[Show abstract][Hide abstract] ABSTRACT: We have cloned two cDNAs from the pituitary gland of blue gourami (Trichogaster trichopterus), coding for the beta subunits of the gonadotropin hormones GtH-I and GtH-II. The two cDNAs were sequenced and subjected to sequence analysis. We have found that the deduced amino acid sequences of both cDNAs were most similar to their striped bass counterparts. The beta GtH-I subunits of blue gourami and striped bass shared 73% of their residues, and the beta GtH-II subunits 84%. The cloning of the cDNAs of beta GtH-I and beta GtH-II has enabled us to measure the expression of their respective mRNAs in the pituitaries of female blue gourami at different stages of the reproductive cycle. The highest levels of beta GtH-I and beta GtH-II mRNA were found in specimens classified as high vitellogenic and in females that were at the final stages of oocyte maturation.
Journal of Molecular Endocrinology 11/1999; 23(2):177-87. · 3.08 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Profiles of plasma growth hormone (GH) in male tilapia hybrid (Oreochromis niloticus x O. aureus) were measured and compared at different times of the year. The profiles did not appear to be repetitive, however, differences in their nature were observed at the different seasons; the most erratic profiles were seen in the height of the reproductive season (July), while the peaks were more subdued in the spring and disappeared in the autumn. Peaks in male fish were more prominent than in the females when measured in July. Perifused pituitary fragments from fish with a high GSI responded to salmon gonadotropin-releasing hormone (sGnRH) analog (10 nM-1 μM), while those from fish with a low GSI barely responded to even the highest dose. Exposure of perifused pituitary fragments from sexually-regressed fish to carp growth hormone-releasing hormone (cGHRH; 0.1 μM) or sGnRH (I μM) stimulated GH release only after injection of the fish with methyl testosterone (MT; 3 injections of 0.4 mg kg (1)). The same MT pretreatment did not alter the response to dopamine (DA; 1 or 10 μM). GH pituitary content in MT-treated fish was lower than in control fish, which may be explained by the higher circulating GH levels in these fish, but does not account for the increased response to the releasing hormones. Castration abolished the response of cultured pituitary cells to sGnRH (I fM-100 nM) without altering either their basal rate of secretion or circulating GH levels. Addition of steroids to the culture medium (MT or estradiol at 10 nM for 2 days) enabled a GH response to sGnRH stimulation in cells from sexually regressed fish. Pituitary cells which had not been exposed to steroids failed to respond to sGnRH, although their response to forskolin or TPA was similar to that of steroid-exposed cells. It would appear, therefore, that at least one of the effects of the sex steroids on the response to GnRH is exerted proximally to the formation of cAMP, or PKC, presumably at the level of the receptor. An increase in the number of receptors to the GH-releasing hormones, following steroid exposure, would explain also the changing nature of the GH secretory profile in different stages of the reproductive season.
Fish Physiology and Biochemistry 08/1995; 14(4):267-77. DOI:10.1007/BF00004065 · 1.62 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The present study is an attempt to find sites of dopaminergic inhibition along the transduction cascades culminating in gonadotropin (GtH) release in a teleost fish, tilapia. Experiments were carried out on perifused pituitary fragments and in primary culture of trypsinized pituitary cells. Salmon GnRH, chicken GnRH I and II stimulated GtH release in culture with estimated ED50 values of 15.56 pM, 2.55 nM and 8.65 pM, respectively. Apomorphine (APO; 1 microM) totally abolished this stimulation. Dopamine (DA; 1 microM) reduced both basal and GnRHa-stimulated GtH release from perifused pituitary fragments but did not alter the formation of cAMP. In a similar perifusion experiment DA abolished GtH release in response to forskolin (10 microM) with no reduction in cAMP formation. This indicates that one site of the dopaminergic inhibition is distal to cAMP formation, an indication not compatible with the classic characteristic of DA D2 type mode of action. The inhibition of GtH release in culture, caused by 1 microM APO, the specific DA D2 agonists LY 171555 (LY) or bromocryptine (BRCR) could not be reversed by activating protein kinase C (PKC) by DiC8 or the phorbol ester TPA. This would indicate a site for DA action distal to PKC. However, the stimulatory effect of arachidonic acid (AA; 50 microM) in perifusion was not reduced by DA (1 microM) or by APO, LY or BRCR in culture, which suggests a site for DA action proximal to AA formation. APO, LY and BRCR reduced GtH release in response to the Ca2+ ionophore A23187, however, their inhibitory effect was reversed by 10 microM ionomycin.(ABSTRACT TRUNCATED AT 250 WORDS)
[Show abstract][Hide abstract] ABSTRACT: A radioimmunoassay (RIA) for recombinant tilapia growth hormone (GH) was established and validated. The ability of various hypothalamic factors to regulate GH secretion in the tilapia hybrid (Oreochromis niloticus × Oreochromis aureus) was studied. Somatostatin1-14 (SRIF1-14; 10-100 μg/kg) was found to reduce circulating GH levels in a dose-dependent manner. SRIF1-14 (0.1-1000 nM) inhibited GH release from perifused pituitary fragments (ED50 0.83 nM). Human growth hormone-releasing hormone fragment 1-29 (hGHRH1-29; 100 μg/kg) doubled circulating GH levels and modestly stimulated GH secretion in vitro. Carp growth hormone-releasing hormone (cGHRH) stimulated GH secretion in vitro to a similar degree at the same dose (1 μM). Injection of salmon gonadotropin-releasing hormone (sGnRH) superactive analog (10-100 μg/kg) increased plasma GH levels sixfold. sGnRH also stimulated GH release in vitro (ED50 142.56 nM). Dopamine (0.1-10 μM) and the D1 DA receptor agonist SKF 38393 increased GH secretion from perifused pituitary fragments dose-relatedly. Thyrotropin-releasing hormone (TRH) had no effect on GH secretion from perifused pituitary fragments, but increased plasma GH levels, as did bovine thyroid stimulating hormone (bTSH). The increased plasma GH in the bTSH-treated fish coincided with a dramatic increase in T4; however, TRH increased GH without changing T4 levels. T3 increased the synthesis of GH by isolated pituitaries (incorporation of [3H]leucine). SRIF1-14 seems to be a most potent hypothalamic regulator of GH secretion in tilapia; sGnRH and DA both increased GH secretion, although sGnRH elicited considerably greater responses at lower doses. Two forms of GHRH increased GH levels, although the unavailability of the homologous peptide prevented an accurate evaluation of its importance in regulating GH secretion. The thyroid axis (TRH, TSH, and T3) stimulates both synthesis and release of GH, although TRH did not appear to have a direct effect on the level of the pituitary.
General and Comparative Endocrinology 02/1995; 97(1-97):13-30. DOI:10.1006/gcen.1995.1002 · 2.47 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Triploidy as a result of thermal shock exposure of fertilized eggs decreases the growth rate ofOreochromis aureus as compared to their diploid controls, but this is due to the higher female ratio present in triploids (86%) and the lower growth rate of females. When females and males are considered separately, the growth rate is not significantly different in diploids and triploids. Since triploidy results in a malfunctioning steroidogenesis in females (mainly testosterone (T) and 17β-estradiol (E2)), but does not affect the growth rate, it is concluded that female gonadal steroids do not influence growth unless in pharmacological concentrations. These low levels of gonadal steroids are generally accompanied by higher levels of gonadotropin (GtH), but the difference is not always significant.Despite their lower growth rate diploid females have higher plasma concentrations of growth hormone (GH) during several months compared to the triploid females and diploid males. 3,5,3'-triiodo-L-thyronine (T3) levels, however, are comparable between diploid and triploid females (except for 1 month), but higher in diploid males in 4 of the 5 months studied. 11-ketotestosterone (11kT) is always higher in males. These results indicate that the higher growth rate of males may be related to the high circulating levels of T3 and 11kT.
Fish Physiology and Biochemistry 07/1994; 13(3):209-18. DOI:10.1007/BF00004359 · 1.62 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Spawning experiments under routine hatchery conditions on Israeli farms showed that successful ovulation in the common carp (Dor 70×Yugoslavian lines) can be achieved by a single administration of 10 μg/kg sGnRHa ([D-Arg6,Pro9-NEt]-sGnRH) combined with 20 mg/kg of the water-soluble dopamine receptor antagonist, metoclopramide (GnRH + MET). An initial rise in the maturational gonadotropin (cGtH) level occurred 3 h after treatment, gradually increasing thereafter, and reaching a peak of 227±41.8 ng/ml (mean ± s.e.m., n = 10) 14 h after treatment, when full ovulation took place, as reflected by the presence of a few expelled eggs on the bottom of the tank. This rise was associated with increased levels of oestradiol-17β (E2; 19.5 ± 3.4 ng/ml) and 17α,20β-dihydroxy-4-pregnen-3-one (17,20-P; 23.9±19.7 ng/ml). Ovarian biopsies showed a gradual progress in oocyte maturation with germinal vesicle breakdown (GVBD) occurring during the rise in both steroids. At the time of initial egg release, GtH remained at a high level while the steroids started to decline. In a parallel group, fish were primed with a dose of carp pituitary extract (CPE), calibrated to contain 0.07 mg/kg cGtH, and 11 h later with a resolving dose of CPE (0.34 mg/kg cGtH). Following priming, circulating GtH and E2 increased moderately, and the germinal vesicle migrated towards the oocytes' periphery. A further increase in cGtH and E2, and a sharp peak in 17,20-P, occurred concomitantly with GVBD, 6 h after the resolving dose was given. Data compiled from several spawning experiments showed that the interval between hormone administration and initial egg release (latency) was negatively correlated with water temperature over the range of 20–26°C. While latency was always longer in GnRH+MET than in CPE treatment, in each treatment it was nonetheless constant between 22.5°C and 25°C. GnRH+MET given at various hours of the day or evening resulted in identical spawning with the same latency. This fact, together with the predicted latency at a given temperature, may provide the aquaculturist with a protocol to accurately schedule spawning induction in the common carp.