The effects of repeated restraint stress on energy balance and behavior of mice with selective deletion of CRF receptors

Figures & data

Table I.  PCR Primers for genotyping.

CRFR1 Mice: Wild type amplification
Wild type primer	5′- TCT CAG GAT TGC TAA GTT CAG -3′
Common primer	5′- TCT CAG GAT TGC TAA GTT CAG -3′
CRFR1 Mice: Knockout amplification
Knockout primer	5′- AAC TTC CTG ACT AGG GG -3′
Anti-sense primer	5′- TCT CAG GAT TGC TAA GTT CAG -3′
CRFR2 Mice
Knockout primer	5′- GGG AAC TTC CTG ACT AGG GG -3′
Wild type primer	5′- TGT GGA AGC TGA TCC TGG TG -3′
Anti-sense primer	5′ CCT GAT GCC TGC TGA GTT GA -3′

Table II.  Body weight and food intakes.

	Start body weight (g/mouse)	End body weight (g/mouse)	Total food intake (g/10 days)
WT Control	24.8 ± 0.4^B	25.7 ± 0.4^B	40.4 ± 1.1
WT RRS	24.8 ± 0.4^B	25.3 ± 4^B	42.3 ± 1.6
ADX Control	22.2 ± 0.4^A	22.7 ± 0.6^A	38.2 ± 1.2
ADX RRS	22.0 ± 0.8^A	22.1 ± 0.8^A	40.3 ± 0.8
CRFR1-KO Control	25.8 ± 0.6^B	26.1 ± 0.5^B	41.3 ± 0.8
CRFR1-KO RRS	25.8 ± 0.4^B	26.6 ± 0.4^B	43.1 ± 1.2
CRFR2-KO Control	24.7 ± 0.5^B	26.7 ± 0.5^B	40.4 ± 1.2
CRFR2-KO RRS	24.7 ± 0.7^B	24.8 ± 0.8^B	41.1 ± 1.5

Data are mean ± SEM for groups of 9–22 mice. Values for start or end weight that do not share a common superscript are significantly different at P < 0.05 and indicate differences between treatment groups. Start weight is the weight of the mice immediately before the first RRS. End weight is weight of the mice immediately before acclimation to the EPM, 8 days after the end of RRS. ADX, adrenalectomised, and corticosterone replacement; CRFR, corticotropin releasing factor receptor; KO, gene deletion; RRS, daily restraint stress for 3 days; WT, wild type.

Figure 1.  Change in body weight from the morning before the first restraint (day 0). Data are mean ± SEM for groups of 9 to 22 mice. The WT mice represent combined groups of WT from CRFR1-KO and CRFR2-KO colonies. Asterisks indicate days on which there was a significant difference in weight change between Control and RRS mice. ADX, adrenalectomised and corticosterone replacement; CRFR, corticotropin releasing factor receptor; EPM, elevated plus maze exposure; KO, gene deletion; L/D, light/dark choice test; RRS, daily restraint stress for 3 days; WT, wild type.

Figure 2.  Food intake of the mice represented as total intake for 3 days before, during, and after RRS. The asterisk indicates a significant difference (P < 0.05) in intake (g/mse[mouse]) of Control versus RRS mice. Data are mean ± SEM for groups of 9–22 mice. ADX, adrenalectomised and corticosterone replacement; CRFR, corticotropin releasing factor receptor; KO, gene deletion; RRS, daily restraint stress for 3 days; WT, wild type.

Figure 3.  Behavior measured during 5 min in the elevated plus maze (EPM) on day 11 of the experimental period, 9 days after the end of daily RRS for 3 days (RRS). Values on any specific axis that do not share a common superscript are significantly different at P < 0.05. Data are mean ± SEM for groups of 9–22 mice. ADX, adrenalectomised and corticosterone replacement; CRFR, corticotropin releasing factor receptor; KO, gene deletion; WT, wild type.

Figure 4.  Behavior measured in the light/ dark (L/D) choice test on day 14 of the experimental period, 12 days after the end of daily restraint for 3 days (RRS). Values on an axis that do not share a common superscript are significantly different at P < 0.05. Data are mean ± SEM for groups of 9–22 mice. ADX, adrenalectomised and corticosterone replacement; CRFR, corticotropin releasing factor receptor; KO, gene deletion; WT, wild type.

Figure 5.  Serum corticosterone measured before (time 0) and 25 min (time 25) after the start of testing in the EPM on day 11 of the experimental period. Values that do not share a common superscript are significantly different at P < 0.05. ADX, adrenalectomised and corticosterone replacement; CRFR, corticotropin releasing factor receptor; CTRL, control; KO, gene deletion; RRS, daily restraint stress for 3 days; WT, wild type.