Regulation of Gonadotropin-Releasing Hormone (GnRH)-Receptor Gene Expression in Tilapia: Effect of GnRH and Dopamine

The present work was designed to study certain aspects of the endocrine regulation of gonadotropin-releasing hormone receptor (GnRH-R) in the pituitary of the teleost fish tilapia. A GnRH-R was cloned from the pituitary of hybrid tilapia (taGnRH-R) and was identified as a typical seven-transmembrane receptor. Northern blot analysis revealed a single GnRH-R transcript in the pituitary of approximately 2.3 kilobases. The taGnRH-R mRNA levels were significantly higher in females than in males. Injection of the salmon GnRH analog (sGnRHa; 5–50 μg/kg) increased the steady-state levels of taGnRH-R mRNA, with the highest response recorded at 25 μg/kg and at 36 h. At the higher dose of sGnRHa (50 μg/kg), taGnRH-R transcript appeared to be down-regulated. Exposure of tilapia pituitary cells in culture to graded doses (0.1–100 nM) of seabream (sbGnRH = GnRH I), chicken II (cGnRH II), or salmon GnRH (sGnRH = GnRH III) resulted in a significant increase in taGnRH-R mRNA levels. The highest levels of both LH release and taGnRH-R mRNA levels were recorded after exposure to cGnRH II and the lowest after exposure to sbGnRH. The dopamine-agonist quinpirole suppressed LH release and mRNA levels of taGnRH-R, indicating an inhibitory effect on GnRH-R synthesis. Collectively, these data provide evidence that GnRH in tilapia can up- regulate, whereas dopamine down-regulates, taGnRH-R mRNA levels

Effects of a saponin fraction extracted from Trigonella foenum-graecum L. and two commercially available saponins on sex ratio and gonad histology of Nile tilapa fry, Oreochromis niloticus (L.)

Over three million tonnes (t) of tilapia, mostly Nile tilapia (Oreochromis niloticus, L.), are produced annually making it the second most abundantly produced freshwater fish (FAO, 2010). Tilapia are mouthbreeders that often produce stunted populations under pond conditions; one means of prevention is to produce all-male fish with the additional advantage that males usually grow faster than females. All-male populations can be achieved by supplementing feed with androgens such as 17-α-Methyltestosterone (MT) during days 10–25 post-hatch (Pandian and Sheela, 1995). However, MT is considered to be carcinogenic (Velazquez and Alter, 2004), and Hulak et al. (2008) also showed that effluents of systems in which carp were fed diets containing MT caused masculinization of female fish. Furthermore, in aquaculture the application of hormones to fish destined for human consumption is prohibited in the European Union under directive 96/22/EC, article 5, which also prohibits import of animal products produced with hormones. Kwon et al. (2000) showed that Fadrozole, a non-steroidal compound, caused masculinization in tilapia by inhibiting aromatase, which is the enzyme responsible for the conversion of endogenous androgens to estrogens. Steinbronn et al. (2004) were able to show that a dose of 2000 ppm Quillaja saponins (Sigma S-2149) inhibited reproduction of tilapia after dietary application for 32 days to first-feeding fry, suggesting saponins as a possible alternative to MT. These secondary plant compounds consist of either a steroid or triterpenoid basic structure (aglycone or sapogenin) plus one or more sugar side chains (Francis et al., 2002a). In a previous experiment a saponin fraction from the soapbark tree (Quillaja saponaria M.) inhibited aromatase in vitro (Golan et al., 2008). The fenugreek plant (Trigonella foenum-graecum L), widely cultivated in the Middle East and Asia, also has a high saponin content. The experiment was therefore conducted to test whether saponin fractions from Q. saponaria and from T. foenum-graecum were able to influence the sex ratio and gonad histology of Nile tilapia

Twin pregnancy with complete hydatidiform mole and coexisting fetus following ovulation induction with a non-prescribed clomiphene citrate regimen: a case report

<p>Abstract</p> <p>Introduction</p> <p>Twin pregnancy with complete hydatidiform mole represents a very rare obstetric problem. Management of such cases is always problematic because the possibility of fetal survival should always be weighed against the risk of complications of molar pregnancy.</p> <p>Case presentation</p> <p>A 34-year-old Caucasian woman presented to our center with mild vaginal bleeding. Our patient was 16 weeks pregnant after a seven-year period of primary infertility. She became pregnant following a non-prescribed regimen of clomiphene citrate extending from the second day to the 13th day of her last cycle. A transabdominal ultrasound examination revealed a twin pregnancy with complete hydatidiform mole and a coexisting fetus. Serum β human chorionic gonadotropin was falsely low as identified by serial dilution of the sample (the 'hook effect'). Our patient refused termination of pregnancy and she was hospitalized for strict observation and follow-up. Unfortunately, she developed an attack of severe vaginal bleeding and a hysterotomy was performed. The fetus died shortly after birth.</p> <p>Conclusions</p> <p>Twin pregnancy with complete hydatidiform mole represents a matter of controversy. We suggest that conservation should always be considered whenever tertiary care services and strict observation are available.</p

Effects of saponin fractions from fenugreek and the soap bark tree in the diet on performance of Nile tilapia, Oreochromis niloticus (L.)

Saponins are generally regarded as anti-nutritional factors in aquaculture diets. However, previous experiments have shown that low dietary levels of saponins derived from Quillaja saponaria Molina do have growth promoting effects on common carp and Nile tilapia. Based on these experiments, we conducted an experiment in which we fed eluated saponin fractions from Q. saponaria and Trigonella foenum-graecum L. (fenugreek) to Nile tilapia in a respirometric system allowing for continuous measurement of oxygen consumption. Saponins were eluated with consecutive methanol/water concentrations (v/v, 40/60, 60/40, 80/20) resulting in three different eluates for each plant. Fractions chosen were the 80% methanol eluate from Q. saponaria (80QS) and all three eluates from T. foenum-graecum (40TS, 60TS and 80TS). Three fish each were fed with low levels (150 mg kg-1 diet) of saponins in the diet and a control diet without saponins. Growth, feed and nutrient utilization, proximate composition, oxygen consumption and metabolic performance were evaluated. The fish grew between 224% (40TS) and 266% (Control) over the eight week period. Feed conversion ratios were between 0.94 (80TS) and 1.15 (40TS) and protein efficiency ratios between 2.54 (80TS) and 2.16 (40TS). Due to low sample sizes, no statistical differences were found between control fish and saponin fed fish. However, numerically one of the tested saponin fractions (40TS) showed inferior performance (Table 1). It is concluded that the tested saponins in the tested concentrations are no potential growth promoter for Nile tilapia. On the contrary, one fraction appears to be a growth inhibitor

Yolk utilization and growth during the early larval life of the Silver Perch, Bidyanus bidyanus (Mitchell, 1838)

The aim of this research was to investigate the yolk sac and oil globule utilization by silver perch (Bidyanus bidyanus) larvae produced from domesticated broodfish. The larvae were kept unfed in the holding tank, sampled, and investigated by image analysis software to determine various characteristics, such as the diameters of ova, water-hardened eggs, yolk-sac, oil globules, and the total length of larvae. The research illustrated that, with the exception of oil globule diameter, all other morphometric parameters were significantly lower (P &lt; 0.05) when compared to the larvae from the wild broodfish. The yolk sac was completely absorbed at 96 h post-hatching (hph) and the oil globule was visible until 240 hph. The larvae exhibited predatory movements and tried to catch rotifer at 4 days post hatching (dph). However, the onset of feeding took place at 5 dph, while 100% of feeding occurred at 6 dph. During the first 96 h (h), larvae grew significantly faster than the next 144 h. Larvae encountered low mortalities (&lt;10%) during the first 96 hph, before increasing significantly in the next 24 h and no unfed larvae survived post 240 h. The results also suggested that the exogenous feed should be available at 96 hph, which is well after the yolk sac is completely depleted. In addition, although most of eggs and larval performance from domesticated broodfish were inferior compared to the wild one, it has larger oil globule that could make longer of its mixed feeding period and therefore could have better in viability

Defining Global Gene Expression Changes of the Hypothalamic-Pituitary-Gonadal Axis in Female sGnRH-Antisense Transgenic Common Carp (Cyprinus carpio)

BACKGROUND: The hypothalamic-pituitary-gonadal (HPG) axis is critical in the development and regulation of reproduction in fish. The inhibition of neuropeptide gonadotropin-releasing hormone (GnRH) expression may diminish or severely hamper gonadal development due to it being the key regulator of the axis, and then provide a model for the comprehensive study of the expression patterns of genes with respect to the fish reproductive system. METHODOLOGY/PRINCIPAL FINDINGS: In a previous study we injected 342 fertilized eggs from the common carp (Cyprinus carpio) with a gene construct that expressed antisense sGnRH. Four years later, we found a total of 38 transgenic fish with abnormal or missing gonads. From this group we selected the 12 sterile females with abnormal ovaries in which we combined suppression subtractive hybridization (SSH) and cDNA microarray analysis to define changes in gene expression of the HPG axis in the present study. As a result, nine, 28, and 212 genes were separately identified as being differentially expressed in hypothalamus, pituitary, and ovary, of which 87 genes were novel. The number of down- and up-regulated genes was five and four (hypothalamus), 16 and 12 (pituitary), 119 and 93 (ovary), respectively. Functional analyses showed that these genes involved in several biological processes, such as biosynthesis, organogenesis, metabolism pathways, immune systems, transport links, and apoptosis. Within these categories, significant genes for neuropeptides, gonadotropins, metabolic, oogenesis and inflammatory factors were identified. CONCLUSIONS/SIGNIFICANCE: This study indicated the progressive scaling-up effect of hypothalamic sGnRH antisense on the pituitary and ovary receptors of female carp and provided comprehensive data with respect to global changes in gene expression throughout the HPG signaling pathway, contributing towards improving our understanding of the molecular mechanisms and regulative pathways in the reproductive system of teleost fish

Rapid Dopaminergic Modulation of the Fish Hypothalamic Transcriptome and Proteome

Background - Dopamine (DA) is a major neurotransmitter playing an important role in the regulation of vertebrate reproduction. We developed a novel method for the comparison of transcriptomic and proteomic data obtained from in vivo experiments designed to study the neuroendocrine actions of DA. // Methods and Findings - Female goldfish were injected (i.p.) with DA agonists (D1-specific; SKF 38393, or D2-specific; LY 171555) and sacrificed after 5 h. Serum LH levels were reduced by 57% and 75% by SKF 38393 and LY 171555, respectively, indicating that the treatments produced physiologically relevant responses in vivo. Bioinformatic strategies and a ray-finned fish database were established for microarray and iTRAQ proteomic analysis of the hypothalamus, revealing a total of 3088 mRNAs and 42 proteins as being differentially regulated by the treatments. Twenty one proteins and mRNAs corresponding to these proteins appeared on both lists. Many of the mRNAs and proteins affected by the treatments were grouped into the Gene Ontology categorizations of protein complex, signal transduction, response to stimulus, and regulation of cellular processes. There was a 57% and 14% directional agreement between the differentially-regulated mRNAs and proteins for SKF 38393 and LY 171555, respectively. // Conclusions - The results demonstrate the applicability of advanced high-throughput genomic and proteomic analyses in an amendable well-studied teleost model species whose genome has yet to be sequenced. We demonstrate that DA rapidly regulates multiple hypothalamic pathways and processes that are also known to be involved in pathologies of the central nervous system

Regulation of fish gonadotropins

10.1016/S0074-7696(05)25004-0International Review of Cytology225131-185IRCY