ABSTRACT
The full-length of insulin-like growth factor (IGF) complementary (c)DNAs encoded by igf-I and igf-II from torafugu pufferfish Takifugu rubripes were cloned in the present study. The deduced amino acid sequences of the two genes showed c. 80% identity each with those of Igf-I and Igf-II from other teleosts, respectively. Two growth hormone (GH) receptors, ghr1 and ghr2, were also cloned in silico using the T. rubripes Fugu genome database. The transcripts of T. rubripes igf-I were detected in slow muscle, heart, skin, gill, liver and intestine but not in fast muscle, spleen and testis of adult fish, whereas those of igf-II were found in all tissues examined. Subsequently, the accumulated messenger (m)RNA levels of igf-I and igf-II were investigated in an F(2) population derived from a male of an apparent fast-growing T. rubripes strain and a wild female T. rubripes together with those of other growth-related genes encoding Gh, Ghr1 and Ghr2, and with those of prolactin (Prl) and leptin (Lep) previously reported. The accumulated mRNA levels of igf-I, gh and ghr1 were significantly correlated to growth rate at larval stages in the population, but not for those of igf-II, prl, ghr2 and lep. Although it is unclear whether or not this phenotype is directly related to the heredity of the fast-growing strain, the findings suggest that the expression of igf-I, gh and ghr1 is involved in the regulation of growth rate at larval stages in T. rubripes.
Subject(s)
Body Size , Gene Expression Regulation , Growth Hormone/genetics , Insulin-Like Growth Factor I/genetics , RNA, Messenger/metabolism , Receptors, Somatotropin/genetics , Animals , Takifugu/anatomy & histology , Takifugu/growth & developmentSubject(s)
Antineoplastic Agents/blood , Nitrosourea Compounds/blood , Streptozocin/analogs & derivatives , Antineoplastic Agents/urine , Chromatography, High Pressure Liquid , Humans , Nitrosourea Compounds/pharmacology , Nitrosourea Compounds/urine , Streptozocin/blood , Streptozocin/pharmacology , Streptozocin/urine , Time FactorsABSTRACT
A high-performance liquid chromatographic (HPLC) method for the quantitative determination of 1-hexylcarbamoyl-5-fluorouracil (HCFU) and its metabolites using mu Bondapak C18 and mu Porasil has been developed. Two mobile phases containing PIC-B7 (consisting of acetic acid and 1-heptanesulphonic acid) were used for the separation, and good separations were obtained. With methanol-water (56:44) as the mobile phase, the separation of HCFU and its three metabolites was achieved within 4 min. With methanol-water (32:68) a new metabolite, 1-omega-carboxymethylcarbamoyl-5-fluorouracil, was revealed in human plasma. The recovery of each substance was 80% or greater and the sensitivity was at the nanograms per millilitre level. The coefficient of variation was less than 3.6% for each component.
