Sea urchin embryos as a model system for studying autophagy induced by cadmium stress

Abstract

In this paper we utilized different techniques to detect autophagy in sea urchin embryos. Using neutral red (NR) and acridine orange (AO) vital staining, we found that embryos exposed to Cd display massive punctiform spots in the cytoplasm, indicative of acidic vesicular organelles (AVOs). These results have been confirmed using bafilomycin A 1 , a known inhibitor of this process. Furthermore these data were validated through both protein gel blotting and immunofluorescence in situ analysis of LC3-II, a specific marker of autophagy.We found that in P. lividus embryos autophagic processes occur in lesser amounts during physiological development and in greater amounts after Cd exposure.

Autophagy as a defense strategy against stress: focus on Paracentrotus lividus sea urchin embryos exposed to cadmium

“…Autophagy has been reported for the first time in Paracentrotus lividus embryos in 2011, and it is now known as one of the most important cellular/molecular pathways triggered by sea urchin embryos both in physiological and stress conditions (Chiarelli et al 2011). Recently, some genes involved in the regulation of autophagy were identified in Strongylocentrotus purpuratus (KEGG PATHWAY: spu04140).…”

“…These researches indicated that the exposure of the developing embryo to subacute/sublethal concentrations of Cd induce, firstly, the activation of the early defense mechanisms, such as the synthesis of HSPs and metallothioneins (Russo et al 2003;Roccheri et al 2004), but if this is not sufficient to safeguard the developmental program, embryos will trigger the autophagic and/or apoptotic processes (Chiarelli et al 2011;Agnello et al 2007). Some of the pathways activated in sea urchin P. lividus embryos, showed in Table 1, can be occur in tandem or in sequence.…”

“…In this case, second hypothesis, autophagy could provide the energy necessary for apoptosis through the supply of ATP molecules, by recycling of damaged cellular components. According to the third hypothesis, since the sea urchin embryo does not have specialized phagocytes, it could begin the clearance of apoptotic bodies through autophagy (Chiarelli et al 2011).…”

“…Due to its sensitivity to several aquatic contaminants, the sea urchin embryo adopts different defense mechanisms to preserve the developmental program (Chiarelli et al 2011). We will discuss the relationship between the activation of autophagy and/or apoptosis: a temporal and functional choice that depends on the intensity of the stress conditions, suggesting a choice made in a hierarchical way.…”

Cytotoxic Effects of Jay Amin Hydroxamic Acid (JAHA), a Ferrocene-Based Class I Histone Deacetylase Inhibitor, on Triple-Negative MDA-MB231 Breast Cancer Cells

“…Electrophoretic analysis and immunoblots were performed according to Chiarelli et al 21 Essentially, control and JAHA-treated cell lysates obtained after 18, 24, and 48 h of culture were submitted to 8% SDS-PAGE and protein transfer onto Hybond-ECL nitrocellulose membranes (Amersham) using a semidry apparatus (Novablot, Pharmacia). Immunorevelation was performed with either of the two primary antibodies, that is, rabbit polyclonal anti-LC3 (L8918, Sigma, St. Louis, MO; 1:1000) or anti-p62/SQSTM1 (P0068, Sigma; 1:1000), and antiactin antibodies (A5060, Sigma; 1:500) as a loading control and secondary alkaline phosphatase-conjugated antirabbit IgG (Fc) antibody (S373B, Promega, Madison, WI), followed by incubation with 5-bromo-4-chloro-3′-indolyphosphate p-toluidine salt/nitro-blue tetrazolium chloride (BCIP/NBT) solution (Sigma).…”