Figure 1
Voluntary running behavior and impact on body weight. (a) Daily running distance, (b) daily running patterns and (c) total running distance of male runners during 29 days of running. (d) Daily running distance, (e) daily running patterns and (f) total running distance of female runners during 29 days of running. (g) Body weight (BW) evolution (time × housing interaction, F(4, 104) = 20.89, P < 0.0001; Tukey post hoc: *P < 0.05, **P < 0.01, SED versus RUN) and (h) total BW gain (t = 5.158, df = 26, P < 0.0001 versus SED) of male runners (RUN) and sedentary (SED) controls. (i) BW evolution (time × housing interaction, F(4, 88) = 5.14, P = 0.0009; Sidak post hoc: *P < 0.05, SED versus RUN) and (j) total body weight gain (t = 2.646, df = 22, P = 0.0147 versus SED) of female runners (RUN) and sedentary (SED) controls. Data are presented as the mean ± S.E.M or as box plots indicating the median (line), the interquartile range, and the minimum to maximum values of the data distribution, with dots representing individual rats. a,b,c,g,h: n = 14/group; d,e,f,i,j: n = 12/group.
Figure 2
Long-term voluntary running lowers SCN ΔFOSB. (a–c) Quantification of ΔFOSB-positive cell numbers in the SCN (between bregma –0.6 mm and –0.84 mm) of (a) male runners (RUN) and sedentary (SED) controls (running for 29 days; bregma × housing interaction, F(2, 38) = 0.5658, P = 0.5726; main effect of bregma, F(1.65, 31.34) = 6.093, P = 0.0086; main effect of housing, F(1, 19) = 3.715, P = 0.069), (b) female runners (RUN) and sedentary (SED) controls (bregma × housing interaction, F(2, 40) = 0.7783, P = 0.4660; main effect of bregma, F(1.256, 25.11) = 17.55, P < 0.0001; main effect of housing, F(1, 20) = 6.586, P = 0.0184), and in the SCN of (c) an independent replication cohort of female runners (RUN) and sedentary (SED) controls (running for 28 days; bregma × housing interaction, F(2, 40) = 10.83, P = 0.0002, Sidak’s post hoc: **P = 0.0035, SED–0.72 versus RUN–0.72; main effect of bregma, F(1.912, 38.24) = 21.57, P < 0.0001; main effect of housing, F(1, 20) = 4.214, P = 0.0534). (d–e) Total average number of ΔFOSB-positive cells in the SCN (between bregma –0.6 mm and –0.84 mm) of (d) male runners (RUN) and sedentary (SED) controls (t = 1.927, df = 19, P = 0.069 versus SED), (e) female runners (RUN) and sedentary (SED) controls (t = 2.566, df = 20, P = 0.0184 versus SED), and in the SCN of (f) an independent replication cohort of female runners (RUN) and sedentary (SED) controls (t = 2.053, df = 20, P = 0.0534 versus SED). Data are presented as the mean ± S.E.M (a–c) or as box plots indicating the median (line), the interquartile range, and the minimum to maximum values of the data, with dots representing individual rats (d–f). a,d: n = 10–11/group; b,e: n = 10–12/group, c,f: n = 11/group.
Figure 3
Estrous stages modulate SCN ΔFOSB. (a) Quantification of ΔFOSB-positive cell numbers in the SCN (between bregma –0.6 mm and –0.84 mm) of female runners (RUN) and sedentary (SED) controls divided by estrous stage during collection of brain tissue (main effect of housing, F(1, 17) = 9.213, P = 0.0075; Tukey post hoc: P = 0.002, SEDDiestrus versus RUNDiestrus; P = 0.013, SEDdiestrus versus SEDproestrus). (b) Quantification of ΔFOSB-positive cell numbers in the SCN (between bregma –0.6 mm and –0.84 mm) of an independent replication cohort of female sedentary controls killed at proestrus or diestrus after completing at least two estrous cycles (t = 3.001, df = 8, P = 0.0170 versus diestrus). Data are presented as the mean ± S.E.M (a) or as box plots indicating the median (line), the interquartile range, and the minimum to maximum values of the data, with dots representing individual rats (b). (a) proestrus: n = 9–10/group; estrus: n = 4/group; metestrus: n = 3–5/group; diestrus: n = 4–5/group; (b) proestrus: n = 5; diestrus: n = 5.
Figure 4
Estradiol replacement following ovariectomy lowers SCN ΔFOSB. (a) Average fluid intake (per cage) of water containing vehicle or estradiol during the 23 days of estradiol replacement. (b) Estradiol intake (per animal), normalized for body weight, calculated based on water intake. (c) Total body weight gain after 23 days of vehicle- or estradiol treatment. (d,e) Representative uteri (d; left, vehicle; right, estradiol) and uterus weight comparison (e) of vehicle- or estradiol-treated ovariectomized rats. (f) Quantification of ΔFOSB-positive cell numbers in the SCN (between bregma –0.60 mm and –0.84 mm) of ovariectomized rats with vehicle or estradiol (E2) replacement (bregma × E2 treatment interaction, F(2, 26) = 0.9877, P = 0.9877; main effect of bregma, F(1.533,19.93) = 39.71, P < 0.0001; main effect of E2 treatment, F(1,13) = 5.629, P = 0.0338). (g) Total average number of ΔFOSB-positive cells in the SCN (between bregma –0.60 mm and –0.84 mm) following vehicle- or estradiol treatment (t = 2.373, df = 13, P = 0.0338). Data are presented as the mean ± S.E.M (d,f) or as box plots indicating the median (line), the interquartile range, and the minimum to maximum values of the data, with dots representing individual rats (b,e,g). (e) vehicle: n = 4; estradiol: n = 4, (a–c,f–g) vehicle: n = 7; estradiol: n = 8.
Figure 5
Co-expression of ΔFOSB with VIP, but not AVP, in the SCN. (ai–ii) Examples of (ai) VIP expression and (aii) AVP expression in the SCN (bregma –0.72 mm) of a female runner. (bi–ii) Insets, shown as blue squares in ai–ii, at 10x magnification for (bi) VIP and (bii) AVP. (ci–ii) ΔFOSB expression in insets, shown as blue squares in ai–ii. (di–ii) Co-expression of (di) ΔFOSB and VIP and (dii) ΔFOSB and AVP in insets, shown as blue squares in ai–ii. White arrowheads indicate co-expression of ΔFOSB and VIP. ai–ii: scale bar = 100 μm; bi–ii, ci–ii, di–ii: scale bar = 50 μm.