Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Central overexpression of leptin antagonist reduces wheel running and underscores importance of endogenous leptin receptor activity in energy homeostasis

doi: 10.1152/ajpregu.90449.2008

Figure Lengend Snippet: Leptin antagonist overexpression in hypothalamus. Leptin antagonist transgene expression (lower band), determined by using RT-PCR, was evident in all rats receiving recombinant adeno-associated virus (rAAV)-leptin antagonists (A) and absent in those receiving control vector (C). Upper band represents 18S ribosomal RNA.

Article Snippet: Production of rAAV Vectors Rat leptin antagonist DNA was constructed by PCR-based mutagenesis of the rat leptin gene (SeqWright, Houston, TX) to yield the sequence of a protein previously characterized to block leptin receptor signaling and leptin-induced physiological responses ( 35 ).

Techniques: Over Expression, Expressing, Reverse Transcription Polymerase Chain Reaction, Recombinant, Virus, Control, Plasmid Preparation

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Central overexpression of leptin antagonist reduces wheel running and underscores importance of endogenous leptin receptor activity in energy homeostasis

doi: 10.1152/ajpregu.90449.2008

Figure Lengend Snippet: Top: body weight in rats after administration of control vector or high-dose rAAV antagonist. rAAV antagonist or control vectors were administered at day 0 in rats maintained on a chow diet for 82 days. After a series of tests and blood draw, the rats were switched to a 60% high-fat (HF) diet until day 140, after which they were returned to the chow diet for the remainder of the experiment. WR indicates days on which wheel running was evaluated. Values represent means ± SE of 5 rats/group. Bottom: change (Δ) in body weight over the indicated days, representing the initial chow feeding period after recovery from surgery and before blood draw (left), the HF feeding period before evaluation of WR (center), and the final chow feeding period before evaluation of WR (right). *P = 0.0005, difference from control by t-test.

Article Snippet: Production of rAAV Vectors Rat leptin antagonist DNA was constructed by PCR-based mutagenesis of the rat leptin gene (SeqWright, Houston, TX) to yield the sequence of a protein previously characterized to block leptin receptor signaling and leptin-induced physiological responses ( 35 ).

Techniques: Control, Plasmid Preparation

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Central overexpression of leptin antagonist reduces wheel running and underscores importance of endogenous leptin receptor activity in energy homeostasis

doi: 10.1152/ajpregu.90449.2008

Figure Lengend Snippet: Top: daily caloric consumption after administration of control vector or high-dose rAAV antagonist during the period of chow feeding. Inset: cumulative food consumption is significantly elevated between days 12 and 72. *P = 0.016, difference in cumulative food consumption during this period. Bottom: caloric consumption after initiation of HF feeding at day 83 after administration of control vector or rAAV antagonist. Inset: cumulative food consumption is significantly elevated between days 84 and 93. Values represent means ± SE of 5 rats/group. *P = 0.019 for difference in cumulative food consumption.

Article Snippet: Production of rAAV Vectors Rat leptin antagonist DNA was constructed by PCR-based mutagenesis of the rat leptin gene (SeqWright, Houston, TX) to yield the sequence of a protein previously characterized to block leptin receptor signaling and leptin-induced physiological responses ( 35 ).

Techniques: Control, Plasmid Preparation

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Central overexpression of leptin antagonist reduces wheel running and underscores importance of endogenous leptin receptor activity in energy homeostasis

doi: 10.1152/ajpregu.90449.2008

Figure Lengend Snippet: Feed efficiency (body weight gain per unit of caloric consumption) with chow and HF feeding

Article Snippet: Production of rAAV Vectors Rat leptin antagonist DNA was constructed by PCR-based mutagenesis of the rat leptin gene (SeqWright, Houston, TX) to yield the sequence of a protein previously characterized to block leptin receptor signaling and leptin-induced physiological responses ( 35 ).

Techniques:

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Central overexpression of leptin antagonist reduces wheel running and underscores importance of endogenous leptin receptor activity in energy homeostasis

doi: 10.1152/ajpregu.90449.2008

Figure Lengend Snippet: Change in body weight in rats after administration of control vector or the lower dose of rAAV antagonist. rAAV antagonist or control vectors were administered at day 0 in rats maintained on a chow diet for 87 days. Body weight was significantly different between groups commencing at day 27.

Article Snippet: Production of rAAV Vectors Rat leptin antagonist DNA was constructed by PCR-based mutagenesis of the rat leptin gene (SeqWright, Houston, TX) to yield the sequence of a protein previously characterized to block leptin receptor signaling and leptin-induced physiological responses ( 35 ).

Techniques: Control, Plasmid Preparation

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Central overexpression of leptin antagonist reduces wheel running and underscores importance of endogenous leptin receptor activity in energy homeostasis

doi: 10.1152/ajpregu.90449.2008

Figure Lengend Snippet: Top: WR activity during chow and HF feeding periods in rats after administration of control vector or rAAV-leptin antagonist. WR was assessed for a 4-day period during chow feeding after the low dose of the antagonist vector (days 34–38) and during HF feeding (days 130–134) and chow feeding (days 184–188) after the high dose of antagonist vector. Values represent means ± SE of 5 or 6 rats/group. *P = 0.001, difference with antagonist treatment; **P = 0.03, difference between control chow-fed and control HF-fed rats after high dose of antagonist vector. Bottom: WR activity after administration of control vector, rAAV-leptin, or control vector pair fed the amount of food consumed by the rAAV-leptin group. WR was assessed for a 4-day period starting at day 105 after vector delivery. Values represent means ± SE of 8–10 rats/group. *P = 0.001 for difference with leptin treatment from control or pair fed by 1-way ANOVA.

Article Snippet: Production of rAAV Vectors Rat leptin antagonist DNA was constructed by PCR-based mutagenesis of the rat leptin gene (SeqWright, Houston, TX) to yield the sequence of a protein previously characterized to block leptin receptor signaling and leptin-induced physiological responses ( 35 ).

Techniques: Activity Assay, Control, Plasmid Preparation

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Central overexpression of leptin antagonist reduces wheel running and underscores importance of endogenous leptin receptor activity in energy homeostasis

doi: 10.1152/ajpregu.90449.2008

Figure Lengend Snippet: Top: whole body adiposity (left) and % adiposity relative to body weight (right) before vector delivery (day 0), before HF feeding (day 80), during HF feeding (days 106, 127, and 138), and after return to a chow diet (day 183) in control and high-dose rAAV antagonist-treated rats. Values represent means ± SE of 5 rats/group. P < 0.01 for difference between control and rAAV antagonist at each assessment day except day 0. Bottom: serum leptin before vector delivery (day 0), before HF feeding (day 77), during HF feeding (day 106), and after return to a chow diet (day 183) in control and high-dose rAAV antagonist-treated rats. Values represent means ± SE of 5 rats/group. P < 0.001 (day 77) and P < 0.05 (day 106) for difference between control and rAAV antagonist.

Article Snippet: Production of rAAV Vectors Rat leptin antagonist DNA was constructed by PCR-based mutagenesis of the rat leptin gene (SeqWright, Houston, TX) to yield the sequence of a protein previously characterized to block leptin receptor signaling and leptin-induced physiological responses ( 35 ).

Techniques: Plasmid Preparation, Control

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Central overexpression of leptin antagonist reduces wheel running and underscores importance of endogenous leptin receptor activity in energy homeostasis

doi: 10.1152/ajpregu.90449.2008

Figure Lengend Snippet: Rat weight, adiposity, and UCP1 protein levels at death

Article Snippet: Production of rAAV Vectors Rat leptin antagonist DNA was constructed by PCR-based mutagenesis of the rat leptin gene (SeqWright, Houston, TX) to yield the sequence of a protein previously characterized to block leptin receptor signaling and leptin-induced physiological responses ( 35 ).

Techniques:

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Central overexpression of leptin antagonist reduces wheel running and underscores importance of endogenous leptin receptor activity in energy homeostasis

doi: 10.1152/ajpregu.90449.2008

Figure Lengend Snippet: Signal transducer and activator of transcription 3 (STAT3) phosphorylation following a single intracerebroventricular injection of either a submaximal (40 ng) or a supramaximal (250 ng) of leptin in control vector- or low-dose rAAV-leptin antagonist-treated rats. STAT3 phosphorylation was assessed at day 87 before termination of the experiment. Values represent % increase over vehicle-injected rats and are means ± SE of 6 rats/group. *P = 0.034, difference from control vector by t-test.

Article Snippet: Production of rAAV Vectors Rat leptin antagonist DNA was constructed by PCR-based mutagenesis of the rat leptin gene (SeqWright, Houston, TX) to yield the sequence of a protein previously characterized to block leptin receptor signaling and leptin-induced physiological responses ( 35 ).

Techniques: Phospho-proteomics, Injection, Control, Plasmid Preparation

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Central overexpression of leptin antagonist reduces wheel running and underscores importance of endogenous leptin receptor activity in energy homeostasis

doi: 10.1152/ajpregu.90449.2008

Figure Lengend Snippet: Hypothalamic signaling factors

Article Snippet: Production of rAAV Vectors Rat leptin antagonist DNA was constructed by PCR-based mutagenesis of the rat leptin gene (SeqWright, Houston, TX) to yield the sequence of a protein previously characterized to block leptin receptor signaling and leptin-induced physiological responses ( 35 ).

Techniques: