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1.
Fig. 6

Fig. 6. From: RGS2 is a feedback inhibitor of melatonin production in the pineal gland.

Schematic presentation of negative regulation of melatonin synthesis by RGS2. RGS2 is induced by the daily release of norepinephrine which binds to α1b- and β1-adrenergic receptors and activates adenylyl cyclase. The resulting increase in cAMP leads to the transcription of several genes including Rgs2. Translated RGS2 protein inhibits G-protein signaling thereby attenuating cAMP-mediated transcription.

Masahiro Matsuo, et al. FEBS Lett. ;587(9):1392-1398.
2.
Fig. 3

Fig. 3. From: RGS2 is a feedback inhibitor of melatonin production in the pineal gland.

RGS2 inhibits melatonin production in a dose-dependent manner. (A) Western blot of cell lysates infected with serial titers of RGS2-coding adenovirus; 1.1 to 90 virus particles per cell (MOI). The increase in pineal RGS2 protein was titer-dependent. (B) Virus titer-dependent decrease of melatonin production in pineal cells. Although no significant decrease of melatonin was found up to 10 virus particles per cell, a significant and titer-dependent decrease by RGS2 was found at 30 and 90 virus particles per cell (P < 0.05, one-way ANOVA, Control vs RGS2 infected cells at same virus concentration, n = 3 each). For further details see Section 2.

Masahiro Matsuo, et al. FEBS Lett. ;587(9):1392-1398.
3.
Fig. 4

Fig. 4. From: RGS2 is a feedback inhibitor of melatonin production in the pineal gland.

RGS2 inhibits norepinephrine- and isoproterenol-dependent elevation of N-acetylserotonin, melatonin and cAMP in pinealocyte culture media. Pineal cells were stimulated by a range of concentrations of norepinephrine (A and C) or isoproterenol (B and D). Significant inhibition of melatonin and N-acetylserotonin production by RGS2 occurred at concentrations of 10−9–10−6 and 10−8–10−6 M norepinephrine, respectively (A and C, P < 0.05, one-way ANOVA, Control vs RGS2-infected cells at the same concentration of norepinephrine, n = 3 each). Significant inhibition of both melatonin and N-acetylserotonin production by RGS2 was found at the concentrations of 10−9–10−8 M isoproterenol (B and D, P < 0.05, one-way ANOVA, Control vs RGS2-infected cells at the same concentration of isoproterenol, n = 3 each). (E) Norepinephrine stimulation of cAMP content of media was decreased in cultures of cells in which RGS2 was overexpressed (P < 0.05, one-way AVNOVA, Control vs RGS2-infected cells at 10−5 µM norepinephrine, n = 3 each). For further details see Section 2.

Masahiro Matsuo, et al. FEBS Lett. ;587(9):1392-1398.
4.
Fig. 2

Fig. 2. From: RGS2 is a feedback inhibitor of melatonin production in the pineal gland.

Increase of Rgs2 transcripts and RGS2 protein following treatment with norepinephrine or a cAMP analog. (A) Rgs2 transcription was induced by 1 µM norepinephrine application, and reached its peak 2 h after stimulation. Significant induction of Rgs2 was found 1 to 4 h after stimulation (P < 0.05, one-way ANOVA vs expression level at 0 h, n = 3 each). (B) RGS2 protein increased within 2 h by treatment with 1 µM norepinephrine (P < 0.05, one-way ANOVA vs expression level at 0 h, n = 3 each). Inset shows a representative Western blot result of RGS2 protein expression following norepinephrine stimulation. (C) Application of the cAMP analog, dibutyryl–cAMP, induced transcription of Rgs2 (Control vs DBcAMP, 4.3 ± 0.55% vs 100.0 ± 9.41%; P < 0.05, two-tailed t-test, n = 3 each). For further details see Section 2.

Masahiro Matsuo, et al. FEBS Lett. ;587(9):1392-1398.
5.
Fig. 1

Fig. 1. From: RGS2 is a feedback inhibitor of melatonin production in the pineal gland.

Transcript levels of RGS family members in pineal glands obtained during the day or night. (A) Transcript levels of 17 RGS members in pineal gland either at day (Zeitgeber time; ZT7, white box) or night (ZT19, black box) are shown. Rgs2, Rgs4 and Rgs20 exhibited significantly different expression levels between ZT7 and ZT19 (ZT7 vs ZT19 of Rgs2: 1.3 ± 0.12 vs 28.9 ± 6.28; Rgs4: 11.4 ± 1.32 vs 4.7 ± 0.23; Rgs20: 26.9 ± 3.27 vs 13.4 ± 1.28, P < 0.05, two-tailed t-test, n = 3 each). Note Rgs2 was the only molecule that had higher expression at ZT19. (B and C) Levels of Rgs2 mRNA or RGS2 protein expression are shown relative to their peak at ZT19. Gray shading indicates time when lights were off (night time), and the expression value at ZT7 is plotted twice for presentation. For further details see Section 2.

Masahiro Matsuo, et al. FEBS Lett. ;587(9):1392-1398.
6.
Fig. 5

Fig. 5. From: RGS2 is a feedback inhibitor of melatonin production in the pineal gland.

RGS2 inhibits the norepinephrine-dependent increase of the abundance of Aanat, Dio2 and Fosl2 transcripts. The effect of RGS2 overexpression on the abundance of several transcripts was determined following by a 2-h application of 1 µM norepinephrine. Norepinephrine did not increase the abundance of Tph1 mRNA (A). However, a greater than 5-fold increase was observed for Aanat (B), Dio2 (C) and Fosl2 (D). Increased transcript levels in RGS2-infected cells was decreased to 47.7% for Aanat, to 47.1% for Dio2, and to 66.0% for Fosl2 (P < 0.05, one-way ANOVA, n = 3 each) compared to norepinephrine-stimulated control cells. For further details see Section 2. Fig. 6. Schematic presentation of negative regulation of melatonin synthesis by RGS2. RGS2 is induced by the daily release of norepinephrine which binds to α1band β1-adrenergic receptors and activates adenylyl cyclase. The resulting increase in cAMP leads to the transcription of several genes including Rgs2. Translated RGS2 protein inhibits G-protein signaling thereby attenuating cAMP-mediated transcription.

Masahiro Matsuo, et al. FEBS Lett. ;587(9):1392-1398.

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