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Workflow and cluster identification. Ten-week-old male Sprague Dawley (SD) rats were obtained from Charles River, of which 5 were randomly assigned to the sedentary group and 5 were assigned to the exercise donor groups. Exercise donor rats underwent an exercise regimen 5 days a week for 1 month (A) . After the last bout of exercise, rats were exsanguinated and plasma exosomes isolated (B) . 11-month-old male rats (retired breeders) were fitted unilaterally on the left anatomical side of the calvarium with an <t>intracerebroventricular</t> <t>(ICV)</t> cannula by Charles River and infused daily for 7 days with 48 μg of exercise or sedentary donor plasma exosomes in 10 μL of PBS under light anesthesia. After the last infusion, the rats were euthanized and perfused with ice-cold saline. Brains were extracted, and a region enriched for basal ganglia was dissected (C) . Tissue was rapidly frozen on dry ice. For single-nucleus RNA-seq, the tissue was pulverized under liquid nitrogen using a mortar and pestle kept on dry ice. The samples were then processed to isolate nuclei. Isolated nuclei were validated with flow cytometry and sequenced (D). A total of 26 distinct cell clusters were identified, including cells of primary interest with their cell counts per cluster: Astrocytes 16,079, dopaminergic neurons from substantia nigra pars compacta 1,591, Gad2 neurons 8,284, medium spiny neurons 2,343, microglia 12,529, and oligodendrocytes 45,600 (E) . Workflow diagram was created with BioRender .
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Workflow and cluster identification. Ten-week-old male Sprague Dawley (SD) rats were obtained from Charles River, of which 5 were randomly assigned to the sedentary group and 5 were assigned to the exercise donor groups. Exercise donor rats underwent an exercise regimen 5 days a week for 1 month (A) . After the last bout of exercise, rats were exsanguinated and plasma exosomes isolated (B) . 11-month-old male rats (retired breeders) were fitted unilaterally on the left anatomical side of the calvarium with an intracerebroventricular (ICV) cannula by Charles River and infused daily for 7 days with 48 μg of exercise or sedentary donor plasma exosomes in 10 μL of PBS under light anesthesia. After the last infusion, the rats were euthanized and perfused with ice-cold saline. Brains were extracted, and a region enriched for basal ganglia was dissected (C) . Tissue was rapidly frozen on dry ice. For single-nucleus RNA-seq, the tissue was pulverized under liquid nitrogen using a mortar and pestle kept on dry ice. The samples were then processed to isolate nuclei. Isolated nuclei were validated with flow cytometry and sequenced (D). A total of 26 distinct cell clusters were identified, including cells of primary interest with their cell counts per cluster: Astrocytes 16,079, dopaminergic neurons from substantia nigra pars compacta 1,591, Gad2 neurons 8,284, medium spiny neurons 2,343, microglia 12,529, and oligodendrocytes 45,600 (E) . Workflow diagram was created with BioRender .

Journal: Frontiers in Aging Neuroscience

Article Title: Intracerebroventricular infusion of exercise donor plasma exosomes induces molecular changes indicating exercise-like adaptations in basal ganglia cells of older male rats

doi: 10.3389/fnagi.2026.1766698

Figure Lengend Snippet: Workflow and cluster identification. Ten-week-old male Sprague Dawley (SD) rats were obtained from Charles River, of which 5 were randomly assigned to the sedentary group and 5 were assigned to the exercise donor groups. Exercise donor rats underwent an exercise regimen 5 days a week for 1 month (A) . After the last bout of exercise, rats were exsanguinated and plasma exosomes isolated (B) . 11-month-old male rats (retired breeders) were fitted unilaterally on the left anatomical side of the calvarium with an intracerebroventricular (ICV) cannula by Charles River and infused daily for 7 days with 48 μg of exercise or sedentary donor plasma exosomes in 10 μL of PBS under light anesthesia. After the last infusion, the rats were euthanized and perfused with ice-cold saline. Brains were extracted, and a region enriched for basal ganglia was dissected (C) . Tissue was rapidly frozen on dry ice. For single-nucleus RNA-seq, the tissue was pulverized under liquid nitrogen using a mortar and pestle kept on dry ice. The samples were then processed to isolate nuclei. Isolated nuclei were validated with flow cytometry and sequenced (D). A total of 26 distinct cell clusters were identified, including cells of primary interest with their cell counts per cluster: Astrocytes 16,079, dopaminergic neurons from substantia nigra pars compacta 1,591, Gad2 neurons 8,284, medium spiny neurons 2,343, microglia 12,529, and oligodendrocytes 45,600 (E) . Workflow diagram was created with BioRender .

Article Snippet: A total of 10 retired male Sprague Dawley (SD) breeders (11 months of age) were obtained with pre-implanted intracerebroventricular (ICV) cannulas from Charles River Laboratories (Wilmington, MA) and handled in accordance with the VA New Jersey Health Care System Institutional Animal Care and Use Committee (IACUC) guidelines.

Techniques: Clinical Proteomics, Isolation, Saline, RNA Sequencing, Flow Cytometry