Now showing items 1-4 of 4

    • Article

      Differential expression of insulin-like growth factor I and II mRNAs during embryogenesis and early larval development in rabbitfish, Siganus guttatus 

      FG Ayson, EGT de Jesus, S Moriyama, S Hyodo, B Funkenstein, A Gertler & H Kawauchi - General and Comparative Endocrinology, 2002 - Academic Press
      In rodents, the expression of insulin-like growth factor II (IGF-II) is higher than that of insulin-like growth factor I (IGF-I) during fetal life while the reverse is true after birth. We wanted to examine whether this is also true in fish and whether IGF-I and IGF-II are differentially regulated during different stages of embryogenesis and early larval development in rabbitfish. We first cloned the cDNAs of rabbitfish IGF-I and IGF-II from the liver. Rabbitfish IGF-I has an open reading frame of 558 bp that codes for a signal peptide of 44 amino acids (aa), a mature protein of 68 aa, and a single form of E domain of 74 aa. Rabbitfish IGF-II, on the other hand, has an open reading frame of 645 bp that codes for a signal peptide of 47 aa, a mature protein of 70 aa, and an E domain of 98 aa. On the amino acid level, rabbitfish IGF-I shares 68% similarity with IGF-II. We then examined the relative expression of the two IGFs in unfertilized eggs, during different stages of embryogenesis, and in early larval stages of rabbitfish by a semiquantitative reverse transcription-polymerase chain reaction. Primers that amplify the mature peptide region of both IGFs were used and PCR for both peptides was done simultaneously, with identical PCR conditions for both. The identity of the PCR products was confirmed by direct sequencing. Contrary to published reports for seabream and rainbow trout, IGF-I mRNA was not detected in rabbitfish unfertilized eggs; it was first expressed in larvae soon after hatching. IGF-II mRNA, however, was expressed in unfertilized eggs, albeit weakly, and was already strongly expressed during the cleavage stage. mRNAs for both peptides were strongly expressed in the larvae, although IGF-II mRNA expression was higher than IGF-I expression.
    • Article

      Isolation, cDNA cloning, and growth promoting activity of rabbitfish (Siganus guttatus) growth hormone 

      FG Ayson, EGT de Jesus, Y Amemiya, S Moriyama, T Hirano & H Kawauchi - General and Comparative Endocrinology, 2000 - Elsevier
      We report the isolation, cDNA cloning, and growth promoting activity of rabbitfish (Siganus guttatus; Teleostei; Perciformes; Siganidae) growth hormone (GH). Rabbitfish GH was extracted from pituitary glands under alkaline conditions, fractionated by gel filtration chromatography on Sephadex G-100, and purified by high-performance liquid chromatography. The fractions containing GH were identified by immunoblotting with bonito GH antiserum. Under nonreducing conditions, the molecular weight of rabbitfish GH is about 19 kDa as estimated by SDS–PAGE. The purified hormone was potent in promoting growth in rabbitfish fry. Weekly intraperitoneal injections of the hormone significantly accelerated growth. This was evident 3 weeks after the start of the treatment, and its effect was still significant 2 weeks after the treatment was terminated. Rabbitfish GH cDNA was cloned to determine its nucleotide sequence. Excluding the poly (A) tail, rabbitfish GH cDNA is 860 base pairs (bp) long. It contained untranslated regions of 94 and 175 bp in the 5′ and 3′ ends, respectively. It has an open reading frame of 588 bp coding for a signal peptide of 18 amino acids and a mature protein of 178 amino acid residues. Rabbitfish GH has 4 cysteine residues. On the amino acid level, rabbitfish GH shows high identity (71–74%) with GHs of other perciforms, such as tuna, sea bass, yellow tail, bonito, and tilapia, and less (47–49%) identity with salmonid and carp GHs.
    • Article

      Milkfish (Chanos chanos) growth hormone cDNA cloning and mRNA expression in embryos and early larval stages. 

      EGT de Jesus, FG Ayson, Y Amemiya, S Moriyama, S Hyodo, T Hirano & H Kawauchi - Aquaculture, 2002 - Elsevier
      In an attempt to understand growth regulation in milkfish, the milkfish growth hormone (GH) and its cDNA were characterized and the expression of GH mRNA in embryos and larvae was examined by RT-PCR. The milkfish GH was purified from an alkaline extract of the pituitary by reverse-phase high-performance liquid chromatography and detected as an immuno-positive protein with anti-salmon GH serum. The complete sequence of milkfish pre-GH was determined by cDNA cloning and nucleotide sequencing. On the basis of the N-terminal amino acid analysis of the native protein, the pre-GH was found to consist of a signal peptide of 22 amino acids and a mature protein of 188 amino acids. Milkfish GH shows higher amino acid sequence identity with GHs of carps (91–94%) and salmonids (70%) than with GHs of more advanced teleosts (<60%) in good accordance with its taxonomic position in teleosts. It has five half Cys residues, four of which are at positions homologous with those of other known GHs and the extra Cys with those of carp GHs. The molecular weight of milkfish GH was estimated to be 22 kDa, which is comparable to the theoretical value. This suggests that milkfish GH is a simple protein, although it has two potential N-glycosylation sites. Semiquantitative RT-PCR showed that GH mRNA expression was relatively weak in embryos and newly hatched larvae but was already strong in 2-day old and older larvae.
    • Article

      Thyroid hormones promote early metamorphosis in grouper (Epinephelus coioides) larvae 

      EGT de Jesus, JD Toledo & MS Simpas - General and Comparative Endocrinology, 1998 - Academic Press
      The response of grouper larvae to the thyroid hormones, thyroxine (T4) and triiodothyronine (T3), was examined. Two-, 3-, and 4-week-old grouper larvae were reared in seawater containing either T4 or T3 at 0.01, 0.1, and 1 ppm. T4 or T3 induced metamorphosis in all age groups in a dose-dependent manner. Regardless of the size of the larvae, metamorphosis was completed in 2 days in larvae treated with 1 ppm of either T4 or T3; 3–4 days in larvae exposed to 0.1 ppm; and 5–6 days in larvae immersed in 0.01 ppm. None of the fish in the control group completed metamorphosis during this period. Compared with the control fish, survival rates were higher in groups exposed to 0.01 ppm and lower in those exposed to 1 ppm of T3. In 4-week-old larvae, T4 treatment (0.01 to 1.0 ppm) resulted in higher survival compared to the control. These results suggest that a dose of 0.01 ppm is appropriate for acceleration of metamorphosis and improvement of survival in 3- and 4-week-old grouper larvae. A lower dose may be apropriate for earlier stages.