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Research ArticleNegative Results, Disorders of the Nervous System

Validity Assessment of 5 Day Repeated Forced-Swim Stress to Model Human Depression in Young-Adult C57BL/6J and BALB/cJ Mice

Joram D. Mul, Jia Zheng and Laurie J. Goodyear
eNeuro 12 December 2016, 3 (6) ENEURO.0201-16.2016; https://doi.org/10.1523/ENEURO.0201-16.2016
Joram D. Mul
1Section on Integrative Physiology and Metabolism, Joslin Diabetes Center, Harvard Medical School, Boston, Massachusetts 02215
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Jia Zheng
1Section on Integrative Physiology and Metabolism, Joslin Diabetes Center, Harvard Medical School, Boston, Massachusetts 02215
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Laurie J. Goodyear
1Section on Integrative Physiology and Metabolism, Joslin Diabetes Center, Harvard Medical School, Boston, Massachusetts 02215
2Department of Medicine, Brigham, and Women’s Hospital, Harvard Medical School, Boston, Massachusetts 02215
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    Figure 1.

    5d-RFSS paradigm. Experimental timeline of 5d-RFSS in C57BL/6J or BALB/cJ mice. SPT1 and TST1 were performed on days −3 to 0, before stressed mice underwent 10 min of forced-swim stress during 5 consecutive days (days 1–5; induction phase), followed by SPT2 (days 5–8), SPT3 (days 22–25), an OFT (day 36), the last forced swim session (test phase; day 37), TST2 (day 38), and SPT4 (days 38–41). Nonstressed controls were not forced to swim during the induction phase but otherwise underwent the same protocol.

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    Figure 2.

    Effects of 5d-RFSS on depressive-like behavior in C57BL/6J mice. A, Immobility time of 5d-RFSS C57BL/6J mice during the induction phase (days 1–5) forced-swim tests (FST; main effect of time, F(4,36) = 8.68, p = 0.00005; post hoc test, §p = 0.006, §§p = 0.001, §§§p = 0.0002 vs day 1), and of 5d-RFSS and nonstressed control mice during the test phase (day 37; t(1,18) = 3.44, *p = 0.003 vs nonstressed mice). B, Two-bottle SPTs before 5d-RFSS (SPT1) and after 5d-RFSS (SPT2 to SPT4). C, TST immobility time before 5d-RFSS (TST1) and after 5d-RFSS (TST2; main effect of time, F(1,18) = 9.14, p = 0.007; post hoc test, §p = 0.007 vs TST1). D–F, Distance traveled (t(1,18) = 0.43, p = 0.67; D), number of center zone entries (t(1,18) = 1.01, p = 0.33; E), and number of feces produced during the OFT (t(1,18) = 0.63, p = 0.53; F). G, Change in body weight (%) from day −3 to 36 (main effect of time, F(6,108) = 90.48, p < 0.00001; post hoc test, §p = 0.02, §§p = 0.0001 vs day −3). H, I, Food intake during induction phase (days 1–5; t(1,17) = 0.77, p = 0.45; H) and during days 8–36 (t(1,18) = 0.71, p = 0.49; I). NS, Not significant. n = 10/group for all experiments.

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    Figure 3.

    Effects of 5d-RFSS on depressive-like behavior in BALB/cJ mice. A, Immobility time of 5d-RFSS BALB/cJ mice during the induction phase (days 1–5) of forced swim tests (FST; main effect of time, F(4,36) = 6.29, p = 0.0006; post hoc test, §p = 0.02, §§p = 0.002 vs day 1) and of 5d-RFSS and nonstressed control mice during the test phase (day 37; t(1,18) = 3.08, *p = 0.007). B, Two-bottle SPTs before 5d-RFSS (SPT1) and after 5d-RFSS (SPT2 to SPT4; main effect of time, F(3,54) = 12.67, p < 0.00001; post hoc test, §p = 0.002, §§p = 0.0002 vs SPT1). C, TST immobility time before 5d-RFSS (TST1) and after 5d-RFSS (TST2; main effect of time, F(1,18) = 5.06, p = 0.04; post hoc test, §p = 0.04 vs TST1). D–F, Distance traveled (t(1,18) = 0.74, p = 0.47; D), number of center zone entries (E), and number of feces produced during the OFT (t(1,18) = 0.79, p = 0.44; F). G, Change in body weight (%) from days −3 to 36 (main effect of time, F(6,108) = 36.96, p < 0.00001; post hoc test, §p = 0.002, §§p = 0.0001 vs day −3). H, I, Food intake during the induction phase (days 1 – 5; t(1,18) = 0.07, p = 0.94; H) and during days 8–36 (t(1,18) = 1.36, p = 0.19; I). NS, Not significant; ND, not detected. n = 10/group for all experiments.

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    Figure 4.

    CSDS induces a depressive-like state in C57BL/6J mice. A, CSDS mice had greater weight gain compared with nondefeated control (CON) mice (days 1 – 11; t(1,14) = 2.28, *p = 0.039). B, CON mice did not show a significant decrease in sucrose preference during two-bottle SPT2 compared with SPT1 (t(1,7) = 1.96, p = 0.1). CSDS mice demonstrated lower sucrose preference during SPT2 compared with SPT1 (before CSDS; t(1,7) = 3.16, *p = 0.016). C, CSDS mice spent less time in the interaction zone with target present during the social interaction test on day 11 (t(1,14) = 5.04, *p = 0.00018). D, Locomotor activity did not differ significantly between experimental groups during OFT on day 12 (t(1,14) = 1.01, p = 0.33). E, CSDS mice enter OFT center zone less than CON mice (t(1,14) = 2.99, *p = 0.01). NS, Not significant. n = 8/group for all experiments.

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    Figure 5.

    Effects of VWR on the persistence of immobility behavior following 5d-RFSS. A, Daily VWR distance (left; F(27,243) = 5.89, p < 0.00001) and cumulative VWR distance (right) of C57BL/6J mice. B, VWR (28 d) following 5d-RFSS lowered immobility scores during the test day swim session toward levels of nonstressed SED mice (F(2,27) = 18.45, p = 0.00001; post hoc test, *p = 0.00013 vs nonstressed SED mice; +p = 0.0034 vs nonstressed SED; §p = 0.059 vs 5d-RFSS SED mice). C, Daily VWR distance (left; F(27,243) = 4.37; p < 0.00001) and cumulative VWR distance (right) of BALBc/J mice. D, VWR (28 d) following 5d-RFSS normalized immobility scores during the test day swim session to levels of nonstressed SED mice (F(2,27) = 4.41, p = 0.02; post hoc test, *p = 0.026 vs nonstressed SED, §p = 0.068 vs 5d-RFSS SED). n = 10/group for all experiments.

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Validity Assessment of 5 Day Repeated Forced-Swim Stress to Model Human Depression in Young-Adult C57BL/6J and BALB/cJ Mice
Joram D. Mul, Jia Zheng, Laurie J. Goodyear
eNeuro 12 December 2016, 3 (6) ENEURO.0201-16.2016; DOI: 10.1523/ENEURO.0201-16.2016

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Validity Assessment of 5 Day Repeated Forced-Swim Stress to Model Human Depression in Young-Adult C57BL/6J and BALB/cJ Mice
Joram D. Mul, Jia Zheng, Laurie J. Goodyear
eNeuro 12 December 2016, 3 (6) ENEURO.0201-16.2016; DOI: 10.1523/ENEURO.0201-16.2016
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Keywords

  • anhedonia
  • animal model
  • depression
  • forced swimming
  • stress
  • voluntary wheel running

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