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. 2017 Apr 4;7:640. doi: 10.1038/s41598-017-00699-3

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Kir6.2/SUR1 combination of K_ATP channels in the SCN neurones. (A) RT-PCR analysis indicating the expression of mRNA for pore-forming Kir6.2 and sulfonylurea subunits 1 in the SCN. Positive controls were performed using cDNA from rat brain. The expected PCR product sizes for Kir6.1, Kir6.2, SUR1, and SUR2 were 411, 385, 388, and 501 bp, respectively. Negative controls were performed using RT products with omission of reverse transcriptase (RT-) to examine the contamination of genomic DNA. (B) Cell-attached recordings showing the effects of the sulfonylurea on spontaneous firing of the SCN neurones (ZT 4–15). Firing responses of a representative cell to 200 μM tolbutamide (top left panel), 200 μM diazoxide (top middle panel), and 0.1 μM glibenclamide (top right panel). Note the lack of recovery of spontaneous firing after washout of glibenclamide. Daytime recordings (ZT 7). Bottom left panel: summary of experiments showing a moderate increase in firing rate by tolbutamide. Baseline spontaneous firing rate: 2.7 ± 0.3 Hz (n = 50). Bottom right panel: summary of experiments showing a decrease in firing rate by diazoxide. Baseline spontaneous firing rate: 3.0 ± 0.3 Hz (n = 17). *P < 0.05, ***P < 0.001.