The major limiting factor in the successful management of type 1 diabetes is Recurrent Hypoglycaemia (RH), resulting from supraphysiological insulin supplementation, which attenuates the Counterregulatory Response (CRR) to future hypoglycaemic episodes. It is now widely accepted that the hypothalamus contributes to hypoglycaemia detection and activation of the CRR (1). Within the hypothalamus, glucose-sensing (GS) neurons exist which are thought to contain the same GS cellular machinery as pancreatic β-cells, such as ATP-sensitive K+ channels (KATP), glucokinase, glucose transporter proteins and AMPK (2). We hypothesise that disruption of normal function in these specialised neurons may occur following RH. We have previously shown that mouse hypothalamic GT1-7 cells are GS neurons (3) and display impaired electrical activity to low glucose following RH in association with impaired AMPK activation (4). Here we have investigated adaptations in glucose metabolism and KATP channel activity as possible mediators of altered electrical activity following RH. GT1-7 cells were exposed to 3 hours of low (0.1 mM) glucose for 3 days to model RH, with cells being assayed on day 4. Cellular Oxygen Consumption Rate (OCR) and Extracellular Acidification Rate (ECAR) were measured under euglycaemic conditions using an XF24 extracellular flux analyser (Seahorse Bioscience, Inc.). The ATP/ADP ratio within cells was examined using bioluminescence and single KATP channel activity was monitored through the use of inside-out patch-clamp electrophysiology. All data sets are expressed as mean ± standard error of the mean. Statistical analysis was performed using either Student’s t-test or ANOVA. Following RH, GT1-7 cells displayed significantly reduced basal OCR (RH = 4.60 ± 0.51 vs. Cont = 7.97 ± 0.63 pmoles O2/min/mg; p < 0.001; n=8) and ECAR (RH = 0.59 ± 0.07 vs. Cont = 1.03 ± 0.14 mpH/min/mg; p < 0.05; n=5). The ATP/ADP ratio fell significantly from 2.5 to 0.1 mM glucose in control cells (2.34 ± 0.14; 1.80 ± 0.17, respectively; p < 0.05; n=7). However, the basal ATP/ADP ratio was found to be lower following RH and did not fall further in response to 0.1 mM glucose, applied acutely (1.91 ± 0.18; 1.78 ± 0.27, respectively; n=7). The adaptations in the energy status of GT1-7 cells following RH also coincided with a significant reduction in the open channel probability of the KATP channel (N.Po) in response to MgADP (RH = 0.003 ± 0.001 vs. Cont = 0.072 ± 0.010; p < 0.05; n=5). We conclude that RH produces a hypometabolic state, whereby the KATP channel should be active but remains closed. Failure to reduce the ATP/ADP ratio during acute low glucose exposure in association with an attenuated sensitivity of the KATP channel to activation by MgADP may contribute to defective GS of hypothalamic neurons following RH.