Aluminium Induced DNA-damage and Oxidative Stress in Cultures of the Marine Sponge Hymeniacidon perlevis

Abstract

Aluminium is the most abundant element in the earth crust, and has no known biological function. However, it is an established neurotoxicant in its trivalent oxidation state, with exposure resulting in neurodegenerative diseases like Parkinson’s disease and presenile dementia. Although, the potential genotoxic and carcinogenic effects of aluminium are established in mammalian and other model system, there is however very limited information on aluminium genotoxicity in aquatic invertebrates. Mechanism of aluminium toxicity is also largely unclear. With a concentration range between 0.001– 0.05mg/L in near neutral pH water, and up to 0.5-1mg/L in an acidic water , aluminium poses a potential threat to the marine ecosystem, however it is poorly studied. This study, therefore presents for the first time, aluminium-induced DNA damage using the comet assay and reactive oxygen Species (ROS) formation using 2’, 7’-dichlorodihydrofluorescein diacetate (H2DCF-DA) assay as biomarkers of genotoxicity and oxidative stress in the inter-tidal marine sponge Hymeniacidon perlevis, respectively. H. perlevis is widely distributed in the British Isles, Mediterranean and the Arctic sea and has been reported as a model for environmental biomonitoring in aquatic ecosystem and as a suitable alternative to bivalves. In this study, cryopreserved single sponge cells of H. perlevis were cultured as viable aggregates and were thereafter treated with 0.1, 0.2, 0.3 and 0.4mg/L aluminium chloride (AlCl3) for 12 hours. Cell viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Our results showed that non-cytotoxic concentrations of AlCl3 caused a statistically significant concentration-dependent increase in the level of DNA-strand break and reactive oxygen species formation single sponge cells of H. perlevis. There was also a statistically significant positive linear correlation between aluminium-induced DNA strand break and ROS formation suggesting the involvement of ROS in the causative mechanism of the aluminium induced DNA-strand breaks observed.

A broad‐taxa approach as an important concept in ecotoxicological studies and pollution monitoring

“…for testing various endpoints under controlled laboratory and field experiments; (ii) the possibility of assessing the impact of various pollutants on regeneration (Best et al, 1981;Best & Morita, 1982) using new models like artificial embryos (Johnson et al, 1982;Johnson & Gabel, 1983), the nubbins assay (Shafir et al, 2003) and the use of primmorphs (Akpiri et al, 2020) that include endpoints such as the inhibition of regeneration, regeneration time and formation of teratomas; (iii) evaluation of the direct effects of pollutants on stem cell populations, and the use of these systems to design effective interpretations of in vitro outcomes in order to understand whole-body impact better; and (iv) investigation of the epigenetically mediated inheritance of the impacts of pollution by unexposed descendants (Ellis, Kissane & Lynch, 2021b;Rosner et al, 2021).…”

“…Additionally, the extensive asexual reproduction and regeneration capabilities of sponges, even from adult body fragments (Baldacconi et al ., 2010; Çelik et al ., 2011), has enabled the production of clonal animals providing reproducible responses to contamination (Osinga, Tramper & Wijffels, 1999; Schippers et al ., 2012) that can be used for pollution monitoring. Furthermore, primmorphs, artificial models generated from sponges belonging to the taxa Demospongiae, Calcarea, and Homoscleromorpha (Lavrov & Kosevich, 2014; Akpiri, Konya & Hodges, 2020), also have potential as in vitro tools for toxicological tests. Primmorphs are obtained from sponge tissues dissociated into cells by mechanical or chemical methods.…”

“…Many hitherto underused species (poriferans, cnidarians, planarians, and colonial ascidians) have exceptional characteristics such as enormous regenerative ability, and the possession of populations of pluripotent ASCs, capable of differentiation into both soma and germ lineages (Rinkevich et al ., 2022; Rosner et al ., 2021; Ballarin et al ., 2022). These features can be exploited in the field of (eco)toxicology in many ways: ( i ) production of many ramets from a specific genet, all with identical genetic information and epigenomes, for testing various endpoints under controlled laboratory and field experiments; ( ii ) the possibility of assessing the impact of various pollutants on regeneration (Best et al ., 1981; Best & Morita, 1982) using new models like artificial embryos (Johnson et al ., 1982; Johnson & Gabel, 1983), the nubbins assay (Shafir et al ., 2003) and the use of primmorphs (Akpiri et al ., 2020) that include endpoints such as the inhibition of regeneration, regeneration time and formation of teratomas; ( iii ) evaluation of the direct effects of pollutants on stem cell populations, and the use of these systems to design effective interpretations of in vitro outcomes in order to understand whole‐body impact better; and ( iv ) investigation of the epigenetically mediated inheritance of the impacts of pollution by unexposed descendants (Ellis, Kissane & Lynch, 2021 b ; Rosner et al ., 2021).…”