Morphological and functional alterations in human proximal tubular cell line induced by low level inorganic arsenic: evidence for targeting of mitochondria and initiated apoptosis.
The kidney is a known target organ for arsenic and is critical for both arsenic biotransformation and elimination. Previous studies have demonstrated that at high doses (ppm) inorganic arsenic is toxic to mitochondria primarily by affecting cellular respiration. However, the effect of inorganic arsenic on mitochondria after low level exposures is not known, particularly in the kidney. Thus the functional and morphological effects of low level inorganic arsenic were investigated in a human proximal tubular cell line, HK-2. Mitochondrial function was assessed at subcytotoxic concentrations of arsenite (< or = 10 microm) by examining the alteration of the mitochondrial membrane potential using MitoTracker Red, a mitochondrion selective dye. In a subset of cells, subcytotoxic arsenite led to mitochondrial membrane depolarization, which could subsequently lead to permeability transition and apoptosis. Subcytotoxic arsenite also induced translocation of phosphatidylserine, indicative of early-stage apoptosis. To confirm whether subcytotoxic arsenite induces cellular and/or mitochondrial morphological alterations consistent with initiated apoptosis, HK-2 cells were evaluated with transmission electron microscopy. Classic morphology of apoptosis was not observed with subcytotoxic arsenite exposures; however, evidence of necrotic changes in the cytoplasmic structure and mitochondrial morphology were apparent. Therefore, based on depolarization of mitochondria and the externalization of phosphatidylserine, HK-2 cells appear to initiate apoptosis following subcytotoxic arsenite insult, but morphological changes indicate that HK-2 cells fail to complete apoptosis and ultimately undergo necrosis. Therefore, subcytotoxic arsenite can be sufficiently toxic to mitochondria that they lose their ability to keep the cell on course for apoptotic cell death.
Peraza
Marjorie A
MA
Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona, Tucson, USA. map32@psu.edu
Cromey
Douglas W
DW
Carolus
Barb
B
Carter
Dean E
DE
Gandolfi
A Jay
AJ
D017209:Apoptosis; D001149:Arsenates / Q000633:toxicity; D018053:Arsenites / Q000633:toxicity*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D006801:Humans; D007687:Kidney Tubules, Proximal / Q000187:drug effects* / Q000648:ultrastructure; D008564:Membrane Potentials; D046529:Microscopy, Electron, Transmission; D008928:Mitochondria / Q000187:drug effects* / Q000648:ultrastructure; D051336:Mitochondrial Membranes / Q000187:drug effects; D009930:Organic Chemicals; D017670:Sodium Compounds / Q000633:toxicity*; D013997:Time Factors
D001149:Arsenates; D018053:Arsenites; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D017670:Sodium Compounds; C107472:red dye CMXRos; C017947:sodium arsenite; C009277:sodium arsenate