Whole-mount in situ TUNEL method revealed ectopic pattern of apoptosis in cadmium treated naupliar larvae of barnacle (Balanus amphitrite Darwin)

Cheng, Shuk Han; Chan, Kwok Wing; Chan, Pui-Kwong; So, Chun Hung; Wu, Rudolf S.S.

doi:10.1016/j.chemosphere.2004.01.011

Cited by 8 publications

(5 citation statements)

References 33 publications

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“…In green, DNA fragmentation (a, d, g, j); in red, nucleic acids (propidium iodide; b, e, h, k); merges of green and red (c, f, i, l). Larvae treated with low (d-f), medium (g-i), and high (j-l) cadmium; control larvae (a-c); Bar=100 μm concomitant increase in cadmium concentration (Cheng et al 2004). In conclusion, despite their high resistance to chemicals during their embryonic life described by other authors (Hamdoun and Epel 2007), sea urchin larvae continuously exposed to cadmium from fertilization, accumulate abnormalities during time.…”

Section: Discussionmentioning

confidence: 99%

Environmentally relevant cadmium concentrations affect development and induce apoptosis of Paracentrotus lividus larvae cultured in vitro

et al. 2008

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Sea urchin embryos and larvae represent suitable model systems on where to investigate the effects of heavy metals on development and cell viability. Here, we tested the toxic effects of low (10(-12 )M), medium (10(-9 )M), and high (10(-6 )M) cadmium chloride concentrations, mimicking unpolluted, moderately and highly polluted seawaters, respectively, on Paracentrotus lividus sea urchins offspring. Larvae were continuously treated from fertilization and inspected at time intervals comprised between 10 and 30 days of development. Delays and/or morphological abnormalities were firstly evident in larvae treated for 15 days with high cadmium (10(-6 )M) and for 25 days with medium cadmium (10(-9 )M). Major defects consisted in the reduction and lack of arms and skeleton elongation. No obvious differences with respect to controls were observed in embryos/larvae exposed to low cadmium (10(-12) M), even after 30 days of exposure. Using in situ terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay (TUNEL) assay on larvae whole mounts, we detected apoptosis after 10 days of treatment with 10(-6) and 10(-9) M CdCl(2,) when no morphological abnormalities were recognizable yet. Supernumerary apoptotic cells were found in arm buds, ciliary bands, and apex. In conclusion, echinoderm embryos and larvae represent candidates of choice for the study of stress and defense mechanisms activated by cadmium exposure.

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Section: Discussionmentioning

confidence: 99%

Environmentally relevant cadmium concentrations affect development and induce apoptosis of Paracentrotus lividus larvae cultured in vitro

et al. 2008

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“…Benthic invertebrates are commonly affected, since many of these organic pollutants have a substantial affinity for sedimentary absorption [103]. Similarly, heavy metals like manganese (liberated from MnO 2 during hypoxia; [104]) and cadmium [105] are frequent contaminants in marine sediments. Cadmium is perhaps the most common apoptosis-inducing environmental pollutant [105].…”

Section: Positive Stimuli For Apoptosismentioning

confidence: 99%

“…Similarly, heavy metals like manganese (liberated from MnO 2 during hypoxia; [104]) and cadmium [105] are frequent contaminants in marine sediments. Cadmium is perhaps the most common apoptosis-inducing environmental pollutant [105]. We offer here a brief synopsis of representative work that illustrates induction of apoptosis by these agents in crustaceans, but unfortunately there is paucity of information addressing the underlying mechanisms of signal transduction.…”

Section: Positive Stimuli For Apoptosismentioning

confidence: 99%

“…To investigate the loss of motility and survivorship, Cheng, et al [105] examined the apoptotic effects of cadmium exposure on stage II nauplius larvae of the barnacle, Balanus amphitrie . Using the TUNEL assay, these authors determined that the rate of apoptosis increased as the concentration of cadmium was raised to 15 μM, compared to untreated controls that lacked detectable apoptotic cells.…”

Section: Positive Stimuli For Apoptosismentioning

confidence: 99%

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Mechanisms of apoptosis in Crustacea: what conditions induce versus suppress cell death?

et al. 2009

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Arthropoda is the largest of all animal phyla and includes about 90% of extant species. Our knowledge about regulation of apoptosis in this phylum is largely based on findings for the fruit fly Drosophila melanogaster. Recent work with crustaceans shows that apoptotic proteins, and presumably mechanisms of cell death regulation, are more diverse in arthropods than appreciated based solely on the excellent work with fruit flies. Crustacean homologs exist for many major proteins in the apoptotic networks of mammals and D. melanogaster, but integration of these proteins into the physiology and pathophysiology of crustaceans is far from complete. Whether apoptosis in crustaceans is mainly transcriptionally regulated as in D. melanogaster (e.g., RHG ‘killer’ proteins), or rather is controlled by pro- and anti-apoptotic Bcl-2 family proteins as in vertebrates needs to be clarified. Some phenomena like the calcium-induced opening of the mitochondrial permeability transition pore (MPTP) are apparently lacking in crustaceans and may represent a vertebrate invention. We speculate that differences in regulation of the intrinsic pathway of crustacean apoptosis might represent a prerequisite for some species to survive harsh environmental insults. Pro-apoptotic stimuli described for crustaceans include UV radiation, environmental toxins, and a diatom-produced chemical that promotes apoptosis in offspring of a copepod. Mechanisms that serve to depress apoptosis include the inhibition of caspase activity by high potassium in energetically healthy cells, alterations in nucleotide abundance during energy-limited states like diapause and anoxia, resistance to opening of the calcium-induced MPTP, and viral accommodation during persistent viral infection. Characterization of the players, pathways, and their significance in the core machinery of crustacean apoptosis is revealing new insights for the field of cell death.

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“…Increased or induced apoptotic events have been reported in a wide range of aquatic species including Mytilus galloprovincialis exposed to tributlytin (Micic et al, 2001) and copper (Steinert, 1996); zebrafish embryos exposed to heat, UV, c-irradiation (Yabu et al, 2001), and aquatic hypoxia (Shang & Wu, 2004); barnacle larvae exposed to cadmium (Cheng et al, 2004); and Bufo arenarum tadpoles treated with cypermethrin (Casco et al, 2006). This study is focused on detecting apoptotic cells in the embryonic and early larval stages of the two invasive bivalves, the Mediterranean mussel (Mytilus galloprovincialis) (Lamarck) and the zebra mussel (Dreissena polymorpha) (Pallas) using the TUNEL assay as a terminal indicator.…”

Section: Introductionmentioning

confidence: 98%

Detecting physiological and pesticide-induced apoptosis in early developmental stages of invasive bivalves

Perry

Lynn

2009

Hydrobiologia

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In this study, embryos and early larval stages of two invasive bivalves, the Mediterranean mussel (Mytilus galloprovincialis) and zebra mussel (Dreissena polymorpha), were assayed for physiological apoptosis and stress-induced apoptosis post-exposure to a molluscicide, Bayluscide Ò . Physiological apoptosis was measured at 4-h intervals and Bayluscide Ò exposures were 4 h and 24 h beginning at both 30 min post-fertilization and at 24 h postfertilization. Apoptosis was detected by the TUNEL assay, and apoptotic values were compared for the different stages present. The two bivalves tested for apoptotic cells displayed variations in physiological and stress-induced apoptotic values despite having the same life stage transitions. However, in the majority of exposures, we found that as pesticide concentrations increased, the apoptotic values decreased in the highest concentrations. We suggest that this change to apoptotic inhibition indicates that there is a threshold at which apoptosis can no longer be used as a protective measure against damaged cells. These concentrations that cause changes in apoptotic pattern should be further tested for longterm effects on reproduction and survival. By detecting sub-lethal concentrations that may impair recruitment to the population and the continuation of the life cycle, chemical control of invasive bivalves could be targeted to early life stages and to reduce potential environmental impacts resulting from the higher concentrations required to control adults.

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Whole-mount in situ TUNEL method revealed ectopic pattern of apoptosis in cadmium treated naupliar larvae of barnacle (Balanus amphitrite Darwin)

Cited by 8 publications

References 33 publications

Environmentally relevant cadmium concentrations affect development and induce apoptosis of Paracentrotus lividus larvae cultured in vitro

Environmentally relevant cadmium concentrations affect development and induce apoptosis of Paracentrotus lividus larvae cultured in vitro

Mechanisms of apoptosis in Crustacea: what conditions induce versus suppress cell death?

Detecting physiological and pesticide-induced apoptosis in early developmental stages of invasive bivalves

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