Dec 17, 2015 - Abstract. Background/Aims: Kir2.1 (KCNJ2) channels are expressed in neurons, skeletal muscle and cardiac tissue and maintain the resting membrane potential. The activity of those channels is regulated by diverse signalling molecules. The present study explored whether Kir2.1 channels are sensitive to.

1. Serial Argile 2.1 Download

Co-expression of wild-type SPAK increased the inwardly rectifying K + current in Kir2.1-expressing Xenopus oocytes. A: Representative original tracings showing currents in Xenopus oocytes injected with water (a), expressing wild-type SPAK alone (b), expressing Kir2.1 alone (c) or expressing Kir2.1 with additional co-expression of wild-type SPAK (d). The voltage protocol is shown (not to scale). B: Arithmetic means ± SEM (n = 9-37) of the current (I) as a function of the potential difference across the cell membrane (V) in Xenopus oocytes injected with water (white circles), expressing SPAK alone (black triangles), expressing Kir2.1 alone (white squares) or expressing Kir2.1 with SPAK (black squares). C, D: Arithmetic means ± SEM (n = 9-37) of (C) maximal current and (D) conductance calculated by linear fit of I/V-curves shown in B between -150 mV and -120 mV in Xenopus oocytes injected with water (dotted bar), expressing SPAK alone (grey bar) or expressing Kir2.1 without (white bar) or with (black bar) additional co-expression of wild-type SPAK. The effect of wild-type SPAK on Kir2.1 was mimicked by constitutively active T233ESPAK but not by WNK insensitive T233ASPAK or catalytically inactive D212ASPAK.

A: Representative original tracings showing currents in Xenopus oocytes injected with water (a) expressing Kir2.1 alone (b), expressing Kir2.1 together with constitutively active T233ESPAK (c), with WNK insensitive T233ASPAK (d) or with catalytically inactive D212ASPAK (e). B: Arithmetic means ± SEM (n = 13-33) of the current (I) as a function of the potential difference across the cell membrane (V) in Xenopus oocytes injected with water (white circles), expressing Kir2.1 alone (white squares) or expressing Kir2.1 together with constitutively active T233ESPAK (black squares), inactive T233ASPAK (grey diamonds) or catalytically inactive D212ASPAK (black triangles). C, D: Arithmetic means ± SEM (n = 13-33) of (C) maximal current and (D) conductance calculated by linear fit of I/V-curves shown in B between -150 mV and -120 mV in Xenopus oocytes injected with water (dotted bar), expressing Kir2.1 alone (white bar), expressing Kir2.1 with constitutively active T233ESPAK (black bar), or expressing Kir2.1 with inactive T233ASPAK (light grey bar), or D212ASPAK (dark grey bar). Co-expression of SPAK increased the Kir2.1 protein abundance within the plasma membrane of Xenopus oocytes.

Serial Argile 2.1 Download

Arithmetic means ± SEM (n = 84-88) from the normalized data of Kir2.1-HA protein abundance as determined by chemiluminescence in the plasma membrane of Xenopus oocytes injected with water (dotted bar) or expressing Kir2.1-HA without (white bar) or with (black bar) additional expression of wild-type SPAK. Effect of brefeldin A on inwardly rectifying K + current in Kir2.1 and SPAK expressing oocytes. A: Representative original tracings showing currents in Xenopus oocytes expressing Kir2.1 alone (a,b) or expressing Kir2.1 together with-wild type SPAK (c,d), prior to (a,c) and following (b,d) a 24 hours incubation with 5 µM brefeldin A. B: Arithmetic means ± SEM (n = 14-21) of the current (I) as a function of the potential difference across the cell membrane (V) in Xenopus laevis oocytes expressing Kir2.1 alone (white square) or expressing Kir2.1 together with OSR1 (black squares), prior to (white circles) and following (black circles) a 24 hours incubation with 5 µM brefeldin A. C, D: Arithmetic means ± SEM (n = 14-21) of (C) maximal current and (D) conductance calculated by linear fit of respective I/V-curves between -150 mV and -120 mV in Xenopus oocytes expressing Kir2.1 alone (white bars) or expressing Kir2.1 together with wild type SPAK (black bars) prior to (left bars, 0h) and following a 24 hours incubation with 5 µM brefeldin A (right bars). Co-expression of wild-type OSR1 increased the inwardly rectifying K + current in Kir2.1-expressing Xenopus oocytes. A: Representative original tracings showing currents in Xenopus oocytes injected with water (a), expressing wild-type OSR1 alone (b), expressing Kir2.1 alone (c) or expressing Kir2.1 with additional co-expression of wild-type OSR1 (d).

The voltage protocol is shown (not to scale). B: Arithmetic means ± SEM (n = 9-35) of the current (I) as a function of the potential difference across the cell membrane (V) in Xenopus oocytes injected with water (white circles), expressing OSR1 alone (black triangle), expressing Kir2.1 alone (white squares) or expressing Kir2.1 with OSR1 (black squares). C, D: Arithmetic means ± SEM (n = 9-35) of (C) maximal current and (D) conductance calculated by linear fit of I/V-curves shown in B between -150 mV and -120 mV in Xenopus oocytes injected with water (dotted bar), expressing OSR1 alone (grey bar) or expressing Kir2.1 without (white bar) or with (black bar) additional co-expression of wild-type OSR1. The effect of wild type OSR1 on Kir2.1 was mimicked by constitutively active T185EOSR1 but not by WNK insensitive T185AOSR1 or catalytically inactive D164AOSR1.

A: Representative original tracings showing currents in Xenopus oocytes injected with water (a) expressing Kir2.1 alone (b), expressing Kir2.1 together with constitutively active T185EOSR1 (c), with WNK insensitive T185AOSR1 (d) or with catalytically inactive D164AOSR1 (e). B: Arithmetic means ± SEM (n = 9-35) of the current (I) as a function of the potential difference across the cell membrane (V) in Xenopus laevis oocytes injected with water (white circles), expressing Kir2.1 alone (white squares) or expressing Kir2.1 together with constitutively active T185EOSR1 (black squares), inactive T185AOSR1 (grey diamonds) or catalytically inactive D164AOSR1 (black triangle). C, D: Arithmetic means ± SEM (n = 17-35) of (C) maximal current and (D) conductance calculated by linear fit of I/V-curves shown in B between -150 mV and -120 mV in Xenopus oocytes injected with water (dotted bar), expressing Kir2.1 alone (white bar), expressing Kir2.1 with constitutively active T185EOSR1 (black bar), or expressing Kir2.1 with inactive T185AOSR1 (light grey bar), or D164AOSR1 (dark grey bar). Effect of brefeldin A on inwardly rectifying K + current in Kir2.1 and OSR1 expressing oocytes. A: Representative original tracings showing currents in Xenopus oocytes expressing Kir2.1 alone (a,b) or expressing Kir2.1 together with wild type OSR1 (c,d), prior to (a,c) and following (b,d) a 24 hours incubation with 5 µM brefeldin A. B: Arithmetic means ± SEM (n = 14-20) of the current (I) as a function of the potential difference across the cell membrane (V) in Xenopus laevis oocytes expressing Kir2.1 alone (white squares) or expressing Kir2.1 together with OSR1 (black squares), prior to (white circles) and following (black circles) a 24 hours incubation with 5 µM brefeldin A. C, D: Arithmetic means ± SEM (n = 14-20) of (C) maximal current and (D) conductance calculated by linear fit of respective I/V-curves between -150 mV and -120 mV in Xenopus oocytes expressing Kir2.1 alone (white bars) or expressing Kir2.1 together with wild type OSR1 (black bars) prior to (left bars, 0h) and following a 24 hours incubation with 5 µM brefeldin A (right bars).

Accepted: November 28, 2015 Published online: December 17, 2015 Issue release date: December 2015 Number of Print Pages: 14 Number of Figures: 7 Number of Tables: 0 ISSN: (Print) eISSN: (Online) For additional information: Open Access License / Drug Dosage / Disclaimer This article is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND). Usage and distribution for commercial purposes as well as any distribution of modified material requires written permission. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.

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