Transcriptional profiling of CD11b+ microglia isolated from cerebellum and olfactory bulb-removed brains of SAMP8 and SAMR1 mice at 2, 5, and 9 months of age.

SAMP8 mice serve as an accelerated aging model characterized by inflammation and neurological memory deficits, in contrast to their control SAMR1 counterparts. To investigate the role of microglia in aging, we conducted a comprehensive analysis of microglial heterogeneity using CITE-seq. CD11b positive cells were extracted from cerebellum and olfactory bulb-removed brains at key time points: 2 months, 5 months, and 9 months of age. Overall design: At 2, 5, and 9 months of age, brains of SAMP8 and SAMR1 mice were prepared by removing the cerebellum and olfactory bulbs. The remaining brain tissue was then physically and enzymatically dissociated in presence of Actinomycin D and Brefeldin A. Following dissociation, CITE-seq antibodies (ADTs and HTOs) and CD11b-PE/Cy5 staining were used. The DAPI-CD11b+ cells were subsequently isolated using fluorescence-activated cell sorting (FACS).