Early appearance and possible functions of non-neuromuscular cholinesterase activities

The biological function of the cholinesterase (ChE) enzymes has been studied since the beginning of the twentieth century. Acetylcholinesterase plays a key role in the modulation of neuromuscular impulse transmission in vertebrates, while in invertebrates pseudo cholinesterases are preeminently represented. During the last 40 years, awareness of the role of ChEs role in regulating non-neuromuscular cell-to-cell interactions has been increasing such as the ones occurring during gamete interaction and embryonic development. Moreover, ChE activities are responsible for other relevant biological events, including regulation of the balance between cell proliferation and cell death, as well as the modulation of cell adhesion and cell migration. Understanding the mechanisms of the regulation of these events can help us foresee the possible impact of neurotoxic substances on the environmental and human health

Adhesion of sea-urchin living cells on nano-patterned anodic porous alumina

In this work we investigated the possibility of using living cells as stress sensing material in biosensors, in the light of the three Rs principle \u2013 Replacement, Reduction and Refinement . This approach requires the necessity to cultivate them on biocompatible electrical conducting substrate and to insert the circuit into a culture chamber that must assure both the transport of oxygen and the diffusion of the medium containing the potential stressor to the cells, without modifying their response and the structure of the culture. To this aim we fabricated nano-patterned substrates of anodic porous alumina to be used for enhancing cell adhesion, and culture chambers made in polylactic acid. Sea-urchin cells (coelomocytes) were cultured on these substrates at different times of 1, 3 and 5 days in vitro. Since these cells are progenitors of immune cells in vertebrate systems (blood cells), they carry out similar functions. For this reason, although they can differ considerably from vertebrates, they have been proved to be very promising sentinels of environmental water qualit

Cholinergic Pesticides

Neurotoxicity exerted by organophosphates and carbamate

Supramolecular Organization of the Repetitive Backbone Unit of the Streptococcus pneumoniae Pilus

Streptococcus pneumoniae, like many other Gram-positive bacteria, assembles long filamentous pili on their surface through which they adhere to host cells. Pneumococcal pili are formed by a backbone, consisting of the repetition of the major component RrgB, and two accessory proteins (RrgA and RrgC). Here we reconstruct by transmission electron microscopy and single particle image reconstruction method the three dimensional arrangement of two neighbouring RrgB molecules, which represent the minimal repetitive structural domain of the native pilus. The crystal structure of the D2-D4 domains of RrgB was solved at 1.6 Å resolution. Rigid-body fitting of the X-ray coordinates into the electron density map enabled us to define the arrangement of the backbone subunits into the S. pneumoniae native pilus. The quantitative fitting provide evidence that the pneumococcal pilus consists uniquely of RrgB monomers assembled in a head-to-tail organization. The presence of short intra-subunit linker regions connecting neighbouring domains provides the molecular basis for the intrinsic pilus flexibility

Localization of some neurotransmitters during developments hydroidomedusae

Neurotransmission in Cnidarians is known to occur mainly by peptidergic neuronal pathways; the role of cholinergic neurotransmission, that in other animals this involves both neurotransmission and developmental regulation, has never been shown with certainty. This histochemical-immunohistochemical study was performed, in different developmental stages of some hydroidomedusan species (characterized by different reproductive and developmental patterns) to investigate the presence and distribution of cholinesterase (ChE) activity, acetylcholine receptors, biogenic amines, and molecules immunologically related to secretory peptides (FMRF amide and VIP). The results showed that ChE (revealed by all the kinds of activities: AChE, PrChE, BuChE) is mainly present in developing or 'juvenile' structures. Biogenic amines are revealed by aldehyde-induced fluorescence at every stage studied, while peptide-like immunoreactivities are mainly present in differentiated structures. These findings support the hypothesis that all the chemical messengers here investigated arose very early in evolution, although, in some cases, with different functions. For most of the investigated species, similarities in the distribution pattern of markers generally reflect similarities in life cycle, rather than phylogenetic affinities. © 1994.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Muscarinic signalling affects intracellular calcium concentration during the first cell cycle of sea urchin embryos.

The existence of a response to acetylcholine (ACh) and cholinomimetic drugs in sea urchin eggs and zygotes was investigated in two sea urchin species: Paracentrotus lividus and Lytechinus pictus. The calcium sensitive fluorescent probe, Fura-2 dextran, was employed to investigate the regulation of cytosolic free calcium concentration ([Ca(2+)](i)) by cholinomimetic drugs in unfertilised and fertilised eggs of both the sea urchin species. Exposure to cholinomimetic agonists/antagonists, either extracellularly or intracellularly, had no effect either on resting [Ca(2+)](i) levels in the unfertilised sea urchin egg, or on the transient [Ca(2+)](i) increase at fertilisation. However, following fertilisation, extracellular application of ACh receptors agonists, such as ACh and carbachol, predominantly muscarinic agonist, but not nicotine, induced a significant increase in [Ca(2+)](i), which was partially inhibited by atropine. As a consequence of exposure after fertilisation to the agonists of ACh receptors, chromatin structure was transiently affected. The hypothesis is proposed that muscarinic receptors may be involved in the (presumably Ca(2+)-dependent) modulation of the nuclear status during the first cell cycles

Micronuclei induction and neurotoxic effect in C6 glioma cells exposed to low concentrations of diazinon, an organophosphorus compound

Abstract Background: The presence of doses of diazinon far lower than IC50 cholinesterase activity was reported in plasma of pregnant women and newborns living in agricultural areas. Objective: In the current study, we investigated the possibility of neurotoxicity induction by exposing cultured gliotypic C6 cells to a similar range of concentrations, for 24 h, at 37\ub0C. Materials and methods: Confluent C6 cellswere exposed to diazinon (DZN) at concentrations from 200ng/L to 0.002ng/L. The maintenance of confluence, the induction of micronuclei and the expression of molecules related to the cholinergic system were verified, by morphological, biochemical and immunohistochemical methods, in order to check the effects of the altered modulation of the cholinergic signal on glial-like cells. Results: The exposure to 0.002ng/L showed significant effecton micronuclei occurrence since the exposure to 0.002ng/L, while the inhibition on butyrylcholinesterase activity showed significant variations starting from the exposure to 0.2ng/L up to 200ng/L. Acetylcholinesterase activity was significantly inhibited only by the exposure to 200ng/L. The immunohistochemical localization of choline acetyltransferase and fibronectin showed dramatic variation only in C6 cells exposed to 200ng/L. Conclusion: The low doses of DZN investigated affect the investigated features of glial-like cells, mainly starting from the 0.2ng/L dose, while the effects on AChE activity and ChAT and fibronectin-immuorectivity were clearly exerted in cell cultures exposed to 200ng/L. Collectively, these findings translated to the in vivo functions of glial cells indicate that exposure to doses that are nontoxic to adult organisms may weaken the brain defense and functions of glial cells through an AChE-mediated mechanism

Reproductive biology of Parazoanthus axinellae (Schmidt, 1862) and Savalia savaglia (Bertoloni, 1819) (Cnidaria, Zoantharia) from the NW Mediterranean coast

Despite the wide distribution of zoanthids, little is known about their pattern of reproduction. Here we investigate the reproductive biology of two Mediterranean species, the common Parazoanthus axinellae (Schmidt) and the rare Savalia savaglia (Bertoloni). For both species, samples were collected during an annual cycle, from January to December 2005, in the Western Mediterranean (Ligurian Sea, Italy). Both species are gonochoric. In P. axinellae the sex-ratio (n colonies = 30) showed a slight predominance of male colonies (M/F = 1.35), whereas in the population of S. savaglia (n colonies = 15) a predominance of females was found (M/F = 0.3). In P. axinellae the first gametocytes were visible in March, whereas in S. savaglia they became visible in May. Both species reproduce at the end of autumn when seawater temperature begins to decrease. Parazoanthus axinellae (10 m depth) spawns eggs and sperms in November, whereas S. savaglia (67 m depth) spawns in December. In P. axinellae sexes were segregated on a rocky wall, with males occurring deeper, whereas male and female colonies of S. savaglia were irregularly dispersed in the population. The maximum number of oocytes differed between the species, being higher in P. axinellae than in S. savaglia