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1- Department of Physiology, School of Medicine, Arak University of Medical Sciences, Arak, Iran.
2- Neurophysiology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
3- Department of Biochemistry, Pasteur institute of Iran, Tehran, Iran.
4- Cellular and Molecular Research Center and Department of Physiology, School of Medicine, Iran University of Medical Sciences, Tehran Iran.
5- Electrophysiology Research Center, Neuroscience Institute, Tehran University of Medical sciences, Tehran, Iran.
2- Neurophysiology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
3- Department of Biochemistry, Pasteur institute of Iran, Tehran, Iran.
4- Cellular and Molecular Research Center and Department of Physiology, School of Medicine, Iran University of Medical Sciences, Tehran Iran.
5- Electrophysiology Research Center, Neuroscience Institute, Tehran University of Medical sciences, Tehran, Iran.
Abstract:
Several types of ion channels found in the plasma membrane have also been identified in the membranes of intracellular organelles. These ion channels, including potassium channels, play a crucial role in regulating intracellular ion homeostasis. An ATP-sensitive potassium channel (KATP) with various functional roles has been identified in the endo/sarcoplasmic reticulum membranes of both excitable and non-excitable cells. Our Previous studies have investigated the electro pharmacological and molecular properties of KATP and Bkca+2 channels in the rough endoplasmic reticulum (RER) of rat hepatocytes. In this study, for the first time, we described the electro-pharmacological and molecular properties of the RER ATP-sensitive potassium channel in rat brain cells using an incorporated single channel in the planar lipid bilayer and western blotting analysis. The results of the study revealed the presence of an ATP-sensitive potassium channel with a conductance of 306 pS, and the open probability was found to be voltage-independent at a holding potential ranging from +40 to -60 in an asymmetric solution (200/50 mM KCl; cis/trans). Additionally, we observed that adding ATP (2.5 mM) to the positive and negative potentials and 100 μM glibenclamide to the positive voltages inhibited the channel activity. Interestingly, the addition of 100 mM 5-HD and 100 nM charybdotoxin to the cis side did not affect the channel behavior. Furthermore, a western blot analysis provided evidence of the expression of Kir6.2, Kir6.1, SUR1, and/or SUR2B, but not SUR2A, in the RER of rat brain fractions. In this study, we provide strong evidence for the existence of an ATP-sensitive potassium channel on the RER membrane of rat brain cells, displaying different pharmacological properties than those classically described for the plasma membrane and other intracellular organelles.
Keywords: Potassium channels, RER KATP channel, Bilayer lipid membrane, Single channel recording
Type of Study: Original |
Subject:
Cellular and molecular Neuroscience
Received: 2023/07/15 | Accepted: 2024/04/13
Received: 2023/07/15 | Accepted: 2024/04/13
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