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KMID : 0358619950290020301
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Korean Journal of Physiology
1995 Volume.29 No. 2 p.301 ~ p.307
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Electrophysiological Responses of Receptor Expressed on HEK293 Cells
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Kim Jin-Hyuk
Koh Young-Ik
Chin He-Min
Lee Yong-Sung
Cho Yeul-Hee
Kim Kee-Soon
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Abstract
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To explore electrophysiological properties of the receptors artificially expressed in the mammalian cell, effect of an opioid agonist DPDPE on the voltage-sensitive outward currents was examined in the HEK293 (human embryonic kidney) cells transfected with receptor cDNA cloned from NG-108-15 cDNA library. Also studied were effects of 8-bromo-cyclic AMP and naloxone on DPDPE-induced changes in the voltage sensitive outward current. The voltage sensitive outward currents were recorded using perforated patch technique at room temperature. In the non-transformed HEK293 cells, DPDPE did not alter voltage sensitive outward current, indicating that no native receptor had been developed. However, DPDPE remarkably increased the voltage sensitive outward current in the transformed HEK293 cells. The increment in voltage sensitive outward current peaked in after DPDPE application, and the maximum DPDPE-activated outward current was recorded when the membrane potential was depolarized to +70mv. Following pretreatment of the transformed HEK293 cells with 1 mM 8-bromo-cyclic AMP, DPDPE failed to increase the voltage sensitive outward currents. On the other hand, naloxone completely abolished DPDPE-activated voltage sensitive outward current in the transformed HEK293 cells. The results of present study suggest that in the transformed HEK293 cells an activation of the receptors by an opioid agonist DPDPE increases the voltage-sensitive potassium current as a result of decrement in cyclic AMP level.
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KEYWORD
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receptor, HEK293 cell, NG108-15 cell
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