Running Wheel Exercise
Objective: To assess priming effects of running wheel exercise on subsequent enriched environment exposure by comparing hippocampal effects across different temporal sequences of physical activity and environmental enrichment in female mice
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Protocol Steps
Animal acquisition and housing setup
Obtain 40 female C57BL/6 mice, 8 weeks old. Randomly distribute to four experimental groups (N = 10 per group). House all animals in same room with constant 12-hour light/dark cycle.
Note: Female mice selected to avoid confounding effects of territorial behavior and social hierarchy in males
View evidence from paper
“Forty female C57BL/6 mice, 8-weeks old at the beginning of the experiment, were obtained from Charles River (Sulzfeld, Germany). The mice were randomly distributed to four experimental groups, N = 10 per group. All animals were kept in the same room with a constant 12-h-light/dark-cycle”
Establish experimental groups
Create four experimental conditions: (1) RUNENR - running wheel in phase 1, enriched environment in phase 2; (2) RUNSTD - running wheel in phase 1, control conditions in phase 2; (3) STDENR - control conditions in phase 1, enriched environment in phase 2; (4) STDSTD - control conditions in both phases
Note: RUNSTD and RUNENR differ only in phase 2; STDENR and RUNENR differ only in phase 1
View evidence from paper
“The main experimental group consisted of mice that lived in a cage equipped with a running wheel during the first phase, followed by the exposure to an enriched environment in the second (RUNENR). The first control condition consisted of mice that were running in the first phase but lived under control conditions in the second (RUNSTD). STDENR thus differed only in the first phase, not the second. All three groups were compared to mice living under control conditions (STDSTD).”
Phase 1: First 10-day period
Implement first experimental phase lasting 10 days. RUNENR and RUNSTD groups have access to running wheel. STDENR and STDSTD groups remain under control conditions.
Note: Running wheel access is the primary manipulation in phase 1
View evidence from paper
“The experimental period lasted 45 days, divided into a first phase of 10 days”
BrdU administration at end of phase 1
At end of first 10-day period, administer single daily injections of BrdU (50 μg/g body weight in 0.9% saline) to five mice from each group for 3 consecutive days
Note: BrdU labels progenitor cells generated in last 3 days of phase 1 to track survival-promoting effects of enrichment in phase 2
View evidence from paper
“At the end of the first period, five mice of each group received single daily injections of persistent S-phase label bromodeoxyuridine (BrdU; 50 μg/g body weight in 0.9% saline; Sigma) for 3 days”
Phase 2: Second 35-day period
Implement second experimental phase lasting 35 days. RUNENR group exposed to enriched environment. RUNSTD group remains under control conditions (discontinuation of running wheel). STDENR group exposed to enriched environment. STDSTD group remains under control conditions.
Note: Running wheel access is discontinued for RUNSTD group; measured effects represent sustained effects after 35 days of discontinuation
View evidence from paper
“a second phase of 35 days. the measured effects of running in this condition represented sustained effects after 35 days of discontinuation of physical exercise”
End-point analysis
Analyze all animals at end of 45-day experimental period (10 days phase 1 + 35 days phase 2)
Note: Remaining five mice per group were intended for gene expression study which failed technically
View evidence from paper
“Because all animals were analyzed at the end of the 45 days. The remaining animals were intended for a gene expression study, which failed technically.”