Canine leptin: cDNA cloning, expression and activity of recombinant protein

Iwase, Miho; Kimura, Kazuhiro; Sasaki, Noriyuki; Komagome, Rika; Ishioka, Katsumi; Morimatsu, Masami; Murakami, Takashi; Saito, Masayuki

doi:10.1053/rvsc.1999.0342

Cited by 27 publications

(25 citation statements)

References 21 publications

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“…For example, a commercially available radioimmunoassay (RIA) kit for rat leptin shows 100% crossreactivity in rat, but less than 0.2% in human (manufacturer's instruction). In addition, we confirmed that anti-canine leptin antibody showed differences in immunoreactivity among other species as compared to the canine leptin [14]. Recently, Backus et al [1] reported serum leptin-like immunoreactiv-ity in cats was poorly measurable with a RIA based on antisera against mouse, rat, human, and primate leptins, suggesting the low immunoreactivity of these antisera to feline leptin.…”

Section: Discussionsupporting

confidence: 79%

“…The same missing sequence at the identical position was also reported in murine and canine leptins [14,19,27]. Since the CAG codon included a possible AG splice-acceptor sequence, there might have been slippage at this site.…”

Section: Discussionsupporting

confidence: 67%

“…Since the CAG codon included a possible AG splice-acceptor sequence, there might have been slippage at this site. Although Gln 49 is located in a highly conserved region of the leptin molecule, it has been suggested not to be essential for biological activities of leptin [14].…”

Section: Discussionmentioning

confidence: 99%

“…There have been several reports on complementary DNA (cDNA) cloning of leptin in various species [8,13,15,19,23], and we also reported cDNA cloning of canine leptin and the activity of its recombinant protein [14]. In this study, we cloned feline leptin cDNA using reverse transcription-polymerase chain reaction (RT-PCR) and examined tissue distribution of its mRNA by RT-PCR analysis.…”

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confidence: 98%

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cDNA Cloning of Feline Leptin and Its mRNA Expression in Adipose Tissue.

Sasaki

Shibata

Honjoh

et al. 2001

The Journal of Veterinary Medical Science

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ABSTRACT. Leptin, the product of the obese (ob) gene, is an adipocyte-derived hormone involved in regulating food intake and energy expenditure in humans and rodents. To determine the primary structure of feline leptin, we cloned the feline leptin cDNA using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of complementary DNA (cDNA) ends (RACE) methods. The full-length feline leptin cDNA was 2935 bp with a 501 bp open reading frame encoding the precursor peptide of 167 amino acids including 21 residues of signal peptide. The sequence of a 146-amino acid mature leptin was 81.5-91.8% homologous to those of other species. RT-PCR analysis revealed that the leptin mRNA was expressed in adipose tissues and not detected in liver, heart, kidney, lung, pancreas, brain and skeletal muscle. These data show that feline leptin is highly homologous to leptins of other species, and expressed in adipose tissues in cats. KEY WORDS: adipose tissue, cDNA cloning, feline, leptin.

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Section: Discussionsupporting

confidence: 79%

Section: Discussionsupporting

confidence: 67%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 98%

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cDNA Cloning of Feline Leptin and Its mRNA Expression in Adipose Tissue.

Sasaki

Shibata

Honjoh

et al. 2001

The Journal of Veterinary Medical Science

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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%

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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Seasonal changes in serum leptin of the feral raccoon (Procyon lotor) determined by canine-leptin-specific ELISA

et al. 2005

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Several reports have been published on blood leptin concentrations in feral animals, including members of the Carnivora, using a commercially available multi-species radioimmunoassay (RIA) kit with anti-human leptin antibody. However, we observed weak immunoreactivity between recombinant canine leptin and anti-human leptin antibody, suggesting a limitation in the applicability of the RIA kit for leptin assays in Carnivora species. We tested the applicability of RIA and sandwich enzyme-linked immunosorbent assay (ELISA) with anti-canine leptin antibody to assay blood leptin in the dog (Canis familiaris) and the raccoon (Procyon lotor). When RIA was used for recombinant canine leptin and dog sera, values were much lower than those determined by ELISA at higher concentrations (>10 ng/ml), while rather higher at lower concentrations (<2 ng/ml). A similar discrepancy between the two methods was found for serum leptin concentrations in raccoons. Clear seasonal variations were observed by ELISA, but not by RIA, with high values in autumn (3.46+/-0.45 ng/ml) and low values in spring and summer (0.71+/-0.07 ng/ml). Serum leptin concentrations in raccoons correlated positively with their body weight (r=0.753) and body mass index (r=0.755), corroborating our previous findings of a strong positive correlation between serum leptin concentrations and body fat content in dogs. Thus, the canine leptin ELISA is useful for assays of dog and raccoon leptin, and blood leptin is a good marker of nutritional condition in the species of Carnivora assayed in this study.

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Canine leptin: cDNA cloning, expression and activity of recombinant protein

Cited by 27 publications

References 21 publications

cDNA Cloning of Feline Leptin and Its mRNA Expression in Adipose Tissue.

cDNA Cloning of Feline Leptin and Its mRNA Expression in Adipose Tissue.

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

Seasonal changes in serum leptin of the feral raccoon (Procyon lotor) determined by canine-leptin-specific ELISA

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