Effects of growth hormone over-expression on reproduction in the common carp Cyprinus carpio L.

Growth hormone (GH) transgenic common carp (Cyprinus carpio L.) show desirable aquaculture traits. Their specific growth rate (SGR) and feed efficiency (FE) are approximately 12% and 17% higher than the wild-type (WT) common carp, respectively. However, the mechanisms of lipid catabolism (lipolysis and fatty acid β-oxidation) and utilization in GH transgenic common carp are still unclear. In this study, we firstly compared the lipid metabolism of GH transgenic (initial weight 3.72 ± 0.32 g) and WT (initial weight 3.30 ± 0.28 g) common carp fed with a normal fat level diet (6% lipid, 33% protein) for two months, then compared the growth performance of GH transgenic (initial weight 3.65 ± 0.33 g) and WT (initial weight 3.27 ± 0.26 g) common carp fed with different fat levels diets (6% lipid and 12% lipid, 33% protein) for two months. We found that the lipid content in serum, liver and whole body was significantly reduced in GH transgenic common carp, the hepatic activities of the lipolytic enzymes hormone-sensitive lipase and adipose triglyceride lipase were enhanced, and the hepatic expression level of hormone-sensitive lipase was upregulated. In addition, the mitochondrion numbers were increased, and the expression level of carnitine palmitoyltransferase-1a and carnitine palmitoyltransferase-1b was upregulated in the liver of GH transgenic common carp. GH transgenic common carp showed higher weight gain and SGR than that in WT carp when fed with a normal-fat diet as they did when fed with a high-fat diet, and GH transgenic common carp showed higher FE than that in WT carp when fed with a high-fat diet. These results suggested that the lipid catabolism and utilization was improved in the GH transgenic common carp liver through enhanced lipolytic and fatty acid β-oxidation pathways. Our study provides new insights into improving lipid utilization in some aquaculture fish species.

The hematological response to exhaustive exercise in ‘all-fish’ growth hormone transgenic common carp (Cyprinus carpio L.)

Increased food intake in growth hormone-transgenic common carp (Cyprinus carpio L.) may be mediated by upregulating Agouti-related protein (AgRP)

The survival and tolerance of F2 transgenic common carp (Cyprinus carpio) containing pRSVrtGH1 complementary DNA were compared with nontransgenic (control) common carp when subjected to low dissolved oxygen. The tolerance of low oxygen was evaluated in 8 families of common carp in rectangular tanks (3 x 1 x 1 m). The absolute mean percentage of survival of transgenic common carp subjected to low oxygen (0.4 mg/L) was higher (P <0.05) than that of control carp in 2 of the 8 families of common carp tested; however, the overall means for all families of transgenic and control carp were not different (P > 0.05). When oxygen tolerance was measured in time to death rather than absolute survival or mortality, the growth hormone transgenic common carp had a longer group mean (P <0.05) than did controls. The mean survival time in minutes for the transgenic genotype was greater (P <0.05) in 5 of the 8 families assessed. Transgenic common carp in some families had higher percentage and longer times of survival than control common carp when subjected to low oxygen. The definition of tolerance of low oxygen and how it is measured is important, and can affect interpretation of results. The pleiotropic effect of pRSVrtGH1 cDNA on superior survival of low oxygen in common carp has important implications for intensive fish culture.

Metabolism traits of ‘all-fish’ growth hormone transgenic common carp ( Cyprinus carpio L.)

Acute and chronic un-ionized ammonia toxicity to ‘all-fish’ growth hormone transgenic common carp (Cyprinus carpio L.)

Effects of gonadotropin-releasing hormone (GnRH) analogs and sex steroids on growth hormone (GH) secretion and growth in common carp ( Cyprinus carpio) and grass carp ( Ctenopharyngodon idellus)

The Influence of Pharmacological Manipulation of Serotonin on Serum Growth Hormone and Luteinizing Hormone Levels in Young Turkeys (Meleagris gallopavo)

Studies on the luteinizing hormone- and follicle-stimulating hormone-releasing mechanism in the testicular feminization syndrome: Hypothalamic-pituitary responsiveness to clomiphene, luteinizing hormone-releasing hormone, gonadectomy, and sexual steroids

Food consumption, number of movements and feeding hierarchy of juvenile transgenic common carp Cyprinus carpio and their size-matched non-transgenic conspecifics were measured under conditions of limited food supply. Transgenic fish exhibited 73.3% more movements as well as a higher feeding order, and consumed 1.86 times as many food pellets as their non-transgenic counterparts. After the 10 day experiment, transgenic C. carpio had still not realized their higher growth potential, which may be partly explained by the higher frequency of movements of transgenics and the 'sneaky' feeding strategy used by the non-transgenics. The results indicate that these transgenic fish possess an elevated ability to compete for limited food resources, which could be advantageous after an escape into the wild. It may be that other factors in the natural environment (i.e. predation risk and food distribution), however, would offset this advantage. Thus, these results need to be assessed with caution.

Excitatory amino acids, such as glutamate, constitute a major transmitter system in the control of hypothalamic-pituitary secretion. Different subtypes of glutamate receptors, such as NMDA (N-methyl-d-aspartic acid) and KA (kainate) receptors, are involved in the control of anterior pituitary secretion. Other receptor subtypes, such as AMPA (activated by alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid) and metabotropic receptors, have been identified, although their role in the control of neuroendocrine function remains largely unknown. Recent reports have demonstrated the involvement of AMPA receptors in the control of the steroid-induced luteinizing hormone (LH) surge in female and growth hormone (GH) secretion in male rats. The aim of this study was to assess the potential role of AMPA receptors in the control of GH, prolactin (PRL), LH and follicle-stimulating hormone (FSH) secretion in prepubertal 23-day-old rats. To this end, prepubertal female rats were injected with AMPA (2.5 or 5 mg/kg i.p.) or the antagonist of AMPA receptors 1,2,3,4-tetrahydro-6-nitro-2, 3-dioxo-benzo (f) quinoxaline-7-sulfonamide (NBQX; 0.25 or 0.50 mg/kg i.p.). Serum LH and FSH concentrations and hypothalamic LH-releasing hormone (LHRH) content remained unchanged after AMPA or NBQX administration. In contrast, serum PRL levels significantly decreased 15, 30 and 60 min after i.p. administration of AMPA and increased 120 min after NBQX treatment, whereas serum GH levels increased after AMPA treatment and decreased after NBQX administration. Considering that AMPA has been shown to activate a subset of kainate receptors, its effects were compared with those elicited by 2.5 mg/kg KA in prepubertal female rats. At this age, however, KA was unable to reproduce the effects of AMPA on PRL and GH secretion, thus suggesting that the actions observed after AMPA administration were carried out specifically through AMPA receptors. In addition, as the effects of AMPA on LH secretion in adult females have been proved to be steroid-dependent, the effects of AMPA (2.5 mg/kg) and NBQX (0.5 mg/kg) were tested in prepubertal animals with different gonadal backgrounds, i.e. intact males, and intact and ovariectomized (OVX) females. The effects of AMPA in prepubertal females appeared to be modulated by ovarian secretion, as the inhibition of PRL secretion disappeared and LH secretion was partially suppressed by AMPA in OVX animals whereas the stimulatory effect on GH release was enhanced by ovariectomy. Furthermore, in male rats, AMPA administration significantly decreased PRL secretion and increased serum GH levels, the amplitude of the GH response being higher than in prepubertal females. To ascertain the pituitary component for the reported actions of AMPA, hemi-pituitaries of male rats were incubated in the presence of AMPA (10(-8)-10(-6) M). The results obtained showed no effect of AMPA on PRL, GH and gonadotropin secretion in vitro. Finally, we investigated the involvement of the dopaminergic (DA) system in the inhibitory action of AMPA on PRL secretion. Pre-treatment of prepubertal female rats with a dopamine receptor antagonist (domperidone: 1 mg/kg) resulted in the blockage of AMPA-mediated inhibition of PRL secretion, thus suggesting that this action is probably mediated by an increase in DA activity. In conclusion, we provide evidence for the physiological role of AMPA receptors in the control of PRL and GH secretion in prepubertal rats. In contrast, our data cast doubts on the involvement of AMPA receptors in the regulation of gonadotropin secretion at this age. The effects of AMPA reported herein were not mediated through activation of kainate receptors and were probably exerted at the hypothalamic or suprahypothalamic levels. In addition, we show that ovarian secretion actively modulates the effects of AMPA receptor activation on anterior pituitary secretion in prepubertal female rats.

Polychlorinated biphenyls (PCBs) affect the hypothalamic-pituitary-gonadal axis in many vertebrates, changing the hormonal regulation of reproduction. To identify one of the possible sites of action of PCBs on gonadotropin release in common carp, the direct effects of Aroclor 1254 on luteinizing hormone (LH) secretion from dispersed pituitary cells were investigated. Pituitary cells were obtained from sexually mature male and female common carp (Cyprinus carpio L.) at the time of natural spawning. The cells were incubated with different concentrations of Aroclor 1254 (5, 10, 50 and 100 ng mL–1 medium) and/or salmon gonadotropin-releasing hormone analogue (sGnRHa) at a concentration of 10−8 M. LH levels were measured in the cultured medium by the ELISA method after 10 hours of cell incubation. Incubation of male pituitary cells in the presence of tested concentrations of Aroclor did not change the basal LH secretion to the media. In the female pituitary cell incubations Aroclor (5, 10 and 100 ng mL–1 medium) caused a significant increase in LH concentrations in comparison to control incubations. In the case of sGnRHastimulated LH secretion in incubations of cells of both sexes, all the concentrations of Aroclor significantly stimulated LH release and potentiated stimulatory effects of sGnRH analogue. These results indicate that endocrine disrupters, such as Aroclor 1254, may affect reproduction in fish, acting also directly on gonadotrophs at the level of the pituitary gland, changing LH secretion.

The aim of the present study was to purify the common native carp growth hormone (ncGH), produce monoclonal antibodies (mAbs) to common native carp growth hormone (ncGH), and further enhance the sensitivity of enzyme-linked immunosorbent assays (ELISA) for ncGH. Additionally, we investigated changes in serum ncGH levels in carps raised in different environmental conditions. The recombinant grass carp (Ctenopharyngodon idella) growth hormone was purified and used as antigen to immunize the rabbit. The natural ncGH was isolated from the pituitaries of common carp. SDS-PAGE and Western blot utilizing the polyclonal anti-rgcGH antibody confirmed the purification of ncGH from pituitaries. Purified ncGH was then used as an immunogen in the B lymphocyte hybridoma technique. A total of 14 hybridoma cell lines (FMU-cGH 1-14) were established that were able to stably secrete mAbs against ncGH. Among them, eight clones (FMU-cGH1-6, 12 and 13) were successfully used for Western blot while nine clones (FMU-cGH 1-7, 9 and 10) were used in fluorescent staining and immunohistochemistry. Epitope mapping by competitive ELISA demonstrated that these mAbs recognized five different epitopes. A sensitive sandwich ELISA for detection of ncGH was developed using FMU-cGH12 as the coating mAb and FMU-cGH6 as the enzyme labeled mAb. This detection system was found to be highly stable and sensitive, with detection levels of 70 pg/mL. Additionally, we found that serum ncGH levels in restricted food group and in the net cage group increased 6.9-and 5.8-fold, respectively, when compared to controls, demonstrating differences in the GH stress response in common carp under different living conditions.

Estradiol Stimulates Growth Hormone Production in Female Goldfish

Dietary supplementation of olive leaf extract enhances growth performance, digestive enzyme activity and growth related genes expression in common carp Cyprinus carpio