Temperature: an important experimental variable in studying PKC modulation of ligand-gated ion channels

Brain Research
Tina K MachuN J Leidenheimer

Abstract

Amphibian oocyte and mammalian heterologous expression systems are often used to investigate the function of recombinant ion channels using electrophysiological techniques. Although both systems have yielded important information, the results obtained in these systems are sometimes conflicting. Oocytes and mammalian cells differ in their physiological temperature requirements. While room temperature is within the physiological temperature range for oocytes, this temperature is far below that required by mammalian cells. Since electrophysiological studies are often performed in both oocytes and mammalian cells at room temperature, we sought to determine if recording temperature could be a factor in some disparate results obtained in these cell types. For these studies, we examined phorbol ester modulation of GABA(A) and glycine receptors. Consistent with the literature, at room temperature, PMA (phorbol 12-myristate 13-acetate) produced a large reproducible decrease in the peak amplitude of GABA and glycine-gated currents in Xenopus oocytes. In contrast, PMA was ineffective in modulating these heterologously expressed receptors at room temperature in human embryonic kidney (HEK) 293 cells. However, when electrophysiological expe...Continue Reading

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Citations

Sep 25, 2007·Biochemistry·Renqi HuangNancy J Leidenheimer
Oct 10, 2013·The European Journal of Neuroscience·Damian P Bright, Trevor G Smart
Jun 6, 2018·Frontiers in Molecular Neuroscience·Ulrike BreitingerHans-Georg Breitinger
Dec 7, 2007·Analytical Chemistry·Maria MillingenOwe Orwar

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