Use of flow cytometry and confocal microscopy techniques to investigate early CdCl2-induced nephrotoxicity in vitro

Alvarez-Barrientos, Alberto; O’Connor, José Enrique; Castillo, R; Moreno, Alfonso Blázquez; Prieto, Pilar

doi:10.1016/s0887-2333(01)00044-3

Cited by 27 publications

(18 citation statements)

References 8 publications

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“…Our data is in line with other tissue culture studies using cadmium at micromolar ranges (Alvarez-Barrientos et al 2001;Bagchi et al 2000;Chao and Yang 2001;Dong et al 2001a;Hader et al1996). We have also investigated the effect of exposing the embryos to 1 lM cadmium for 24 h. The percentage of apoptotic cells, as determined by the flow cytometry assay, was 0.67±0.06%.…”

Section: Discussionsupporting

confidence: 93%

“…Murine thymocytes treated with 5-50 lM cadmium for 24 h have a significantly higher percentage of sub-G 1 apoptotic cells when compared with the control (Dong et al 2001a). Similarly, apoptosis was evident only after 24 h of exposure to 25 and 100 lM cadmium in the dog distal tubule cell line MDCK and the pig proximal tubule cell line LLC-PK1 (Alvarez- Barrientos et al 2001). …”

Section: Introductionmentioning

confidence: 99%

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Cadmium-induced ectopic apoptosis in zebrafish embryos

2003

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In this study, we tested the hypothesis that cadmium-induced developmental toxicity was mediated via ectopic occurrence of apoptosis during embryonic development. We employed confocal microscopy to acquire images of whole-mount staining of apoptotic cells in zebrafish embryo exposed to 100 micro M cadmium from 5 hours post fertilisation (hpf) to 28 hpf. Three-dimensional reconstruction of the images was performed and the spatial and temporal distributions of apoptotic cells in the embryos were compared. In cadmium-treated embryos with varying degrees of gross developmental malformations, significantly higher numbers of apoptotic cells were detected with this method. In order to detect the precise locations of apoptotic cells, we performed terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL) assay in sectioned embryos. In the degenerating neural tube of cadmium-treated embryos apoptotic cells were detected, while in the healthy neural tube of the untreated controls no apoptotic cells were found. We then employed flow cytometry to investigate whether cadmium exposure would affect the dynamics of apoptosis or induce any abnormalities in cell-cycle progression. It appeared that cadmium did not induce cell-cycle arrest. The percentages of apoptotic cells did not differ in the two groups at 13, 16 or 19 hpf. At 28 hpf, however, a significantly higher percentage of apoptotic cells were found in the cadmium-treated group. Exposure to cadmium, therefore, induced ectopic apoptosis at 28 hpf without affecting the dynamics of apoptosis at earlier developmental stages.

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Section: Discussionsupporting

confidence: 93%

Section: Introductionmentioning

confidence: 99%

Cadmium-induced ectopic apoptosis in zebrafish embryos

2003

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“…Moreover, proximal renal tubule epithelial cells are rich in mitochondria, and mitochondrial dysfunction seems to be involved in several types of xenobiotic-induced nephrotoxicity [23]. Herein, renal cortex of rats was collected to investigate the mitochondrial damage when administered with lead and/or cadmium for 8 weeks.…”

Section: Discussionmentioning

confidence: 99%

Effects of Lead and/or Cadmium on the Oxidative Damage of Rat Kidney Cortex Mitochondria

Lin

et al. 2009

Biol Trace Elem Res

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Lead acetate (300 mg/L) and/or cadmium chloride (50 mg/L) were administered as drinking water to Sprague-Dawley rats for 8 weeks to investigate the possible combined effects of these metals on the damage in renal cortex mitochondria. Increased malonaldehyde levels due to exposure to these metals were detected by colorimetric method, which demonstrated the lipid peroxidation in the renal cortex. Ultrastructural observations and real-time quantitative PCR analyses were performed on kidney cortex pieces to identify the mitochondrial damage and quantify the relative expression levels of cytochrome oxidase subunits (COX-I/II/III), respectively. The most striking ultrastructural modifications involved distortion of mitochondrial cristae and an unusual arrangement, which were more evident when the mixture was ingested. There were significant differences in the expression levels of COX-I, II, and III between the control group and the exposed groups, whereas those in the (lead+cadmium) group were all significantly lower than that in the lead or cadmium group. In conclusion, there was an obvious synergistic oxidative damage effect of lead combined with cadmium on rat kidney cortex mitochondria, which increased defects in mitochondrial oxidative metabolism.

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“…Concerning the mechanism of Cd-induced renal damage, several lines of evidence indicate that oxidative stress and apoptotic death formed in the presence of cadmium could be responsible for its toxic effects [4][5][6][7][8][9][10]. Cytosolic Cd generates ROS, which deplete endogenous radical ROS scavengers and damage a variety of transport proteins, including inhibition of the redox-sensitive enzymes and membrane ion-motive ATPases [5,6].…”

Section: Introductionmentioning

confidence: 99%

“…If these ROS-mediated stress events are not balanced by repair processes, affected cells undergo apoptosis or necrosis [7]. Also, mitochondrial dysfunction seems to be involved in the Cd-induced nephrotoxicity due to the fact that proximal renal tubule epithelial cells are rich in mitochondria [8]. In vitro, cadmium can induce apoptosis directly through targeting of mitochondria at low micromolar concentrations [9].…”

Section: Introductionmentioning

confidence: 99%

Role of Oxidative Stress, Apoptosis, and Intracellular Homeostasis in Primary Cultures of Rat Proximal Tubular Cells Exposed to Cadmium

Wang

Cao

Chen

et al. 2008

Biol Trace Elem Res

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Cadmium (Cd) is a known nephrotoxic element. In this study, the primary cultures of rat proximal tubular (rPT) cells were treated with low doses of cadmium acetate (2.5 and 5 microM) to investigate its cytotoxic mechanism. A progressive loss in cell viability, together with a significant increase in the number of apoptotic and necrotic cells, were seen in the experiment. Simultaneously, elevation of intracellular [Ca(2+)]i and reactive oxygen species (ROS) levels, significant depletion of mitochondrial membrane potential(Delta Psi) and cellular glutathione (GSH), intracellular acidification, and inhibition of Na(+), K(+)-ATPase and Ca(2+)-ATPase activities were revealed in a dose-dependent manner during the exposure, while the cellular death and the apoptosis could be markedly reversed by N-acetyl-L-cysteine (NAC). Also, the calcium overload and GSH depletion were significantly affected by NAC. In conclusion, exposure of rPT cells to low-dose cadmium led to cellular death, mediated by an apoptotic and a necrotic mechanism. The apoptotic death might be the chief mechanism, which may be mediated by oxidative stress. Also, a disorder of intracellular homeostasis induced by oxidative stress and mitochondrial dysfunction is a trigger of apoptosis in rPT cells.

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Use of flow cytometry and confocal microscopy techniques to investigate early CdCl2-induced nephrotoxicity in vitro

Cited by 27 publications

References 8 publications

Cadmium-induced ectopic apoptosis in zebrafish embryos

Cadmium-induced ectopic apoptosis in zebrafish embryos

Effects of Lead and/or Cadmium on the Oxidative Damage of Rat Kidney Cortex Mitochondria

Role of Oxidative Stress, Apoptosis, and Intracellular Homeostasis in Primary Cultures of Rat Proximal Tubular Cells Exposed to Cadmium

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