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1.
FIGURE 3.

FIGURE 3. From: Leptin Increases Adult Hippocampal Neurogenesis in Vivo and in Vitro.

Effect of leptin administration on body weight. Body weight was measured for 14 days during the treatment with leptin (1 mg/kg, intraperitoneal). Data are expressed as mean ± S.E. **, p < 0.01; n = 6 per group.

Jacob C. Garza, et al. J Biol Chem. 2008 June 27;283(26):18238-18247.
2.
FIGURE 4.

FIGURE 4. From: Leptin Increases Adult Hippocampal Neurogenesis in Vivo and in Vitro.

Expression of the long form leptin receptor (LepRb) in adult hippocampal stem/progenitor cells. A, RT-PCR using cDNA extracted from adult stem/hippocampal stem cells produced a 572-bp band corresponding to the C-terminal region of LepRb. B, confocal microscopic images showing that adult hippocampal stem/progenitor cells stained with Nestin (green) are positive for immunoreactivity of the leptin receptor (red). 4′,6-Diamidino-2-phenylindole staining (blue) revealing nuclei. Scale bar = 10 μm.

Jacob C. Garza, et al. J Biol Chem. 2008 June 27;283(26):18238-18247.
3.
FIGURE 7.

FIGURE 7. From: Leptin Increases Adult Hippocampal Neurogenesis in Vivo and in Vitro.

Effect of leptin on phosphorylation of Akt, STAT3, and ERK1/2 in cultured adult hippocampal stem/progenitor cells. Cells were stimulated with leptin (0.3 nm and 3 nm) or vehicle for 15 min. The phosphorylation of Akt, STAT3, and ERK1/2 was detected using phospho-specific antibodies. A, representative immunoblot showing phosphorylated Akt and total Akt. B, the ratio of densitometric measurements of phospho-Akt and total Akt expressed as percent of control. C, representative immunoblot showing phosphorylated STAT3 and total STAT3. D, the ratio of densitometric measurements of phospho-STAT3 and total STAT3 expressed as percent of control. E, representative immunoblot showing phosphorylated ERK1/2 and total ERK1/2. F, the ratio of densitometric measurements of phospho-ERK1/2 and total ERK1/2 expressed as percent control. **, p < 0.01 as compared with control. The data are representative of two individual experiments.

Jacob C. Garza, et al. J Biol Chem. 2008 June 27;283(26):18238-18247.
4.
FIGURE 6.

FIGURE 6. From: Leptin Increases Adult Hippocampal Neurogenesis in Vivo and in Vitro.

Effect of leptin treatment on differentiation of cultured adult hippocampal stem/progenitor cells. Cells that were treated with leptin (1 nm) for 48 h and labeled with BrdUrd (10 μm) in the last 4 h of incubation were allowed to differentiate for 8 days before fixation for immunohistochemical processing. A, representative microscopic images showing that BrdUrd-labeled cells differentiated into neuronal (TuJ1-positive in red) or glial (GFAP-positive in green) cells. B, quantitative analysis indicating that the percentage of BrdUrd-labeled cells that were positive for TuJ1 or GFAP was not altered by leptin treatment. Data are expressed as mean ± S.E. Scale bar = 10 μm.

Jacob C. Garza, et al. J Biol Chem. 2008 June 27;283(26):18238-18247.
5.
FIGURE 9.

FIGURE 9. From: Leptin Increases Adult Hippocampal Neurogenesis in Vivo and in Vitro.

Effect of inhibition of STAT3 on leptin-induced cell proliferation. Cultured adult hippocampal stem/progenitor cells were incubated with various concentrations of cucurbitacin I (0.1-10 nm) for 4 h followed by treatment with leptin (1 nm) for 48 h and labeling with BrdUrd (10 μm) during the last 4 h of incubation. A, quantitative analysis revealed that pretreatment with cucurbitacin I attenuated the leptin-induced increase in the number of BrdUrd-labeled cells. B, representative microscopic images of BrdUrd-labeled cells from three treatment conditions: control-control (left), control-leptin (center), and cucurbitacin I-leptin (right). *, p < 0.05; scale bar = 100 μm.

Jacob C. Garza, et al. J Biol Chem. 2008 June 27;283(26):18238-18247.
6.
FIGURE 5.

FIGURE 5. From: Leptin Increases Adult Hippocampal Neurogenesis in Vivo and in Vitro.

Effects of leptin treatment on proliferation of adult hippocampal stem/progenitor cells. Cells were treated with various concentrations of leptin (1-30 nm) for 48 h and labeled with BrdUrd (10 μm) in the last 4 h of incubation. A, quantitative analysis revealed that the number of BrdUrd-labeled cells was increased by leptin treatment at concentrations of 1 nm and 3 nm when compared with control. Data are expressed as mean ± S.E. **, p < 0.01. B, representative microscopic images showing BrdUrd-labeled adult hippocampal progenitor cells in control (left panel) and leptin-treated groups (right panel). Scale bar = 100 μm.

Jacob C. Garza, et al. J Biol Chem. 2008 June 27;283(26):18238-18247.
7.
FIGURE 8.

FIGURE 8. From: Leptin Increases Adult Hippocampal Neurogenesis in Vivo and in Vitro.

Effect of inhibition of Akt on leptin-induced cell proliferation. Cultured adult hippocampal stem/progenitor cells were incubated with various concentrations of Akt inhibitor VIII (0.01-1.0 μm) for 4 h followed by treatment with leptin (1 nm) for 48 h and labeling with BrdUrd (10 μm) during the last 4 h of incubation. A, quantitative analysis revealed that pretreatment with Akt inhibitor VIII attenuates the leptin-induced increase in the number of BrdUrd-labeled cells in a dose-dependent manner. B, representative microscopic images of BrdUrd-labeled cells from three treatment conditions: control-control (left), control-leptin (center), and Akt inhibitor VIII-leptin (right). *, p < 0.05; scale bar = 100 μm.

Jacob C. Garza, et al. J Biol Chem. 2008 June 27;283(26):18238-18247.
8.
FIGURE 2.

FIGURE 2. From: Leptin Increases Adult Hippocampal Neurogenesis in Vivo and in Vitro.

Effect of leptin administration on cell differentiation. Mice were injected with leptin (1 mg/kg) or vehicle twice daily for 14 days followed by BrdUrd labeling and were perfused 28 days later. A, a significantly higher number of BrdUrd-positive cells remained in the leptin-treated group than in the control group 28 days after BrdUrd labeling. B, quantitative analysis showing the effect of leptin on the percentage of BrdUrd-labeled cells double labeled for NeuN or GFAP. Results are expressed as mean ± S.E. (n = 5 per group). *, p < 0.05, compared with the vehicle-treated control group. C, confocal microscopic images showing colocalization of BrdUrd with NeuN. D, confocal microscopic images showing colocalization of BrdUrd with GFAP. White arrows indicating double labeled cells. Scale bars in C and D = 50 μm for the images showing BrdUrd and NeuN/GFAP and 10 μm for the three-dimensional (3D) images.

Jacob C. Garza, et al. J Biol Chem. 2008 June 27;283(26):18238-18247.
9.
FIGURE 1.

FIGURE 1. From: Leptin Increases Adult Hippocampal Neurogenesis in Vivo and in Vitro.

Effect of leptin administration on cell proliferation in the dentate gyrus of adult mice. BrdUrd labeling was used to assess cell proliferation. Mice were injected intraperitoneally with leptin (1 mg/kg) or vehicle twice daily for 1, 5, or 14 days followed by BrdUrd labeling. The number of BrdUrd-labeled cells was counted in the subgranular zone and hilus of the dentate gyrus. A, acute treatment (1 day) with vehicle or leptin. B, short term treatment (5 days) with vehicle or leptin. C, chronic treatment (14 days) with vehicle or leptin. Images in D and E showing BrdUrd-labeled cells in the adult dentate gyrus of mice treated with vehicle (left panel) or leptin (right panel) for 14 days. Images in D′ and E′ showing BrdUrd-labeled cells in the paraventricular thalamus treated with vehicle (left panel) or leptin (right panel) for 14 days. The image in F indicates a high magnification view of proliferating cells. D3V, dorsal third ventricle; DG, dentate gyrus; GCL, granule cell layer; PVT, paraventricular thalamus; SGZ, subgranular zone. Scale bar = 100 μmin D and E, 50 μmin D′ and E′, and 10 μmin F. Data are expressed as the mean number of BrdUrd-labeled cells per dentate gyrus ± S.E. (n = 6 per group). **, p < 0.01.

Jacob C. Garza, et al. J Biol Chem. 2008 June 27;283(26):18238-18247.

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