Environmental enrichment requires adult neurogenesis to facilitate the recovery from psychosocial stress methods
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Affective behaviors
During the final 7 days of the SC re-exposure period, we conducted a battery of behavioral tests to further elucidate the affective phenotype of Ctrl and NG- mice after either SC → EE or SC → IE. The same order of tests: exploration in a novel cage, preference for a sweet solution, anxiety, social preference and preference for the intruder/aggressor, was conducted for each group to assay affective behaviors ( ).
Materials and methods
We used adult (experimental treatments began at 8 weeks of age and continued to 20 weeks of age) transgenic mice and control littermates from heterozygote (female hGFAPtk) to wildtype (male C57BL/6J) crossings. hGFAPtk transgenic mice were generated as described below and transferred to a C57BL/6J background by >10 backcrosses. Before differential housing, animals were housed four per cage in a 12-h light-dark colony room with lights off from 0900 to 2100 h. The procedures described were conducted in accordance with the NIH guidelines and were approved by the NIMH Institutional Animal Care and Use Committee.
Drugs
Valganciclovir (VGCV, Roche, Indianapolis, IN, USA)-the L-valyl ester of ganciclovir-was administered for up to 12 weeks through the animals' chow at a concentration of 15 mg kg -1 body weight/day. After phosphorylation by HSV-tk, ganciclovir is toxic to proliferating cells in S-phase of mitosis. As control mice do not express HSV-tk, VGCV administration does not suppress proliferation of GFAP-positive cells in these animals. To control for any other effects of the drug, both control and hGFAPtk mice received the same VGCV-containing chow.
Social conflict (SC)
Repeated social defeat was used to induce a depressive-like phenotype in intruder mice during the SC induction phase. Aggressor CD-1 male mice were single-housed in a large polycarbonate cage (24.0 cm × 46.0 cm × 15.5 cm, Lab Products, Seaford, DE, USA) for 2-4 weeks with bedding incompletely refreshed once per week. Male Ctrl and NG- mice were subsequently placed into the resident CD-1 mouse's home cage into which a 1/8-inch thick perforated transparent polycarbonate partition had been placed down the middle to separate the pair. The partition allowed olfactory, visual and auditory communication but prevented tactile contact. Testing commenced after a two-day accommodation period. The partition was removed for 5 min d -1 for 14 consecutive days (between 1600 and 1800 h) to allow agonistic encounters between the mice. After eac...
Enriched housing (EE)
Enriched housing consisted of a (24.5 cm × 40.5 cm × 18.5 cm) polycarbonate cage (Tecniplast) with wood chip bedding, nesting material, running wheels and numerous polycarbonate and paper tubes of different shapes and sizes (Bio-Serv, Frenchtown, NJ, USA). To minimize stress from novel objects and to maintain a sanitary environment, polycarbonate tubes and wheels were washed with warm water, dried and replaced in the originating cage once every 7 days (see Supplementary Figure 3a for comparative cage sizes).
Reintroduction into social conflict (SC)
Following 21 days of differential housing, experimental mice were reintroduced into SC environment and co-housed with the same CD-1 mouse from the initial induction period for another 14-day period. Similar conditions to the first resident-intruder induction period were used.
Saccharine preference test
Mice were subjected to a 2-bottle choice saccharine (50 mg l -1 ) preference test. Testing was carried out over 3 days in the home cage. Each mouse was given simultaneous access to two dual ball sipper-top bottles (AnaCare, Potomac, MD, USA): one with purified Milli-Q water and one containing a 50 mg l -1 saccharine (Sigma Aldrich, St Louis, MO, USA) solution ad libitum. The amounts consumed over the whole 3-day period were measured and the preference for the saccharine solution was calculated according to the formula: Percentage of preference=((saccharine solution intake/total intake) × 100%).
Social-interaction and dominant-interaction tests
Mice were placed into an open field containing 2 upside-down wire mesh pencil cups that served both as novel objects and container for another mouse. In the social interaction test, the mouse placed in the cup was an unfamiliar mouse of same genetic background (C57BL/6J) and age. In the dominant interaction test (performed 1 day after the social interaction test), the cup contained the same aggressor that the animal had lived with under the SC conditions. Mice were placed in the middle of the open field and allowed to explore for 1 h. Sessions were videotaped and later analyzed using automated behavioral tracking software (TopScan/ObjectScan, Cleversystems, Leesburg, VA, USA). The software reliably scores the frequency and duration of sniffing on either object or mouse through identification of the animals shape including nose, body and tail. Interaction ratios were calculated u...
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Evidence quotes (8)
During the final 7 days of the SC re-exposure period, we conducted a battery of behavioral tests to further elucidate the affective phenotype of Ctrl and NG- mice after either SC → EE or SC → IE. The same order of tests: exploration in a novel cage, preference for a sweet solution, anxiety, social preference and preference for the intruder/aggressor, was conducted for each group to assay affective behaviors ( ).
We used adult (experimental treatments began at 8 weeks of age and continued to 20 weeks of age) transgenic mice and control littermates from heterozygote (female hGFAPtk) to wildtype (male C57BL/6J) crossings. hGFAPtk transgenic mice were generated as described below and transferred to a C57BL/6J background by >10 backcrosses. Before differential housing, animals were housed four per cage in a 12-h light-dark colony room with lights off from 0900 to 2100 h. The procedures described were conducted in accordance with the NIH guidelines and were approved by the NIMH Institutional Animal Care and Use Committee.
Valganciclovir (VGCV, Roche, Indianapolis, IN, USA)-the L-valyl ester of ganciclovir-was administered for up to 12 weeks through the animals' chow at a concentration of 15 mg kg -1 body weight/day. After phosphorylation by HSV-tk, ganciclovir is toxic to proliferating cells in S-phase of mitosis. As control mice do not express HSV-tk, VGCV administration does not suppress proliferation of GFAP-positive cells in these animals. To control for any other effects of the drug, both control and hGFAPtk mice received the same VGCV-containing chow.
Repeated social defeat was used to induce a depressive-like phenotype in intruder mice during the SC induction phase. Aggressor CD-1 male mice were single-housed in a large polycarbonate cage (24.0 cm × 46.0 cm × 15.5 cm, Lab Products, Seaford, DE, USA) for 2-4 weeks with bedding incompletely refreshed once per week. Male Ctrl and NG- mice were subsequently placed into the resident CD-1 mouse's home cage into which a 1/8-inch thick perforated transparent polycarbonate partition had been placed down the middle to separate the pair. The partition allowed olfactory, visual and auditory communication but prevented tactile contact. Testing commenced after a two-day accommodation period. The partition was removed for 5 min d -1 for 14 consecutive days (between 1600 and 1800 h) to allow agonistic encounters between the mice. After each interaction period, the partition was replaced with the aggressor male returned to his initial compartment; therefore experimental mice were consistently housed with the same CD-1 mouse. Interaction periods were videotaped under red light and later analyzed by an unbiased observer for aggressive (...
Enriched housing consisted of a (24.5 cm × 40.5 cm × 18.5 cm) polycarbonate cage (Tecniplast) with wood chip bedding, nesting material, running wheels and numerous polycarbonate and paper tubes of different shapes and sizes (Bio-Serv, Frenchtown, NJ, USA). To minimize stress from novel objects and to maintain a sanitary environment, polycarbonate tubes and wheels were washed with warm water, dried and replaced in the originating cage once every 7 days (see Supplementary Figure 3a for comparative cage sizes).
Following 21 days of differential housing, experimental mice were reintroduced into SC environment and co-housed with the same CD-1 mouse from the initial induction period for another 14-day period. Similar conditions to the first resident-intruder induction period were used.
Mice were subjected to a 2-bottle choice saccharine (50 mg l -1 ) preference test. Testing was carried out over 3 days in the home cage. Each mouse was given simultaneous access to two dual ball sipper-top bottles (AnaCare, Potomac, MD, USA): one with purified Milli-Q water and one containing a 50 mg l -1 saccharine (Sigma Aldrich, St Louis, MO, USA) solution ad libitum. The amounts consumed over the whole 3-day period were measured and the preference for the saccharine solution was calculated according to the formula: Percentage of preference=((saccharine solution intake/total intake) × 100%).
Mice were placed into an open field containing 2 upside-down wire mesh pencil cups that served both as novel objects and container for another mouse. In the social interaction test, the mouse placed in the cup was an unfamiliar mouse of same genetic background (C57BL/6J) and age. In the dominant interaction test (performed 1 day after the social interaction test), the cup contained the same aggressor that the animal had lived with under the SC conditions. Mice were placed in the middle of the open field and allowed to explore for 1 h. Sessions were videotaped and later analyzed using automated behavioral tracking software (TopScan/ObjectScan, Cleversystems, Leesburg, VA, USA). The software reliably scores the frequency and duration of sniffing on either object or mouse through identification of the animals shape including nose, body and tail. Interaction ratios were calculated using the interaction quotient (interaction duration on mouse/interaction duration on empty cup).
Machine-readable layer
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