The use of microelectrode array (MEA) to study rat peritoneal mast cell activation.
We performed this study to demonstrate the applicability of the microelectrode array (MEA) to study electrophysiological changes of rat peritoneal mast cells in the presence of compound 48/80 under normal, Ca(2+)-free, Ca(2+)-free with EDTA, and Cl(-)-free conditions. The use of high extracellular K(+) (KCl, 150 mM), charybdotoxin (ChTX, 100 nM), and Cl(-)-free containing ChTX buffers verified that the hyperpolarizing signal was due to the activation of mainly K(+) and, to a lesser extent, Cl(-) channels. Compound 48/80 concentration-dependently shortened the latent periods (the onset of response) and increased both the spatial (the K(+) and Cl(-) hyperpolarizing field potentials, HFP) and temporal measurements (the duration of response). Ca(2+)-free buffer had no effect on the latent period of compound 48/80 but increased the HFP at high concentrations. The latent period increased while the HFP diminished when cells were equilibrated in Ca(2+)-free buffer containing EDTA. Durations of the HFP were generally longer when cells were in either Ca(2+)-free or Ca(2+)-free containing EDTA buffers than when cells were in normal buffer. The EC(50) values confirmed that effects were only affected in Ca(2+)-free buffer containing EDTA but not in Ca(2+)-free or Cl(-)-free buffers, further reinforcing the hypothesis that the presence of Ca(2+) is not essential to the action of compound 48/80. The present study is the first application of MEA to study rat peritoneal mast cells, and our results indicate that it could be of value in future pharmacological research on other non-excitable cells.
Yeung
Chi-Kong
CK
Department of Pharmacology, Faculty of Medicine, The Chinese University of Hong Kong (CUHK), Shatin, Hong Kong. chi-kong_yeung@lycos.com
Law
Jessica Ka-Yan
JK
Sam
Sze-Wing
SW
Ingebrandt
Sven
S
Lau
Hang-Yung Alaster
HY
Rudd
John Anthony
JA
Chan
Mansun
M
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D018118:Chloride Channels / Q000502:physiology; D004492:Edetic Acid / Q000494:pharmacology; D008297:Male; D008407:Mast Cells / Q000502:physiology*; D046228:Microarray Analysis / Q000379:methods*; D008839:Microelectrodes; D010529:Peritoneal Cavity / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D003189:p-Methoxy-N-methylphenethylamine / Q000494:pharmacology
D018118:Chloride Channels; D003189:p-Methoxy-N-methylphenethylamine; D004492:Edetic Acid; D002118:Calcium