Keys molecules implicated in apoptotic pathway of sea urchin embryos

Sea urchin embryos cadmium exposed as an experimental model system for studying the relationship between autophagy and apoptosis

Cadmium accumulation induces apoptosis in sea urchin embryos and larvae

Cadmium induces an apoptotic response in sea urchin embryos.

Cadmium is a heavy metal toxic for living organisms even at low concentrations. It does not have any biological role, and since it is a permanent metal ion, it is accumulated by many organisms. In the present paper we have studied the apoptotic effects of continuous exposure to subacute/sublethal cadmium concentrations on a model system: Paracentrotus lividus embryos. We demonstrated, by atomic absorption spectrometry, that the intracellular amount of metal increased during exposure time. We found, using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay, that long treatments with cadmium triggered a severe DNA fragmentation. We demonstrated, by immunocytochemistry …

Environmental factors as possible causes of DNA fragmentation in human sperms

Literature data demonstrated that some environmental factors could have a key role in the remarkable and continuous decline of sperm quality observed in the last fifty years. Specifically, in the Taranto area, data about the detrimental effects of environmental pollution are alarming because of the high level of poisons released in the atmosphere. Pollution coming from the plant causes health and fertility risks, mainly due to the exposure to the dioxin. Our study analyzed sperm samples from three patients groups: i) workers of local steel factories; ii) Taranto residents; iii) Controls. Results demonstrate that patients from the “factory workers” group, constantly exposed to environmental …

Cadmium effects on sea urchin embryos

Apoptotic pathway induced by cadmium in sea urchin embryos

MITOCHONDRIA DURING SEA URCHIN OVOGENESIS

Autophagy is related to apoptosis in sea urchin embryos exposed to cadmium

The sea urchin embryo is a suitable model system that offers an excellent opportunity to investigate different defense strategies activated in stress conditions. We previously showed that cadmium treatment provokes the accumulation of metal in dose-time dependent manner in embryonic cells and the activation of defense systems, such as the synthesis of HSPs and/or the initiation of apoptosis. Analyzing autophagy, by neutral red, acridine orange and LC3-detection, we demonstrated that Cd-exposed embryos adopt this process as an additional stratagem to safeguard the developmental program. We observed that embryos treated with subletal Cd concentration activate massive autophagic response after…

Autophagy is related to apoptosis in Paracentrotus lividus embryos cadmium exposed

P. lividus embryo offers an excellent opportunity to investigate the adaptive response of cells exposed to different stress. We previously demonstrated that cadmium treatment triggers the accumulation of metal in embryonic cells and the activation of defense system depending on concentration and exposure time, through the synthesis of HSPs and/or the initiation of apoptosis. Analysing autophagy, by neutral red, acridine orange and LC3-detection, we demonstrated that Cd-exposed embryos adopt this process as an additional stratagem to safeguard the developmental program. We observed that embryos treated at subletal Cd concentration activate a massive autophagic response after 18h, which decre…

A method for measuring mitochondrial mass and activity

Mitochondria, responsible for the energy-generating process essential for the cell metabolism, differ for the number, localization and activity in animal cells and tissues in relation to the energetic needs. Using fluorescent probes specific for mitochondria, Mitotracker Green (MTG) and Orange (MTO), and Confocal Laser-Scanning Microscope (CLSM), we elaborated a method to measure in vivo the mitochondrial mass and activity, in sea urchin Paracentrotus lividus eggs and embryos. The analysis of captured images, revealed a variation of mitochondrial distribution and an increase of activity after fertilization.

Autophagy is required for sea urchin oogenesis and early development.

SummaryAutophagy is a major intracellular pathway for the degradation and recycling of cytosolic components. Emerging evidence has demonstrated its crucial role during the embryo development of invertebrates and vertebrates. We recently demonstrated a massive activation of autophagy in Paracentrotus lividus embryos under cadmium stress conditions, and the existence of a temporal relationship between induced autophagy and apoptosis. Although there have been numerous studies on the role of autophagy in the development of different organisms, information on the autophagic process during oogenesis or at the start of development in marine invertebrates is very limited. Here we report our recent …

Cadmium insult and defence mechanisms in Paracentrotus lividus embryos and larvae.

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

It is well known that sea urchin embryos are able to activate different defense strategies against stress. We previously demonstrated that cadmium treatment triggers the accumulation of metal in embryonic cells and the activation of defense systems depending on concentration and exposure time, through the synthesis of heat shock proteins and/or the initiation of apoptosis. Here we show that Paracentrotus lividus embryos exposed to Cd adopt autophagy as an additional stratagem to safeguard the developmental program. At present, there are no data focusing on the role of this process in embryo development of marine organisms. In this paper we utilized different techniques to detect autophagy i…

Cadmium accumulation induces apoptosis in P. lividus larvae.

Apoptosis during early development of sea urchin.

Apoptosis is a genetic program of cell death that eliminates superfluous or compromised cells during development and adult life of many organisms. In sea urchin embryos, apoptosis is not only a physiological event during larval metamorphosis, but also a process induced by cadmium accumulation or other stressor like TPA (12-O-tetradecanoylphorbol-13-acetate) followed by an increase of temperature to 31°C. Apoptosis is a highly conserved process usually operated by a proteolytic cascade that involves caspase activation by two different pathways: extrinsic and intrinsic. The first one involves membrane death receptors, while the second involves mitochondria. In this work we analyzed the possib…

APOPTOTIC ANALYSIS OF CUMULUS CELLS FOR THE SELECTION OF COMPETENT OOCYTES TO BE FERTILIZED BY INTRACYTOPLASMIC SPERM INJECTION (ICSI)

Oocyte quality is one of the main factors for the success of in vitro fertilization protocols. Apoptosis is known to affect oocyte quality and may impair subsequent embryonic development and implantation. The aim of this study was to investigate the apoptosis rate of single and pooled cumulus cells of cumulus cell–oocyte complexes (COCs), as markers of oocyte quality, prior to intracytoplasmatic sperm injection (ICSI).We investigated the apoptosis rate by TUNEL assay (DNA fragmentation) and caspase-3 immunoassay of single and pooled cumulus cells of COCs. The results showed that DNA fragmentation in cumulus cells was remarkably lower in patients who achieved a pregnancy than in those who di…

Induction of apoptosis by cadmium in Paracentrotus lividus embryos and larvae

Induction of apoptosis by cadmium in sea urchin embryos: possible involvement of extrinsic pathway

L’autofagia durante l’oogenesi e i primi stadi di sviluppo embrionale in Paracentrotus lividus

Autophagy during development of sea urchin embryos

Autophagy is a highly regulated mechanism that enhances cell eukaryotic survival under various environmental and cellular stresses, by breakdown and recycling of macromolecules and organelles. Here we report that in Paracentrotus lividus embryos autophagic process occur, at a lesser extend during physiological development and at greater levels after a cadmium treatment. By Acridine Orange staining, we found that embryonic cells exposed to cadmium display green fluorescence in cytoplasm and nucleus, and show considerable red fluorescent dots in cytoplasm. This evidence suggests formation of acidic auto-phagolysosomal vacuoles. By Neutral Red vital staining, specific for lysosomes and cellula…

The autophagic demand during oogenesis and early development of sea urchin

The autophagic pathway is an evolutionarily conserved homeostatic process, responsible for degradation and recycling of long-term proteins and cytoplasmic organelles in eukaryotic cells. This process constitutively occurs at basal levels and is involved in cell survival. Increased autophagy is induced by environmental cues, such as starvation and many stress agents, while excessive levels of autophagy can lead to autophagic Programmed Cell Death, with features that differ from those of the apoptotic process. We recently demonstrated massive activation of autophagy in P. lividus embryos, in cadmium stress conditions, and the existence of a temporal relationship between induced autophagy end …

Effect of environmental stress on sea urchin embryos and larvae: from developmental to molecular biology analyses

Response to cadmium stress in sea urchin embryos

Hsp56 expression in Paracentrotus lividus embryos

Apoptosis: focus on sea urchin development

It has been proposed that the apoptosis is an essential requirement for the evolution of all animals, in fact the apoptotic program is highly conserved from nematodes to mammals. Throughout development, apoptosis is employed by multicellular organisms to eliminate damaged or unnecessary cells. Here, we will discuss both developmental programmed cell death (PCD) under normal conditions and stress induced apoptosis, in sea urchin embryos. Sea urchin represent an excellent model system for studying embryogenesis and cellular processes involved in metamorphosis. PCD plays an essential role in sculpting and remodelling the embryos and larvae undergoing metamorphosis. Moreover, this marine organi…

Sea urchin embryo as a model system for studing autophagy induced by cadmium stress

The Role of Autophagy and Apoptosis During Embryo Development

Programmed cell death (PCD) and cell survival are two sides of the same coin. Autopha‐ gy and apoptosis are crucial processes during embryo development of Invertebrates and Vertebrates organisms, as they are necessary for the formation of a new organism, start‐ ing from a fertilized egg. Fertilization triggers cell remodeling from each gamete to a toti‐ potent zygote. During embryogenesis, the cells undergo various processes, thus allowing the transformation of the embryo into an adult organism. In particular, cells require the appropriate tools to suddenly modify their morphology and protein content in order to respond to intrinsic and external stimuli. Autophagy and apoptosis are involved…

Localization of mitochondrial Hsp56 chaperonin during sea urchin development.

We have previously demonstrated that Paracentrotus lividus nuclear genome encodes for the heat shock inducible chaperonin homolog Hsp 56 (1) and that the mature protein is localized in the mitochondrial matrix (2). In this paper we report that constitutive Hsp56 is maternally inherited, in fact it is present in the in unfertilized eggs, and that it has a perinuclear specific localization during cleavage. In the later stages both the constitutive and the heat shock inducible chaperonin has a specific territorial distribution. Moreover following heat shock, the Hsp56 appears in the cytoplasm and in the postmitochondrial supernatant beside the mitochondrial fraction.

L'embrione di riccio di mare come modello di studio dell'autofagia indotta da stress

Gli embrioni di riccio di mare, Paracentrotus lividus, sono in grado di attivare differenti strategie di difesa come risposta a stress chimico/fisici. Recentemente abbiamo dimostrato che il cadmio, metallo pesante altamente embriotossico, induce la sintesi di specifiche hsps e/o l’innesco di processi apoptotici, via via che si accumula nelle cellule embrionali. Nel presente lavoro, mostriamo che gli embrioni di P. lividus sono in grado di attivare l’autofagia come un aggiuntivo meccanismo atto a salvaguardare il programma di sviluppo, in seguito a esposizione a dosi citotossiche di CdCl2. L’autofagia è un meccanismo molecolare che può stimolare la sopravvivenza, attraverso la degradazione e…

Cadmium induces autophagy during development of Paracentrotus lividus embryos.

Mitochondria during sea urchin oogenesis.

SummarySea urchin represents an ideal model for studies on fertilization and early development, but the achievement of egg competence and mitochondrial behaviour during oogenesis remain to be enlightened. Oocytes of echinoid, such as sea urchin, unlike other echinoderms and other systems, complete meiotic maturation before fertilization. Mitochondria, the powerhouse of eukaryotic cells, contain a multi-copy of the maternally inherited genome, and are involved directly at several levels in the reproductive processes, as their functional status influences the quality of oocytes and contributes to fertilization and embryogenesis. In the present paper, we report our latest data on mitochondrial…

Risposta da stress in embrioni di Paracentrotus lividus esposti a cadmio

Autophagy

Klionsky, Daniel J. et al.

Physiological and cadmium induced apoptosis in Paracentrotus lividus larvae

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

Seuls les 100 premiers auteurs dont les auteurs INRA ont été entrés dans la notice. La liste complète des auteurs et de leurs affiliations est accessible sur la publication.; International audience; In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues…

Effetti del cadmio sugli embrioni di riccio di mare

Apoptosis rate of cumulus cells can be considered as an indicator for the selection of embryos to improve ongoing pregnancy and implantation rates.

Cumulus cells apoptosis rate an adjunct to morphology evaluation for the embryo selection on day 3 could be considered a new tool, compared with embryo selection by morphology alone, to select the embryos with higher implantation potential to increase the clinical outcomes after ICSI. Several studies have demonstrated a lower cumulus cell apoptotic rates in women who achieved pregnancy compared with women who did not become pregnant after ICSI. A prospective randomized observational study on 76 ICSI patients was performed before Ovum Pick-Up. Patients were randomized into either the control group (embryo selection by morphology only, A group: 48 patients) or the treatment group (morphology …

Confocal microscopy study of the distribution, content and activity of mitochondria during Paracentrotus lividus development

Summary In the present paper we applied confocal microscopy andfluorescence technologies for studying the distribution andthe oxidative activity of sea urchin ( Paracentrotus lividus )mitochondria during development, by in vivo incubating eggsand embryos with cell-permeant MitoTracker probes. Wecalculated, by a mathematical model, the intensity values, the variations of intensity, and the variation index of incorporatedfluorochromes. Data demonstrate that mitochondrial massdoes not change during development, whereas mitochondrialrespirationincreases.Inaddition,startingfrom16blastomeresstage, some regions of the embryo contain organelles moreactive in oxygen consumption. Introduction The con…

A possible role of autophagy for the execution of apoptosis in cadmium-exposed sea urchin embryos

The sea urchin embryo is a suitable model system that offers an excellent opportunity to investigate different defence strategies activated in stress conditions. We previously showed that cadmium treatment provokes the accumulation of metal in dose- and time-dependent manner in embryonic cells and the activation of defence systems, such as the synthesis of HSPs and/or the initiation of apoptosis. Analysing autophagy, by neutral red, acridine orange and LC3-detection, we demonstrated that Cd-exposed embryos adopt this process as an additional stratagem to safeguard the developmental program. We observed that embryos treated with subletal Cd concentration activate a massive autophagic respons…

Differences beetween physiological and induced apoptosis in sea urchin embryos.

P. lividus oogenesis and early development require autophagy

It is well known that autophagy is a major intracellular pathway for the degradation and recycling of cytosolic components, in both basal and stress conditions. We have recently demonstrated the activation of autophagy in P. lividus embryos: at high levels, after a stress induced by cadmium, and at basal levels, during the physiological development (1) (2). Here we report ours recent data about autophagy during oogenesis and segmentation. In order to detect autophagolysosomes (AVOs) and autolysosomes, we respectively performed incubation in vivo with acridine orange (AO), and in situ immunofluorescence (IF) with the anti-LC3 antibody (autophagy marker). All observations were carried out by …

p38 MAPK in cadmium-treated MDA-MB231 breast cancer cells

Stress induced and physiological apoptosis during early development of sea urchin Paracentrotus lividus.

Cadmium effects on p38/MAPK isoforms in MDA-MB231 breast cancer cells

Emerging evidence seems to indicate that the heavy metal cadmium (Cd) is able to regulate gene expression, drastically affecting the pattern of transcriptional activity in normal and pathological eukaryotic cells, also affecting intracellular signalization events. Human p38 is a family of mitogen-activated protein kinases consisting of four isoforms (alpha, beta, gamma and delta) which mediate signal transduction cascades controlling several aspects of cell physiology. In this study we examined whether exposure of MDA-MB231 tumor cells from the human breast to Cd may exert some effect on p38 isoform expression and accumulation, as well as on p38 activation. Employing a combination of prolif…

Autophagy as defence strategy against cadmium stress in Paracentrotus lividus embryos.

Sea urchin embryo is a developmental model that offers an excellent opportunity to investigate the possible adaptive response of cells exposed to different stress during differentiation. These embryos are able to respond to many stress by synthesizing a set of highly conserved proteins, the hsps and/or by activation of apoptosis. The exposure to cadmium trigger the accumulation of metal in embryo cells and the activation of both defence mechanisms depending on concentration and exposure time [1-4]. Recent experimental evidences demonstrate that by autophagy, a highly regulated mechanism that enhances cell survival under various environmental and cellular stress, the breakdown and recycling …

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

Autophagy is used by organisms as a defense strategy to face environmental stress. This mechanism has been described as one of the most important intracellular pathways responsible for the degradation and recycling of proteins and organelles. It can act as a cell survival mechanism if the cellular damage is not too extensive or as a cell death mechanism if the damage/stress is irreversible; in the latter case, it can operate as an independent pathway or together with the apoptotic one. In this review, we discuss the autophagic process activated in several aquatic organisms exposed to different types of environmental stressors, focusing on the sea urchin embryo, a suitable system recently in…

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition) 1

Contains fulltext : 232759.pdf (Publisher’s version ) (Closed access) In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to…

Relationship between autophagy and apoptosis in Paracentrotus lividus embryos cadmium exposed

Cadmium is a strong toxicant for living organism since it does not have biological roles and can cause several cellular damages comprising genotoxicity, oxidative stress, and other biochemical dysfunctions. Marine invertebrate embryos represent a suitable model system where to investigate the effects of many stressors on development and cell viability. Here we investigated the toxic effect of cadmium on sea urchin Paracentrotus lividus embryos focusing our attention on metal-inducead autophagy and the possible temporal and functional relationship with apoptosis. Using several techniques to detect autophagy (neutral red, acridine orange and LC3-detection) we demonstrated that Cd-exposed P. l…

Apoptosi indotta da cadmio in embrioni di riccio di mare

Autophagy

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide…

Cadmium induces apoptosis in P. lividus embryos

Ruolo dell’autofagia nell’oogenesi di riccio di mare Paracentrotus lividus

Erratum

Author(s): Klionsky, DJ; Abdelmohsen, K; Abe, A; Abedin, MJ; Abeliovich, H; Arozena, AA; Adachi, H; Adams, CM; Adams, PD; Adeli, K; Adhihetty, PJ; Adler, SG; Agam, G; Agarwal, R; Aghi, MK; Agnello, M; Agostinis, P; Aguilar, PV; Aguirre-Ghiso, J; Airoldi, EM; Ait-Si-Ali, S; Akematsu, T; Akporiaye, ET; Al-Rubeai, M; Albaiceta, GM; Albanese, C; Albani, D; Albert, ML; Aldudo, J; Algul, H; Alirezaei, M; Alloza, I; Almasan, A; Almonte-Beceril, M; Alnemri, ES; Alonso, C; Altan-Bonnet, N; Altieri, DC; Alvarez, S; Alvarez-Erviti, L; Alves, S; Amadoro, G; Amano, A; Amantini, C; Ambrosio, S; Amelio, I; Amer, AO; Amessou, M; Amon, A; An, Z; Anania, FA; Andersen, SU; Andley, UP; Andreadi, CK; Andrieu-Ab…

Relazione tra autofagia e apoptosi in embrioni di riccio di mare esposti a stress

L’embrione di riccio di mare Paracentrotus lividus è stato utilizzato come organismo modello della biologia dello sviluppo per molti anni ed è considerato il deuterostoma più primitivo, con scheletro calcificato, correlato a protocordati e vertebrati. Tale sistema offre un'eccellente opportunità per studiare le numerose strategie di difesa che gli embrioni mettono in atto contro diverse condizioni di stress. In precedenza, abbiamo riportato che l’esposizione di embrioni a dosi citotossiche di cadmio, provoca l'accumulo intracellulare del metallo e l'attivazione del sistema difensivo, in modo dose-tempo dipendente, attraverso la sintesi di specifiche HSPs e/o l’innesco di apoptosi. Mediante …

Apoptotic pathway in sea urchin embryos

Cytotoxic effects induced by JA47, a novel histone deacetylase inhibitor (HDACi), on MDA-MB231 breast cancer cells

Sea urchin embryos exposed to cadmium as an experimental model for studying the relationship between autophagy and apoptosis

The sea urchin embryo is a suitable model that offers an excellent opportunity to investigate different defence strategies activated in stress conditions. We previously showed that cadmium accumulates in a dose- and time-dependent manner into embryonic cells, activating different stress and defence mechanisms, including the synthesis of HSPs and the onset of apoptosis and/or autophagy. In this paper we investigated the functional relationship between autophagy and apoptosis, evaluating apoptosis signals in cadmium-exposed Paracentrotus lividus embryos with inhibited autophagy. We found that the inhibition of autophagy produced the concurrent reduction of apoptosis, suggesting that the two p…

Cadmium induces the expression of specific stress proteins in sea urchin embryos.

Abstract Marine organisms are highly sensitive to many environmental stresses, and consequently, the analysis of their bio-molecular responses to different stress agents is very important for the understanding of putative repair mechanisms. Sea urchin embryos represent a simple though significant model system to test how specific stress can simultaneously affect development and protein expression. Here, we used Paracentrotus lividus sea urchin embryos to study the effects of time-dependent continuous exposure to subacute/sublethal cadmium concentrations. We found that, between 15 and 24 h of exposure, the synthesis of a specific set of stress proteins (90, 72–70, 56, 28, and 25 kDa) was ind…

Apoptosi in larve di Paracentrotus lividus.