C57bl 6

Six-week-old male mice (C57BL/6) were purchased from Koatech (Pyeongtaek, South Korea). Approximately 1.85 MBq of ⁶⁴Cu-labeled liposomes with clodronate (500 µg, 1.39 µmol) were injected through tail vein into seven-week-old normal mice (C57BL/6) anesthetized with 2% isoflurane, to confirm the in vivo biodistribution. The number of L and ML to be injected was determined based on the number of CL and CML injected. The PET scan images were acquired at different time points (0, 2, 8, and 24 h) after injection, using a preclinical PET/X-ray scanner (GENISYS4, Sofie Bioscience, California, USA). PET imaging was conducted using the InVivoScope software (version 2.0). The region of interest was calculated using the AMIDE software to quantitatively evaluate the uptake in the blood pool and liver. The time-activity curve was fitted based on % ID/g at each time point.

We obtained male 5xFAD transgenic mice [B6.Cg-Tg (APPSwFlLon, PS1*M146L*L286V) 6799Vas/Mmjax; JAX MMRRC stock number: 34848] from The Jackson Laboratory (Bar Harbor, ME, USA). Aβ₄₂ production was identified in the entire brain at 2 months. Wild-type male mice (C57BL/6) were purchased from Koatech (Pyeongtaek, South Korea). For pineal gland harvesting, the mice with 5 months of age were sacrificed under ether anesthesia. We took samples from all mice at the same time of the day [Zeitgeber time (ZT) 0.5] under conditions of 12 h in light and 12 h in dark. There was no noticeable difference in the phenotype between the pineal glands of wild type and 5xFAD mice. The pineal glands from five to seven mice were pooled in each sample, and four samples in each of wild type and 5xFAD group were prepared for the analysis. The experiment was performed in accordance with the recommendations of “96 Guidance for Animal Experiments” established by the Animal Ethics Committee at Chonnam National University. The experimental protocols were approved by the Animal Ethics Committee at Chonnam National University.

All experimental protocols were approved by the Institutional Animal Care and Use Committees of DGIST (DGIST-IACUC_0104). All applicable guidelines for the care and use of laboratory animals from the National Institutes of Health Guide were followed. Only male animals were used in this study. Adult mice (C57/BL6, 2-month-old) were obtained from KOATECH (Daegu, Korea). 5xFAD transgenic mice harboring the mutated human APP (695 amino acids) and human PS1 genes (Tg6799, 3–4 months old) were obtained from Prof. K.A. Chang (Gachon Medical School, Incheon, Korea). All behavioral, physiological, and histological analyses using animals in this study were blind tested.

C57BL/6 8-week-old female mice were purchased from KOATECH (Pyeongtaek, Korea) and randomly divided into two groups (n = 10 per group). All animals were housed in a temperature-controlled room (22°C) under artificial illumination with a 12-h light/dark cycle and 55% relative humidity. The mice were provided access to food and water ad libitum. All operational procedures were performed under deep anesthesia. OVX was performed in one group (OVX group) of 8-week-old mice. Incisions and sutures on the back of mice were performed in the other group (Sham group).