Journal List > Korean J Physiol Pharmacol > v.13(5) > 1025623

Hong, Song, Yun, Cho, and Chung: Increase of NADPH-diaphorase Expression in Hypothalamus of Stat4 Knockout Mice

Abstract

Signal transducer and activator of transcription 4 (STAT4), a STAT family member, mediates interleukin 12 (IL12) signal transduction. IL12 is known to be related to calorie-restricted status. In the central nervous system, IL12 also enhances the production of nitric oxide (NO), which regulates food intake. In this study, the expression of neuronal NO synthase (Nos1), which is also related to food intake, was investigated in the hypothalamic areas of Stat4 knockout (KO) mice using nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry, a marker for neurons expressing Nos1 enzyme. Western blots were also performed to evaluate Nos1 and Fos expression. Wild-type Balb/c (WT group, n=10 male) and Stat4 KO mice (Stat4 KO group, n=8 male) were used. The body weight and daily food intake in the WT group were 22.4±0.3 and 4.4 g per day, while those in the Stat4 KO group were 18.7±0.4 and 1.8 g per day, respectively. Stat4 mice had lower body weight and food intake than Balb/c mice. Optical intensities of NADPH-d-positive neurons in the paraventricular nucleus (PVN) and lateral hypothalamic area (LHA) of the Stat4 KO group were significantly higher than those of the WT group. Western blotting analysis revealed that the hypothalamic Nos1 and Fos expression of the Stat4 KO group was up-regulated, compared to that in the WT group. These results suggest that Stat4 may be related to the regulation of food intake and expression of Nos1 in the hypothalamus.

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Fig. 1.
Body weight in wild-type Balb/c and Stat4 knockout (KO) mice. Values are mean±SEM. p<0.05 vs. Balb/c mice.
kjpp-13-337f1.tif
Fig. 2.
Daily food intake in wild-type Balb/c and Stat4 knockout (KO) mice. Values are mean±SEM. p<0.05 vs. Balb/c mice.
kjpp-13-337f2.tif
Fig. 3.
Distribution of NADPH-diaphorase-positive neurons in the hypothalamic regions. (A) The paraventricular nucleus of Balb/c mice (WT group). (B) The paraventricular nucleus of Stat4 knockout mice (Stat4 KO group). (C) The lateral hypothalamic area of the WT group. (D) The lateral hypothalamic area of the Stat4 KO group. Scale bar represents 80 μm.
kjpp-13-337f3.tif
Fig. 4.
Optical density of NADPH-diaphorase-positive neurons in the hypothalamic regions. Values are mean±SEM. NADPH-d, nicotinamide adenine dinucleotide phosphate-diaphorase; PVN, paraventricular nucleus; LHA, lateral hypothalamic area; Stat4 KO, Stat4 knockout mice. p<0.05 vs. Balb/c mice.
kjpp-13-337f4.tif
Fig. 5.
Western blotting analysis of Nos1 and Fos immunoreactivity. (A) Immunoreactive bands on western blots. (B) Expression of Nos1 and Fos proteins was calculated by densitometer and normalized to that of Actb (internal control). Values are mean±SEM. Nos1, nitric oxide synthase 1, neuronal; Fos, FBJ osteosarcoma oncogene; Actb, actin, beta; Stat4 KO, Stat4 knockout mice. p<0.05 vs. Balb/c mice.
kjpp-13-337f5.tif
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