channel blockers.     

Expression of synapsin and co-localization with serotonin and RFamide-like immunoreactivity in the nervous system of the chordoid larva of Symbion pandora (Cycliophora)

Abstract. Cycliophora is one of the most recently described metazoan phyla and hitherto includes only two species: Symbion pandora and Symbion americanus . With a very complex life cycle, cycliophorans are regarded as an enigmatic group with an uncertain phylogenetic position, although they are commonly considered lophotrochozoan protostomes. In order to extend the database concerning the distribution of immunoreactive substances in the free-swimming chordoid larva of S. pandora , we investigated synapsin immunoreactivity using fluorescence-coupled antibodies in combination with confocal laserscanning microscopy. Moreover, we analyzed the co-localization patterns of synapsin, serotonin, and RFamide-like immunoreactivity in the chordoid larva by 3D imaging technology based on the confocal microscopy image stacks. Synapsin is expressed in large parts of the bilobed anterior cerebral ganglion including anterior and dorsal projections. Two pairs of ventral neurites run longitudinally into the larval body of which the inner pair shows only weak, scattered synapsin immunoreactivity. In addition, a lateral synapsin immunoreactive projection emerges posteriorly from each ventral longitudinal axon. Double immunostaining shows co-localization of synapsin and serotonin in the cerebral ganglion, the outer and the inner ventral neurites, and the anterior projections. Synapsin and RFamide-like immunoreactivity co-occur in the cerebral ganglion, the outer ventral neurites, and the dorsal projections. Accordingly, the cerebral ganglion and the outer ventral neurites are the only neural structures that co-express the two neurotransmitters and synapsin. The overall neuroanatomical condition of the cycliophoran chordoid larva resembles much more the situation of adult rather than larval life cycle stages of a number of spiralian taxa.     

The developmental role of serotonin: news from mouse molecular genetics

New genetic models that target the serotonin system show that transient alterations in serotonin homeostasis cause permanent changes to adult behaviour and modify the fine wiring of brain connections. These findings have revived a long-standing interest in the developmental role of serotonin. Molecular genetic approaches are now showing us that different serotonin receptors, acting at different developmental stages, modulate different developmental processes such as neurogenesis, apoptosis, axon branching and dendritogenesis. Our understanding of the specification of the serotonergic phenotype is improving. In addition, studies have revealed that serotonergic traits are dissociable, as there are populations of neurons that contain serotonin but do not synthesize it.     

Cross-taxon congruence in the rarity of subtidal rocky marine assemblages: No taxonomic shortcut for conservation monitoring

The implementation of protection strategies such as the European Marine Strategy Framework Directive (2008/56/EC) is impeded for subtidal rock bottom habitats because of high sampling costs due to a very wide taxonomic diversity, and a lack of suitable evaluation tools to estimate their conservation importance. In this study, we seek to provide an evaluation procedure by (1) investigating the distribution of rarity among subtidal rock bottom phyla; (2) searching for potential surrogate phyla with a cross-taxon congruence approach based on their rarity; (3) proposing an appropriate multi-phyla indicator to evaluate the importance of subtidal rocky habitats for conservation. We analysed the distribution of 548 species belonging to 8 phyla sampled in 137 assemblages in subtidal rocky areas located around Brittany, Western France. We applied the Index of Relative Rarity, a flexible method which fits rarity weights to species depending on their respective phyla. We found only weak congruence in rarity patterns among phyla, which prevented any attempt to identify surrogate phyla. This finding has important implications for the conservation of subtidal rocky habitats as it means that there is no shortcut to monitor their rarity: working on a subset of phyla would imply a biased evaluation of biodiversity. Consequently, we propose a multi-phyla Index of Relative Rarity combining all phyla which allowed us to successfully describe rarity patterns across all sampled sites.     

Locomotor and peripheral effects of sibutramine modulated by 5-HT2 receptors

Sibutramine has been described as an anti-obesity drug with the ability to inhibit serotonin (5-HT), noradrenaline, and dopamine re-uptake, but without affinity to histamine and muscarinic receptors. On the other hand, cyproheptadine antagonizes serotonin 5-HT(2A), 5-HT(2B), and 5-HT(2C), histamine H1, and muscarinic (M) receptors. There are many reports concerning the influence of sibutramine on central serotoninergic pathways. In this study, we suggest that peripheral pathways may also be involved in the serotoninergic effects of sibutramine. In vivo experiments were undertaken to investigate the serotoninergic effects of sibutramine on body mass, the glycogen concentration in the diaphragm of rats, and locomotor behaviour. Rats were submitted to oral treatment with sibutramine, cyproheptadine, or sibutramine applied in combination with cyproheptadine, for a period of 2 months to investigate the 5-HT2 effects of sibutramine on these parameters. As the results demonstrated, the lower increase in body mass and the increased glycogen levels in the diaphragm muscle of rats treated with sibutramine seem to be modulated by 5-HT2 receptors, since these effects were completely antagonized by cyproheptadine in the group treated with the 2 drugs co-applied. Furthermore, the behavioural results also suggest that mechanisms modulated by 5-HT2 receptors are involved in the increase of locomotion in the rats treated with sibutramine, since the effect did not occur in the rats treated with sibutramine co-applied with the 5-HT2 receptor antagonist, cyproheptadine. The results suggest that sibutramine modifies energy-related parameters such as body mass, diaphragm glycogen, and locomotor behaviour in rats via 5-HT2 serotoninergic pathways.     

Serotonin modulates responses to species-specific vocalizations in the inferior colliculus

Neuromodulators such as serotonin are capable of altering the neural processing of stimuli across many sensory modalities. In the inferior colliculus, a major midbrain auditory gateway, serotonin alters the way that individual neurons respond to simple tone bursts and linear frequency modulated sweeps. The effects of serotonin are complex, and vary among neurons. How serotonin transforms the responses to spectrotemporally complex sounds of the type normally heard in natural settings has been poorly examined. To explore this issue further, the effects of iontophoretically applied serotonin on the responses of individual inferior colliculus neurons to a variety of recorded species-specific vocalizations were examined. These experiments were performed in the Mexican free-tailed bat, a species that uses a rich repertoire of vocalizations for the purposes of communication as well as echolocation. Serotonin frequently changed the number of recorded calls that were capable of evoking a response from individual neurons, sometimes increasing (15% of serotonin-responsive neurons), but usually decreasing (62% of serotonin-responsive neurons), this number. A functional consequence of these serotonin-evoked changes would be to change the population response to species-specific vocalizations.     

Insect allatotropins belong to a family of structurally-related myoactive peptides present in several invertebrate phyla

Originally named for its ability to stimulate juvenile hormone production by lepidopteran corpora allata, allatotropin has emerged as a neuropeptide with multiple neural, endocrine and myoactive roles. This paper describes the experimental evidence for allatotropin action, its localization in several species of insects, and its multiple effects on a variety of different tissues that lead to increased hemolymph circulation and gut motility. The overall physiological effects may also include species-specific effects such as the regulation of nutrient absorption, modulation of the circadian cycle and migratory preparedness. In addition, we present evidence suggesting that allatotropins are members of a family of myoactive peptides found in several invertebrate phyla. Finally, we speculate that the myoactive properties of allatotropins are basal and it is likely that the stimulatory action of allatotropins on juvenile hormone synthesis evolved secondarily.     

Serotonergic modulation of footshock induced aggression in paired rats.

Footshock induced aggression (FIA) was induced in paired rats and three paradigms of aggressive behaviour were recorded, namely, latency to fight (LF), total period of physical contact (TPP) and cumulative aggression scores (CAS). The effects of increasing or decreasing central serotonergic activity, by using a number of pharmacological agents with well defined effects on rat brain serotonin, were investigated on FIA and on FIA augmented by apomorphine, a dopamine receptor agonist. The results show that centrally administered serotonin, the serotonin precursor, 5-hydroxytryptophan administered with clorgyline, a selective MAO A inhibitor, quipazine, a serotonin receptor agonist, and fluoxetine, a selective inhibitor of neuronal re-uptake of serotonin, attenuated all paradigms of FIA and apomorphine induced potentiation of FIA. On the contrary, the other re-uptake inhibitor used, citalopram, appeared to have a dual effect and decreased LF and CAS, while increasing TPP. The serotonin synthesis inhibitor, p-chlorophenylalanine and the selective serotonin receptor (5-HT2) antagonist, ketanserin, augmented all paradigms of FIA per se and apomorphine induced augmentation of FIA. However, the other serotonin receptor antagonist used, metergoline, which blocks both 5-HT1 and 5-HT2 receptor subtypes, attenuated FIA per se but decreased only CAS in apomorphine induced increase in FIA. The data confirm the inhibitory effect of the central serotonergic system on aggressive behaviour and the inverse relationship existing between it and the central dopaminergic system in the modulation of FIA, as has also been confirmed in earlier biochemical investigations from this laboratory. The data has been discussed in the light of existing knowledge on serotonin receptor subtypes and the presence of modulatory serotonergic heteroreceptors on central dopaminergic neurones.