Collapse of mitochondrial membrane potential and caspases activation are early events in okadaic acid-treated Caco-2 cells.
Diarrhetic Shellfish Poisoning (DSP) results from the consumption of shellfish contaminated with okadaic acid (OA) or one of the dinophysistoxins (DTX). It has been reported that this toxin induces apoptosis in several cell models, but the molecular events involved in this process have not been clarified. In this report we studied intracellular signals induced by OA in Caco-2 cells: mitochondrial membrane potential, F-actin depolymerization, caspases activation, cell proliferation and cell membrane integrity. Results indicate that caspases-8 and -9 increased their activity after 30 min of OA treatment according to their role as initiator caspases. In contrast, activation of the downstream caspase-3 is a later event in the execution phase of apoptosis. Mitochondrial membrane potential changes are detected at 30 min of OA exposure indicating that this is an early response in the apoptotic cascade. F-actin depolymerization occurs after 24h of incubation with OA and this effect is significant at low doses of the toxin. LDH is released into the culture medium, although there is not PI uptake, indicative of a significant cell death in addition to apoptosis. Moreover, OA led to a dose- and time-dependent decrease in cellular proliferation.
Lago
Jorge
J
Microbiology and Biotoxins Area, ANFACO-CECOPESCA, Campus Univ Vigo, 36310 Vigo (Pontevedra), Spain.
Santaclara
Francisco
F
Vieites
Juan M
JM
Cabado
Ana G
AG
D000199:Actins / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects; D018938:Caco-2 Cells; D020169:Caspases / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects; D004789:Enzyme Activation / Q000187:drug effects; D006801:Humans; D007202:Indicators and Reagents; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects*; D019319:Okadaic Acid / Q000633:toxicity*; D011419:Propidium / Q000378:metabolism
D000199:Actins; D007202:Indicators and Reagents; D019319:Okadaic Acid; D011419:Propidium; D007770:L-Lactate Dehydrogenase; D020169:Caspases