Assessment of the molluscicidal impact of extracted chlorophyllin on some biochemical parameters in the nervous tissue and histological changes in Biomphalaria alexandrina and Lymnaea natalensis snails Abstract Biomphalaria alexandrina and Lymnaea natalensis snails are the intermediate hosts of schistosomiasis and fasciolosis. The aim of the present study is to evaluate the molluscicidal activity of chlorophyll extract as a photodynamic substance against these snails and how it affected its tissues and the biological system. Chlorophyllin was extracted from deep-frozen Moringa oleifera leaves, and then it was transformed into water-soluble chlorophyllin. The present results showed that it had a molluscicidal activity on B. alexandrina snails (LC50 17.6 mg/l; LC90 20.9 mg/l) and L. natalensis snails (LC50 4.3 mg/l; LC90 6.8 mg/l). Exposing B. alexandrina snails to the sublethal concentrations (LC0, LC10, and LC25) resulted in a significant reduction in their survival rates. Regarding its effect on biochemical parameters, chlorophyllin significantly reduced the acetylcholinesterase activity, protein content, and alkaline and acid phosphatase activity in B. alexandrina nervous tissue compared to the control group. Histopathological changes occurred in the digestive gland of treated B. alexandrina snails where cells lost their nuclei, vacuolated, degenerated, and ruptured, and the lumen increased. Photosynthesizing materials like chlorophyllin are new approaches to control schistosomiasis and fasciolosis in developing countries by affecting their intermediate host. These materials were cheap and environmentally safe to replace the synthetic molluscicides for snail control.

Identification of putative amine biosynthetic enzymes in the nervous system of the crab, Cancer borealis Abstract Amines function as neuromodulators throughout the animal kingdom. In decapod crustaceans, the amines serving neuromodulatory roles include dopamine, octopamine, serotonin and histamine. While much work has focused on examining the physiological effects of amines on decapod nervous systems, the identity of the native enzymes involved in their biosynthesis remains largely unknown. In an attempt to help fill this void, a transcriptome generated from multiple portions of the crab, Cancer borealis, nervous system, a species that has long served as a model species for investigating the neuromodulatory control of rhythmically active neural networks, was used to identify putative amine biosynthetic enzyme-encoding transcripts, and by proxy, proteins. Transcripts encoding full complements of the enzymes involved in the production of dopamine, octopamine, serotonin, and histamine were deduced from the C. borealis assembly, i.e., tryptophan–phenylalanine hydroxylase, tyrosine hydroxylase, DOPA decarboxylase, tyrosine decarboxylase, tyramine β-hydroxylase, tryptophan hydroxylase, and histidine decarboxylase. All proteins deduced from the C. borealis transcripts appear to be full-length sequences, with reciprocal BLAST and structural domain analyses supporting the protein family annotations ascribed to them. These data provide the first descriptions of the native amine biosynthetic enzymes of C. borealis, and as such, serve as a resource for initiating gene-based studies of aminergic control of physiology and behavior at the level of biosynthesis in this important biomedical model.

Neural pathways in the pallial nerve and arm nerve cord revealed by neurobiotin backfilling in the cephalopod mollusk Octopus vulgaris Abstract Here, we report the findings after application of neurobiotin tracing to pallial and stellar nerves in the mantle of the cephalopod mollusk Octopus vulgaris and to the axial nerve cord in its arm. Neurobiotin backfilling is a known technique in other molluscs, but it is applied to octopus for the first time to be best of our knowledge. Different neural tracing techniques have been carried out in cephalopods to study the intricate neural connectivity of their nervous system, but mapping the nervous connections in this taxon is still incomplete, mainly due to the absence of a reliable tracing method allowing whole-mount imaging. In our experiments, neurobiotin backfilling allowed: (1) imaging of large/thick samples (larger than 2 mm) through optical clearing; (2) additional application of immunohistochemistry on the backfilled tissues, allowing identification of neural structures by coupling of a specific antibody. This work opens a series of future studies aimed to the identification of the neural diagram and connectome of octopus nervous system.

Lidocaine and carbamazepine inhibit while phenytoin and lamotrigine paradoxically enhance the insect neuromuscular transmission Abstract Primary mechanism of action of local anesthetics and various anticonvulsants is the voltage-gated sodium channel block. Many of these small molecules also have other targets in nervous system of vertebrates. However, little is known about their action on invertebrate nervous system. Nevertheless, insect-based models are suggested for high-throughput screening of antiepileptic drugs. In the present work, we characterized action of lidocaine, carbamazepine, lamotrigine, and phenytoin on the neuromuscular transition of Calliphora vicina fly larvae using conventional voltage-clamp approach. Carbamazepine and lidocaine caused inhibition of synaptic transmission, which has presynaptic origin. This action is in agreement with inhibition of voltage-gated sodium channels that reduces depolarization of nerve terminals and, thus, calcium entry. Surprisingly, phenytoin and lamotrigine produced a prominent increase in the evoked postsynaptic currents without any effect on frequency or amplitude of spontaneous miniature currents. Potassium channel blocker 4-aminopyridine affects synaptic transmission in similar way. Elevation of synaptic quantal content via increase in calcium concentration or via application of 1 mM 4-aminopyridine eliminates the enhancement effect or even turns it to modest inhibition. We propose that lamotrigine and phenytoin act as inhibitors of insect potassium channels that cause the membrane depolarization and thus facilitates calcium entry into the nerve terminal.

Dysregulation of axogenesis in the antennal nervous system of the embryonic grasshopper Schistocerca gregaria Abstract The antennal nervous system of the grasshopper Schistocerca gregaria features two parallel axon tracts each established early in embryogenesis by discrete pairs of pioneer neurons located at the antennal tip and whose growth cones contact so-called base pioneers en route to the brain. Here we present two antennal phenotypes in which a stereotypic dysregulation of axogenesis in a given tract is observed when only the base pioneer associated with that pathway is missing, consistent with a role for this cell type in guided axogenesis. Dysregulation involves defasciculation and aberrant navigation by pioneer axons resulting in a missing or depleted primordial antennal nerve to the brain. The dysregulated phenotypes reveal that axogenesis in each pathway is regulated independently. Previously unseen discrepancies in the navigational decisions made by pioneer neurons which derive sequentially from the same mother cell demonstrate that these progeny have separate identities. Possible mechanisms for the dysregulated phenotypes are considered.

A study of the role of vision in the foraging behaviour of the pyrrhocorid bug Antilochus conquebertii (Insecta; Hemiptera; Pyrrhocoridae) Abstract Our study aims to describe (1) external morphology of the compound eye of Antilochus conquebertii, (2) postembryonic changes involving the eye’s shape and size and (3) behaviour of the animal with respect to the organization of the compound eye. With each moult of the insect, the structural units of the compound eye increase in size as well as the number, resulting in an overall increase in eye size. The resolution of the adult eye is better than the young one. The adult possesses UV and polarization sensitivity in its eye. Parallel to the changes of the eye the behaviour of the adult animal changes, rendering it increasingly nocturnal and less active in under illuminated conditions. The current study describes the eye and its functional relationship with the behaviour of the animal at the nymphal and adult developmental stage. Graphical abstract

Na + /K + -pump and neurotransmitter membrane receptors Abstract Na+/K+-pump is an electrogenic transmembrane ATPase located in the outer plasma membrane of cells. The Na+/K+-ATPase pumps 3 sodium ions out of cells while pumping 2 potassium ions into cells. Both cations move against their concentration gradients. This enzyme’s electrogenic nature means that it has a chronic role in stabilizing the resting membrane potential of the cell, in regulating the cell volume and in the signal transduction of the cell. This review will mainly consider the role of the Na+/K+-pump in neurons, with an emphasis on its role in modulating neurotransmitter receptor. Most of the literature on the modulation of neurotransmitter receptors refers to the situation in the mammalian nervous system, but the position is likely to be similar in most, if not all, invertebrate nervous systems.

Yeast two-hybrid screening identifies MPZ-1 and PTP-1 as candidate scaffolding proteins of metabotropic glutamate receptors in Caenorhabditis elegans Abstract The metabotropic glutamate receptors (mGluRs) are a class of G-protein-coupled receptor that undergo extensive interactions with scaffolding proteins, and this is intrinsic to their function as an important group of neuromodulators at glutamatergic synapses. The Caenorhabditis elegans nervous system expresses three metabotropic glutamate receptors, MGL-1, MGL-2 and MGL-3. Relatively little is known about how the function and signalling of these receptors is organised in C. elegans. To identify proteins that scaffold the MGL-1 receptor, we have conducted a yeast two-hybrid screen. Three of the interacting proteins, MPZ-1, NRFL-1 and PTP-1, displayed motifs characteristic of mammalian mGluR scaffolding proteins. Using cellular co-expression criterion, we show mpz-1 and ptp-1 exhibited overlapping expression patterns with subsets of mgl-1 neurons. This included neurones in the pharyngeal nervous system that control the feeding organ of the worm. The mGluR agonist L-CCG-I inhibits the activity of this network in wild-type worms, in an MGL-1 and dose-dependent manner. We utilised L-CCG-I to identify if MGL-1 function was disrupted in mutants with deletions in the mpz-1 gene. The mpz-1 mutants displayed a largely wild-type response to L-CCG-I, suggesting MGL-1 signalling is not overtly disrupted consistent with a non-obligatory modulatory function in receptor scaffolding. The selectivity of the protein interactions and overlapping expression identified here warrant further investigation of the functional significance of scaffolding of metabotropic glutamate receptor function.

A rapid introduction to neurological biochemistry using Drosophila melanogaster Abstract Short, cost-effective teaching activities are a useful way of providing an integrated view on biological processes. Here we describe a brief, hands-on workshop that allows pre-university students to explore their understanding of a neurological pathway from its chemical bases to phenotype. The workshop effectively introduces the students to data collection and analysis in an enjoyable way and at an appropriate level, determined by an end of session feedback survey. The design of the workshop can be adapted and scaled to generate diverse sessions such as university teaching practicals or summer school training workshops.

Investigation of feeding behaviour in C. elegans reveals distinct pharmacological and antibacterial effects of nicotine Abstract Caenorhabditis elegans is an informative model to study the neural basis of feeding. A useful paradigm is one in which adult nematodes feed on a bacterial lawn which has been pre-loaded with pharmacological agents and the effect on pharyngeal pumping rate scored. A crucial aspect of this assay is the availability of good quality bacteria to stimulate pumping to maximal levels. A potential confound is the possibility that the pharmacological agent impacts bacterial viability and indirectly influences feeding rate. Here, the actions of nicotine on pharyngeal pumping of C. elegans and on the Escherichia coli bacterial food source were investigated. Nicotine caused an immediate and concentration-dependent inhibition of C. elegans pharyngeal pumping, IC50 4 mM (95% CI = 3.4 mM to 4.8 mM). At concentrations between 5 and 25 mM, nicotine also affected the growth and viability of E. coli lawns. To test whether this food depletion by nicotine caused the reduced pumping, we modified the experimental paradigm. We investigated pharyngeal pumping stimulated by 10 mM 5-HT, a food ‘mimic’, before testing if nicotine still inhibited this behaviour. The IC50 for nicotine in these assays was 2.9 mM (95% CI = 3.1 mM to 5.1 mM) indicating the depletion of food lawn does not underpin the potency of nicotine at inhibiting feeding. These studies show that the inhibitory effect of nicotine on C. elegans pharyngeal pumping is mediated by a direct effect rather than by its poorly reported bactericidal actions.