The compound action potential (CAP) recorded from ex vivo mouse optic nerve is a popular model used to study nervous system metabolism. A recent advance in this technique has been recording from both optic nerves simultaneously using paired sets of stimulating and recording electrodes to acquire CAPs from both nerves simultaneously. The method has advantages in studies where one nerve can act as the control for its treated partner. We report on recordings from pairs of sciatic nerve from which we derived a mathematically based rationale that demonstrates decreased experimental variability whilst maintaining statistical power and reducing animal use. All procedures were carried out in accordance with the Animals (Scientific Procedures) Act 1986 under appropriate authority of project and personal licenses. Adult male CD-1 mice were killed by cervical dislocation and decapitated. Sciatic nerves were dissected, placed in a superfusion chamber, with aerated aCSF containing 10 mM glucose. The stimulus evoked A fibre CAP was evoked with supra-maximal stimuli. We employed two separate recording set ups, a conventional one in which single sciatic nerve CAPs were recorded, and a set up where pairs of sciatic nerve were recorded simultaneously. The protocol involved stimulating nerves continuously at a rate of 1 Hz during the baseline period, followed by a period of aglycaemia, during which nerves were stimulated at either 1 Hz or 50 Hz. Increasing the stimulus frequency to 50 Hz accelerated A fibre CAP failure during aglycaemia, which was assessed as the latency for the CAP to fall to 95% of its baseline value. In recordings from single nerves stimulated at 1 Hz, the mean latency to CAP failure was 99.4 ± 28.0 minutes (n = 12) versus 79.7 ± 24.9 minutes (n = 12) for nerves stimulated at 50 Hz, a significant difference (p = 0.040: two sample unequal variance t-test). In recordings from pairs of nerves the equivalent values were 96.4 ± 17.2 minutes (n = 12) versus 80.1 ± 18.9 minutes (n = 12), p = 0.019. The relative standard deviation (RSD), which is the coefficient of variation expressed as a percentage showed a statistically significant decrease from pairs of nerves compared to single nerve recordings (p = 0.015, n = 12, unpaired t-test). Recordings from pairs of nerves simultaneously has allowed us to reduce the number of mice used in experiments designed to test the role of glycogen in supporting axonal conduction during periods of aglycaemia in sciatic nerve. Mathematical analysis of our data showed that for equivalent number of nerves, when recordings derived from paired recordings the p value of comparisons was lower and power was higher meaning that under these conditions the risk of making Type I and Type II error was reduced compared to recordings from single nerves.