Abstract: Background – Transmucosal glucagon has potential use in at-home and in-hospital emergencies to treat life-threatening hypoglycemia in cats. Objectives – To evaluate the effect of transmucosal glucagon (Baqsimi™) on blood glucose concentrations (BG) in healthy cats and describe adverse reactions to its administration. Animals – Six healthy, purpose-bred cats.Methods – Randomized, controlled, crossover study. Transmucosal glucagon was administered intranasally and rectally and compared to intranasal placebo. Blood was collected at -15 and -1 minutes before, and 5, 15, 25, 35, 45 and 60 minutes after glucagon administration for evaluation of BG, plasma glucagon (pGlucagon), and plasma potassium (K+) concentrations. Stress scores and adverse effects were recorded at all time points. Results – pGlucagon in the nasal and rectal groups increased from baseline (median [range]) (12.2 [3.5-44.1] mmol/L nasal and 6.9 [2.9-21.1] mmol/L rectal), to 218.5 [7.9-349.8] mmol/L (P=0.02) and 349.8 [67.4-349.8] mmol/L (P=0.01) respectively, 15 minutes after administration. BG increased from baseline (101 [91-110] mg/dL) 15 minutes after nasal (137.5 [104-251] mg/dL, P=0.006) and rectal (229 [99-285] mg/dL P=0.002) administration. K+ decreased from baseline (3.8 [3.6-4.1] mmol/L nasal and 3.7 [3.5-3.9] mmol/L rectal) to 3.4 [3.1-3.6] mmol/L (P=0.04) at 15 minutes with nasal administration, and with rectal administration, to 3.2 [3.1-3.6] mmol/L (P=0.04) at 15 minutes and 3.1 [2.9-3.4] mmol/L (P=0.01) at 25 minutes. No significant changes were detected in the placebo group. No serious adverse effects were noted.Conclusions and clinical importance – Transmucosal glucagon is effective in raising BG with minimal side effects in healthy cats.