Dantrolene treatment groups

Age-matched male and female mice were randomly divided into experimental groups when they were genotyped around 1 month of age (Fig. 1). The Early Treatment Groups (ETG), including intranasal dantrolene (IN-DAN), subcutaneous dantrolene (SQ-DAN), intranasal vehicle (IN-VEH), and no treatment (CON, negative control), were treated once a day, 3×/week (Monday, Wednesday, and Friday) beginning at 2 months of age, before the onset of intracellular amyloid pathology and any cognitive dysfunction [20]. The Late Treatment Groups (LTG), intranasal dantrolene (IN-DAN) and subcutaneous dantrolene (SQ-DAN), began the same treatment at 6 months of age, well after the onset of extracellular amyloid plaques accumulation and cognitive dysfunction [20]. The vehicle was made fresh and contained all inactive ingredients in Ryanodex. Fresh dantrolene was made every time before administration with the vehicle for intranasal (5 mg/ml) and subcutaneous (1 mg/ml) administration. All mice continued to receive treatment until they were euthanized at 12 months of age.

Experimental design. Timeline for treatments, behavioral tests, and euthanasia. Twelve experimental groups were designed based on genotype (5XFAD, WT), age when treatment began (Early Treatment (ETG), Late Treatment (LTG) groups), and the administration route of dantrolene (Intranasal, Subcutaneous).