Sandwich Enzyme-Linked Immunosorbent Assay of Canine Leptin.

Iwase, Miho; Kimura, Kazuhiro; Komagome, Rika; Sasaki, Noriyuki; Ishioka, K.; Honjoh, Tsutomu; Saito, Masayuki

doi:10.1292/jvms.62.207

Cited by 29 publications

(25 citation statements)

References 16 publications

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“…Although these low values may be a consequence of an inability of the kit to detect seal leptin rather than a reflection of true concentrations, circulating leptin concentrations in other carnivores are low (20). In seals, high assay variability combined with low actual leptin concentrations may mask important patterns of change in leptin secretion.…”

Section: Discussionmentioning

confidence: 99%

Molecular cloning and expression of leptin in gray and harbor seal blubber, bone marrow, and lung and its potential role in marine mammal respiratory physiology

Hammond

Bennett

Walton

et al. 2005

American Journal of Physiology-Regulatory, Integrative and Comp

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Section: Discussionmentioning

confidence: 99%

Molecular cloning and expression of leptin in gray and harbor seal blubber, bone marrow, and lung and its potential role in marine mammal respiratory physiology

Hammond

Bennett

Walton

et al. 2005

American Journal of Physiology-Regulatory, Integrative and Comp

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“…Since plasma leptin level is closely related to metabolic condition, it can be a strong tool for the study of obesity and related disorders. Previously, we cloned canine leptin gene (Iwase et al, 2000b), produced recombinant protein, and established a sandwich ELISA specific to canine leptin (Iwase et al, 2000a). Using this ELISA method, we demonstrated a highly positive correlation between plasma leptin concentration and body fat content estimated by a deuterium oxide dilution method in experimentally induced obese beagles (Sagawa et al, 2002, Ishioka et al, 2002b.…”

Section: Introductionmentioning

confidence: 93%

“…The plasma leptin concentration was measured by the previously reported method of sandwich ELISA using an anti-canine leptin antibody (Iwase et al, 2000a) with minor modifications. Briefly, each well of a 96-well microplate was coated with 0.2 µg of the purified rabbit anti-canine leptin antibody and incubated for 2 hr at room temperature.…”

Section: Leptin Assaymentioning

confidence: 99%

Plasma leptin concentration in dogs: Effects of body condition score, age, gender and breeds

Ishioka

Hosoya

Kitagawa

et al. 2007

Research in Veterinary Science

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Leptin is a cytokine produced by adipocytes, and plays a key role in the regulation of energy balance. In the present study, we measured plasma leptin concentrations of 166 normal and obese dogs visiting veterinary practices, and clarified the influence of age, gender and breed on plasma leptin levels in dogs. Leptin levels were higher in the dogs with higher body condition scores. There was no noticeable influence of age, gender and breed, but those in optimal puppies and obese Miniature Dachshund tended to be lower than those in corresponding groups. We conclude that plasma leptin is a reliable marker of adiposity in dogs regardless of age, gender and breed variations, and thereby useful as a blood biochemistry test for health examinations and treatment of obesity.2

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“…The plasma supernatant was transferred to 1.5-ml microfuge tubes and stored at Ϫ80°C until analyses were performed. Plasma leptin was measured with a sandwich ELISA for canine leptin as previously described (11). Briefly, rabbit anti-canine leptin antibodies were produced using canine recombinant leptin as an antigen.…”

mentioning

confidence: 99%

Leptin resistance extends to the coronary vasculature in prediabetic dogs and provides a protective adaptation against endothelial dysfunction

Knudson

Dinçer

Dick³

et al. 2005

American Journal of Physiology-Heart and Circulatory Physiology

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tinemia, associated with prediabetes, is an independent risk factor for coronary artery disease and a mediator of coronary endothelial dysfunction. We previously demonstrated that acutely raising the leptin concentration to levels comparable with those observed in human obesity significantly attenuates coronary dilation/relaxation to acetylcholine (ACh) both in vivo in anesthetized dogs and in vitro in isolated canine coronary rings. Accordingly, the purpose of this investigation was to extend these studies to a model of prediabetes with chronic hyperleptinemia. In the present investigation, experiments were conducted on control and high-fat-fed dogs. High-fat feeding caused a significant increase (131%) in plasma leptin concentration. Furthermore, in high-fat-fed dogs, exogenous leptin did not significantly alter vascular responses to ACh in vivo or in vitro. Coronary vasodilator responses to ACh (0.3-30.0 g/min) and sodium nitroprusside (1.0 -100.0 g/min) were not significantly different from those observed in control dogs. Also, high-fat feeding did not induce a switch to an endothelium-derived hyperpolarizing factor as a major mediator of muscarinic coronary vasodilation, because dilation to ACh was abolished by combined pretreatment with N -nitro-Larginine methyl ester (150 g/min ic) and indomethacin (10 mg/kg iv). Quantitative, real-time PCR revealed no significant difference in coronary artery leptin receptor gene expression between control and high-fat-fed dogs. In conclusion, high-fat feeding induces resistance to the coronary vascular effects of leptin, and this represents an early protective adaptation against endothelial dysfunction. The resistance is not due to altered endothelium-dependent or -independent coronary dilation, increased endothelium-derived hyperpolarizing factor, or changes in coronary leptin receptor mRNA levels. coronary circulation; metabolic syndrome; obesity HYPERLEPTINEMIA, universal in the human obese population (7), has recently been deemed an independent risk factor for cardiovascular disease (35) and, more specifically, a predictor of first myocardial infarction (43) and an independent risk factor for ischemic and hemorrhagic stroke (44). Since its discovery (53), leptin has been shown to promote platelet aggregation and thrombosis (3,16,17). Additionally, leptin is linked to the production of acute phase reactants (TNF-␣, IL-6, and IL-12) (22), neointimal growth in mice (38), superoxide production in aortic endothelium (50), and calcification of vascular smooth muscle cells (30). The leptin receptor is a type I cytokine receptor, similar to gp130 and granulocyte colony-stimulating factor receptors (45, 51), and elevated plasma leptin levels occur concurrently with elevated IL-6 and C-reactive protein in human obesity-related conditions (19,23,27). Despite the superfluity of studies linking leptin to various atherogenic processes, few studies to date have examined the direct effects of leptin on the coronary circulation.We (15) recently demonstrated that acutely raising coro...

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Sandwich Enzyme-Linked Immunosorbent Assay of Canine Leptin.

Cited by 29 publications

References 16 publications

Molecular cloning and expression of leptin in gray and harbor seal blubber, bone marrow, and lung and its potential role in marine mammal respiratory physiology

Molecular cloning and expression of leptin in gray and harbor seal blubber, bone marrow, and lung and its potential role in marine mammal respiratory physiology

Plasma leptin concentration in dogs: Effects of body condition score, age, gender and breeds

Leptin resistance extends to the coronary vasculature in prediabetic dogs and provides a protective adaptation against endothelial dysfunction

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