Determination of rat leptin activity in vitro using a novel luciferase reporter assay.
Leptin is small cytokine-like protein that is involved in appetite and body weight regulation. Due to increased interest in using leptin as an anti-obesity reagent, recombinant forms of leptin have been produced for several species, including humans, mice, rats, pigs, dogs, sheep etc. The biological activities of such recombinant proteins were determined using various in vitro or in vivo systems; however so far, no specific assay system for rat leptin is available. Since rats are representative animal models in obesity research, the establishment of a biological assay system for determining rat leptin activity has been eagerly awaited. This study describes the generation of such a system using chinese hamster ovary (CHO)-cells that were transfected with the long form of the rat leptin receptor isoform, OB-Rb, whereby a signal transduces and activators of transcription-sensitive luciferase reporter system is further employed to quantify the leptin-mediated signals. This system is the first rat-specific leptin bioassay system that has been reported. It is expected that this assay will be used to further quantify and determine leptin activity from various biological fluids and sources.
Ju
S K
SK
Protein Engineering Laboratory, Korea Research Institute of Bioscience and Biotechnology, Taejon.
Park
J H
JH
Na
S Y
SY
You
K H
KH
Kim
K L
KL
Lee
M K
MK
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D016466:CHO Cells; D002352:Carrier Proteins / Q000378:metabolism*; D006224:Cricetinae; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D020738:Leptin / Q000378:metabolism*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D051381:Rats; D011956:Receptors, Cell Surface; D054411:Receptors, Leptin; D011994:Recombinant Proteins / Q000378:metabolism
D002352:Carrier Proteins; C515448:LEPR protein, human; D020738:Leptin; D011956:Receptors, Cell Surface; D054411:Receptors, Leptin; D011994:Recombinant Proteins; D008156:Luciferases