Fig 2: Cellular markers under basal thermoneutral naive and after heat stress. Immunoreactive-cell numbers and representative immunohistochemical images of HSP-70, GR, AVP, BDNF, NPY, COX-2 and Iba-1 positive cells within the CA1, CA3 and the DG of the hippocampus and the hypothalamic PVN (a–e). Results displayed as mean ± S.E.M.

Fig 3: Comparison of NPY levels in brain tissue of each group of mice.

Fig 4: Distribution of DßH-(green; labelled with FITC) and GAL- (A–C), L-ENK- (D–F), nNOS- (G–I), NPY- (J–L) or SOM- (M–O) positive (red; labelled with CY3) nerve fibers in the urinary bladder wall in the normal (A,D,G,J,M), RTX-treated (B,E,H,K,N), or TTX-treated (C,F,I,L,O) pigs. Red and green channels were digitally superimposed. Double-labelled fibers are yellow to orange and most of them are indicated with arrows; muscle layer (mL), blood vessel (bv), submucosa (s), urothelium (u); 20× (A–O); 40× (I).

Fig 5: The NPY-Y1 receptor and the ORX receptors are necessary for the anxiolytic effects of modafinil: immunoreactivity of orexin-A, orexin-B and NPY in the hypothalamus. Rats were killed and their brains were removed for immunoreactivity analysis 2 h following the modafinil or vehicle injunctions, as depicted in the figure. (1) Quantitative morphometric analysis of ORX-A (a–d), ORX-B (e–h) and NPY (i–l) immunoreactivity in fibers and cells of the PVN (a, e, i), ARC (b, f, j), PVTN (c, g, k) and PeF (d, h, l) in rats treated with vehicle (n=4), mifepristone (n=4), BIBO3304 (n=4) or almorexant (n=4), exposed to predator-scent stress (PSS), and then, 30 min later, treated with modafinil. (2) Representative images of ORX-A and NPY immunoreactivity in the PVTN of PSS-exposed rats treated with either vehicle (i), BIBO3304 (ii), mifepristone (iii) or almorexant (iv) before modafinil injection. Images were acquired at a × 10 magnification. Scale bar, 200 µm. The cells in green are NPY-positive and the cells in red are ORX-A-positive. Almorexant and BIBO3304 downregulated the immunoreactivity of ORX-A, ORX-B and NPY fibers and cells in the PVN (F(3,12)=61.1, P<0.0001; F(3,12)=25.4, P<0.0002; and F(3,12)=29.9, P<0.000, respectively), ARC (F(3,12)=8.7, P<0.0025; F(3,12)=26.3, P<0.0001; and F(3,12)=9.3, P<0.002, respectively), PVTN (F(3,12)=13.8, P<0.0004; F(3,12)=10.2, P<0.0015; and F(3,12)=9.2, P<0.002, respectively) and PeF (F(3,12)=3.5, P<0.05; F(3,12)=16.9, P<0.00015; and F(3,12)=9.5, P<0.002, respectively). BIBO, BIBO3304; Mifep, mifepristone; Almor, almorexant; PVN, paraventricular nucleus; ARC, arcuate hypothalamic nucleus; PVTN, paraventricular thalamic nucleus; PeF, perifornical reign of lateral hypothalamus; ORX, orexin; NPY, neuropeptide Y. Bars represent group means±s.e.m.