Journal Article

Cadmium-Induced Apoptosis in Primary Rat Cerebral Cortical Neurons Culture Is Mediated by a Calcium Signaling Pathway

Hu, Fei-fei, Liu, Xue-zhong, Jiang, Chen-yang, Sun, Ya, Liu, Zong-ping, Yuan, Yan, Bian, Jian-chun, Gu, Jian-hong, Xu, Hui

2013

Overview

Cadmium (Cd) is an extremely toxic metal, capable of severely damaging several organs, including the brain. Studies have shown that Cd disrupts intracellular free calcium ([Ca(2+)]i) homeostasis, leading to apoptosis in a variety of cells including primary murine neurons. Calcium is a ubiquitous intracellular ion which acts as a signaling mediator in numerous cellular processes including cell proliferation, differentiation, and survival/death. However, little is known about the role of calcium signaling in Cd-induced apoptosis in neuronal cells. Thus we investigated the role of calcium signaling in Cd-induced apoptosis in primary rat cerebral cortical neurons. Consistent with known toxic properties of Cd, exposure of cerebral cortical neurons to Cd caused morphological changes indicative of apoptosis and cell death. It also induced elevation of [Ca(2+)]i and inhibition of Na(+)/K(+)-ATPase and Ca(2+)/Mg(2+)-ATPase activities. This Cd-induced elevation of [Ca(2+)]i was suppressed by an IP3R inhibitor, 2-APB, suggesting that ER-regulated Ca(2+) is involved. In addition, we observed elevation of reactive oxygen species (ROS) levels, dysfunction of cytochrome oxidase subunits (COX-I/II/III), depletion of mitochondrial membrane potential (ΔΨm), and cleavage of caspase-9, caspase-3 and poly (ADP-ribose) polymerase (PARP) during Cd exposure. Z-VAD-fmk, a pan caspase inhibitor, partially prevented Cd-induced apoptosis and cell death. Interestingly, apoptosis, cell death and these cellular events induced by Cd were blocked by BAPTA-AM, a specific intracellular Ca(2+) chelator. Furthermore, western blot analysis revealed an up-regulated expression of Bcl-2 and down-regulated expression of Bax. However, these were not blocked by BAPTA-AM. Thus Cd toxicity is in part due to its disruption of intracellular Ca(2+) homeostasis, by compromising ATPases activities and ER-regulated Ca(2+), and this elevation in Ca(2+) triggers the activation of the Ca(2+)-mitochondria apoptotic signaling pathway. This study clarifies the signaling events underlying Cd neurotoxicity, and suggests that regulation of Cd-disrupted [Ca(2+)]i homeostasis may be a new strategy for prevention of Cd-induced neurodegenerative diseases.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Acetates - pharmacology / Adenosine diphosphate / Amino Acid Chloromethyl Ketones - pharmacology / Animals / Apoptosis / Apoptosis - drug effects / Bcl-2 protein / Biology / Boron Compounds - pharmacology / Brain damage / Ca(2+) Mg(2+)-ATPase - antagonists & inhibitors / Ca(2+) Mg(2+)-ATPase - genetics / Ca(2+) Mg(2+)-ATPase - metabolism / Ca2+/Mg2+-exchanging ATPase / Cadmium / Cadmium - pharmacology / Calcium / Calcium (intracellular) / Calcium - metabolism / Calcium homeostasis / Calcium ions / Calcium Signaling / Calcium signalling / Caspase / Caspase 3 - genetics / Caspase 3 - metabolism / Caspase 9 - genetics / Caspase 9 - metabolism / Caspase-3 / Caspase-9 / Cell culture / Cell death / Cell proliferation / Cell survival / Cerebral Cortex - cytology / Cerebral Cortex - drug effects / Cerebral Cortex - metabolism / Cytochrome / Egtazic Acid - analogs & derivatives / Egtazic Acid - pharmacology / Electron Transport Complex IV - genetics / Electron Transport Complex IV - metabolism / Elevation / Exposure / Fetus / Gene Expression Regulation / Homeostasis / Inhibitors / Inositol 1,4,5-trisphosphate receptors / Intracellular / Kinases / Mathematics / Medicine / Membrane potential / Mitochondria / Mortality / Neurodegenerative diseases / Neurological diseases / Neurons / Neurons - cytology / Neurons - drug effects / Neurons - metabolism / Neurotoxicity / Organs / Oxygen / Poly(ADP-ribose) polymerase / Poly(ADP-ribose) Polymerases - genetics / Poly(ADP-ribose) Polymerases - metabolism / Primary Cell Culture / Proto-Oncogene Proteins c-bcl-2 - genetics / Proto-Oncogene Proteins c-bcl-2 - metabolism / Rats / Rats, Sprague-Dawley / Reactive oxygen species / Reactive Oxygen Species - metabolism / Ribose / Rodents / Signal transduction / Sodium-Potassium-Exchanging ATPase - antagonists & inhibitors / Sodium-Potassium-Exchanging ATPase - genetics / Sodium-Potassium-Exchanging ATPase - metabolism / Studies / Toxicity / Veterinary colleges