It is known that autonomic nerve activity (ANA) affects glucose metabolism by regulating the secretion of insulin and glucagon. Sympathetic nerve stimulation results in increased blood glucose levels. ANA also showed a circadian variation, and sympathetic nerve activity was minimal at night and began to rise at arousal. Therefore, a drastic alteration in ANA around wake-up would be associated with glycemic variability (GV) known risk factor for cardiovascular disease. We investigated the relation between ANA around wake-up and either morning or daily GV. Methods: We simultaneously performed Holter ECG and continuous glucose monitoring system in 41 patients with type 2 diabetes (T2D). ANA was assessed by heart rate variability (HRV) analysis. Delta (D) wake-up was defined as the difference between the maximum and minimum value during 1 h before and after wake-up time, before breakfast. Results: D of low frequency/high frequency (LF/HF) around wake-up time (D LF/HF wake-up) was positively associated with D glucose wake-up , standard deviation (SD) glucose wake-up , the mean amplitude of glucose excursions (MAGE 24h), and SD glucose 24h after adjustment for age, sex, BMI, the duration of diabetes, and the prevalence of diabetic polyneuropathy (b = 0.47, p = 0.011, b = 0.48, p = 0.009, b = 0.54, p = 0.002 and b = 0.41, p = 0.0025, respectively). No association was found between D LF/HF wake-up and either mean blood glucose for 24 h, or HbA1c as parameters of chronic hyperglycemia. Conclusions: In T2D, the fluctuation in fasting sympathetic nerve activity around wake-up was positively associated with not only morning but also daily GV.