Microelectrode array recordings of cardiac action potentials as a high throughput method to evaluate pesticide toxicity.
The threat of environmental pollution, biological warfare agent dissemination and new diseases in recent decades has increased research into cell-based biosensors. The creation of this class of sensors could specifically aid the detection of toxic chemicals and their effects in the environment, such as pyrethroid pesticides. Pyrethroids are synthetic pesticides that have been used increasingly over the last decade to replace other pesticides like DDT. In this study we used a high-throughput method to detect pyrethroids by using multielectrode extracellular recordings from cardiac cells. The data from this cell-electrode hybrid system was compared to published results obtained with patch-clamp electrophysiology and also used as an alternative method to further understand pyrethroid effects. Our biosensor consisted of a confluent monolayer of cardiac myocytes cultured on microelectrode arrays (MEA) composed of 60 substrate-integrated electrodes. Spontaneous activity of these beating cells produced extracellular field potentials in the range of 100 microV to nearly 1200 microV with a beating frequency of 0.5-4 Hz. All of the tested pyrethroids; alpha-Cypermethrin, Tetramethrin and Tefluthrin, produced similar changes in the electrophysiological properties of the cardiac myocytes, namely reduced beating frequency and amplitude. The sensitivity of our toxin detection method was comparable to earlier patch-clamp studies, which indicates that, in specific applications, high-throughput extracellular methods can replace single-cell studies. Moreover, the similar effect of all three pyrethroids on the measured parameters suggests, that not only detection of the toxins but, their classification might also be possible with this method. Overall our results support the idea that whole cell biosensors might be viable alternatives when compared to current toxin detection methods.
Natarajan
A
A
Nanoscience Technology Center, University of Central Florida, Orlando, FL 32826, USA.
Molnar
P
P
Sieverdes
K
K
Jamshidi
A
A
Hickman
J J
JJ
D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D015374:Biosensing Techniques; D002642:Chick Embryo; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation* / Q000379:methods; D004594:Electrophysiology; D006321:Heart / Q000187:drug effects*; D007306:Insecticides / Q000633:toxicity; D008839:Microelectrodes; D032342:Muscle Cells / Q000187:drug effects / Q000502:physiology; D010575:Pesticides / Q000633:toxicity*; D011722:Pyrethrins / Q000633:toxicity; D013499:Surface Properties
D007306:Insecticides; D010575:Pesticides; D011722:Pyrethrins