In vitro cell-toxicity screening as an alternative animal model  for coral toxicology: effects of heat stress, sulfide, rotenone,  cyanide, and cuprous oxide on cell viability and mitochondrial  function.
The logistics involved in obtaining and maintaining large numbers of corals hampers research on the toxicological effects of environmental contaminants for this ecologically and economically important taxon. A method for creating and culturing single-cell suspensions of viable coral cells was developed. Cell segregation/separation was based on specific cell densities and resulting cell cultures were viable for at least 2 mos. Low-density cells lacking symbiotic zooxanthallae and rich in mitochondria were isolated and cultured for toxicity studies. Cells were exposed to differing degrees or concentrations of heat stress, rotenone, cyanide, sulfide, and cuprous oxide. Cells were assayed for mitochondrial membrane potential using the fluorescent probe, JC-9, and for overall viability using the MTT/formazan spectrophotometric viability assay. Significant differences were observed between controls and treatments and the efficacy of this method was validated; only 2 cm(2) of tissue was required for a seven-point concentration-exposure series.
Downs
Craig A
CA
Haereticus Environmental Laboratory, P.O. Box 92, Clifford, VA 24533, USA. cadowns@gmail.com
Fauth
John E
JE
Downs
Virgil D
VD
Ostrander
Gary K
GK
D000704:Analysis of Variance; D000818:Animals; D037421:Anthozoa / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003300:Copper / Q000633:toxicity; D004785:Environmental Pollutants / Q000633:toxicity*; D005456:Fluorescent Dyes; D005562:Formazans; D006358:Hot Temperature; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D023421:Models, Animal; D011190:Potassium Cyanide / Q000633:toxicity; D012402:Rotenone / Q000633:toxicity; D013440:Sulfides / Q000633:toxicity; D013778:Tetrazolium Salts; D018675:Toxicity Tests / Q000379:methods
D004785:Environmental Pollutants; D005456:Fluorescent Dyes; D005562:Formazans; D013440:Sulfides; D013778:Tetrazolium Salts; D012402:Rotenone; C079782:MTT formazan; D003300:Copper; D011190:Potassium Cyanide; C000520:cuprous oxide