Journal: Neuropsychopharmacology

Article Title: Hypocretin in the nucleus accumbens shell modulates social approach in female but not male California mice

doi: 10.1038/s41386-024-01937-9

Figure Lengend Snippet: In male Mus , NAc cells were organized into 8 cell types: astrocytes (Astro), oligodendrocytes (Oligo), endothelial cells (Endo), interneurons (IN), dopamine receptor 1-expressing cells (D1), and dopamine receptor 2-expressing cells (D2). Hcrtr2 is primarily expressed in the dopamine D1 receptor medium spiny neurons and interneurons, whereas Hcrtr1 is primarily expressed in the D1 and D2 medium spiny neurons ( A ). A Dotplot shows that Hcrtr1 and Hcrtr2 have distinct expression patterns across all cell types in male Mus . Hcrtr1 has the highest average and percent expression in the interneurons, whereas Hcrtr2 has the highest average and percent expression in D1 neurons ( B ). In the nucleus accumbens but not medial prefrontal cortex or ventral tegmental area Hcrtr2 was more abundant than Hcrtr1 ( C ). Timeline of experiment in stressed female California mice and schematic of the mechanism of action for selective HcrtR2 antagonist TSC OX2 29 ( D ). Infusion of TSC OX2 29 in the NAcSh of female California mice previously exposed to social defeat stress increased social approach ( E ) but had no effects on social vigilance behavior ( F ), behaviors during the acclimation phase ( G ). * p < 0.05, + p = 0.06 vs. saline. *** p < 0.001 vs. Hcrtr1 . Group n’s: gene expression; male/control n = 7, male/stress n = 6, female/control n = 8, female/stress n = 7, pharmacology; female/saline n = 9, female/OX2 29 n = 9.

Article Snippet: Mice were implanted with guides aimed at the NAcSh and one week later received either saline (vehicle) or 13 μg HcrtR2 antagonist (TCS OX2 29, Tocris #3371) before behavior testing.

Techniques: Expressing, Saline, Gene Expression, Control

Journal: Biological psychiatry

Article Title: Hypocretin/orexin signaling in the hypothalamic paraventricular nucleus is essential for the expression of nicotine withdrawal.

doi: 10.1016/j.biopsych.2011.06.025

Figure Lengend Snippet: Figure 1. Hypocretin transmission through the specific activation of hypocretin receptor-1 modulates the expression of the nicotine withdrawal syndrome. Acute mecamylamine (2 mg/kg, subcutaneous) was administered to precipitate nicotine withdrawal in C57BL/6J mice receiving a continuous perfusion of nicotine (25 mg/kg/day) from subcutaneously implanted osmotic minipumps. SB334867 and TCSOX229 were administered 30 minutes before mecamyl- amine. (A,B,C) A global withdrawal score was calculated for each animal by giving each individual sign a relative weight in (A) wild-type or preprohypocretin knockout (n 9–14 mice per group) mice, in (B) vehicle or SB334867 (5 and 10 mg/kg, intraperitoneal [IP]) pretreated C57BL/6J mice (n 9–24 mice per group), and in (C) vehicle or TCSOX229 (5 and 10 mg/kg, IP) pretreated C57BL/6J mice (n 8–12 mice per group). (D) Schematic anatomic representation of lateral hypothalamic area subdivisions adapted from Paxinos and Franklin (60) stereotaxic atlas. Colored areas delineate regions where c-Fos expression was examined. (E,F) Percentage of c-Fos-positive hypocretin-1-expressing neurons in vehicle, SB334867 (5 mg/kg, IP), or TCSOX229 (10 mg/kg, IP) pretreated C57BL/6J mice (n 6–13 mice per group) in the perifornical and dorsomedial hypothalamus (PFA/DMH) and in the lateral hypothalamus. (G) Representative images of sections of the PFA/DMH obtained by confocal microscopy after direct double labeling combining rabbit polyclonal antiserum to c-Fos (red) with mouse monoclonal antibody to hypocretin-1 (green). Arrowheads indicate c-Fos-positive hypocretin-1-expressing neurons. Scale bar, 100 m. Data are expressed as mean SEM. p .05, p .01 compared with the control group; #p .05, ##p .01 comparison between pretreatments or genotypes. KO, knockout; LH, lateral hypothalamus; NIC, nicotine; SAL, saline; SB, SB334867; TCS, TCSOX229; VEH, vehicle; WT, wild-type; PFA/DMH, perifornical area and dorsomedial hypothalamus. (Image in (D) reproduced with permission from Paxinos and Franklin (60), copyright Elsevier, 1997).

Article Snippet: The Hcrtr-2 antagonist TCSOX229 (5 and 10 mg/kg) (Tocris) was dissolved in physiological saline (.9% NaCl) and administered by IP route in a volume of 5 mL/kg body weight.

Techniques: Transmission Assay, Activation Assay, Expressing, Knock-Out, Confocal Microscopy, Labeling, Control, Comparison, Saline

Journal: Biological psychiatry

Article Title: Hypocretin/orexin signaling in the hypothalamic paraventricular nucleus is essential for the expression of nicotine withdrawal.

doi: 10.1016/j.biopsych.2011.06.025

Figure Lengend Snippet: Figure 2. Hypocretins acting on hypocretin receptor-1 regulate the activation of the hypothalamic paraventricular nucleus associated with nicotine withdrawal. (A,B,C) c-Fos expression in the PVN 2 hours after precipitation of nicotine withdrawal by acute mecamylamine (2 mg/kg, subcutaneous) injection in(A)wild-typeorpreprohypocretinknockoutmice(n5–6micepergroup),in(B)vehicleorSB334867(5mg/kg,IP)pretreatedC57BL/6Jmice(n5–8mice per group), and in (C) vehicle or TCSOX229 (10 mg/kg, IP) pretreated C57BL/6J mice (n 7–9 mice per group). SB334867 and TCSOX229 were administered 30 minutes before mecamylamine. (D,E) Representative images of the PVN obtained by confocal microscopy after direct labeling with rabbit polyclonal antiserum to c-Fos in SB334867 pretreated and knockout mice. Scale bar, 100 m. (F) Schematic anatomic representation of the PVN adapted from Paxinos and Franklin (60) stereotaxic atlas. The colored area delineates the region where c-Fos expression was examined. (G,H) Percentage of CRF (G) and arginine-vasopressin (H) cells expressing c-Fos in the PVN 2 hours after precipitation of nicotine withdrawal by mecamylamine (2 mg/kg, subcutaneous) in vehicle or SB334867 (5 mg/kg, IP) pretreated C57BL/6J mice (n 3–6 mice per group). Representative images of c-Fos expressing CRF neurons in the PVN are showninFigureS4inSupplement1.Dataareexpressedasmean SEM. p.05, p.01comparedwiththecontrolgroup; #p.05, ##p.01comparison between pretreatments or genotypes. AVP, arginine-vasopressin; CRF, corticotrophin-releasing factor; IP, intraperitoneal; KO, knockout; NIC, nicotine; PVN, paraventricular nucleus; SAL, saline; SB, SB334867; TCS, TCSOX229; VEH, vehicle; WT, wild-type. (Image in (F) reproduced with permission from Paxinos and Franklin (60), copyright Elsevier, 1997).

Article Snippet: The Hcrtr-2 antagonist TCSOX229 (5 and 10 mg/kg) (Tocris) was dissolved in physiological saline (.9% NaCl) and administered by IP route in a volume of 5 mL/kg body weight.

Techniques: Activation Assay, Expressing, Injection, Confocal Microscopy, Labeling, Knock-Out, Saline