Date Received: 03-04-2026 / Date Accepted: 22-09-2026 / Date Published: 26-10-2026
Selecting appropriate reference genes is crucial for ensuring the reliability of RT-qPCR-based gene expression analysis. This study aimed to compare β-actin and GAPDH as reference genes for evaluating the expression of BDNF and BACE1 in the mouse hypothalamus under a fasting-refeeding model. Healthy male mice were divided into three groups: control, 48-hour complete fasting (with free access to water), and 4-hour refeeding after 48 hours of fasting. Hypothalamic samples were collected, total RNA was extracted, cDNA was synthesized, and gene expression was analyzed by RT-qPCR. When normalized to -actin, BDNF expression was significantly decreased in the 48-hour fasting group and was no longer significantly different from the control group after 4 hours of refeeding. In contrast, when normalized to GAPDH, BDNF expression showed no significant changes in either treatment group. For BACE1, expression levels did not differ significantly among groups and yielded consistent results regardless of normalization with either reference gene. These findings indicate that -actin and GAPDH are not fully equivalent as reference genes in the fasting–refeeding model, and β-actin may be more suitable for evaluating genes sensitive to metabolic status in the mouse hypothalamus.
