Fertility and motility of sperm from Atlantic halibut (Hippoglossus hippoglossus) in relation to dose and timing of gonadotrophin-releasing hormone agonist implant

In broodstocks of Atlantic halibut, Hippoglossus hippoglossus, male and female gamete production often becomes unsynchronised towards the end of the spawning season—milt becomes very viscous and difficult to express while the females are still producing batches of good quality eggs. Gonadotrophin-releasing hormone agonist (GnRHa) has been shown to stimulate spermiation in a number of fish species. Therefore, we conducted two experiments where male halibut were implanted intramuscularly with pellets containing GnRHa. The effect of the pellets was tested at three periods: before, at the height of and at the end of spermiation. In the middle period, GnRHa was tested at two doses (5 and 25 μg/kg bodyweight). Measurements were made of milt hydration, sperm motility and fertilisation rate. Implanted males began spermiation at least 4 weeks before control males. Both doses of GnRHa increased the fluidity of the milt. This effect lasted for at least 20 days in the low dose group and for 40 days in the high dose group. When applied at the end of the season, GnRHa reversed the normal trend for the milt to become more viscous. GnRHa treatments did not affect fertilisation rates obtained with the sperm. However, towards the end of the spawning season, sperm motility was enhanced in males treated with the high dose of GnRHa (25 μg/kg) compared to controls. As described previously, plasma concentrations of the gonadal steroids, 5β-pregnane-3β,17,20β-triol 20-sulphate and 17,20-dihydroxy-4-pregnen-3-one, were significantly enhanced by GnRHa treatment. Concentrations of testosterone on the other hand decreased when spermiating males were treated with GnRHa. Our data suggest that 17,20β-dihydroxy-4-pregnen-3-one or its metabolites are involved in milt hydration, possibly through affecting ion transport.     

Ovulation and spawning induction in Caspian kutum Rutilus frisii kutum by administration of GnRHa and catecholaminergic pharmaceutical compounds with Ovaprim

The acceleration of final oocyte maturation and ovulation is recognized as a principal process in Caspian kutum Rutilus frisii kutum, a commercial and valuable species in Caspian Sea. The aim of this study was to evaluate the effects of catecholaminergic pharmaceutics which include Salbutamol (β₂‐adrenergic receptor agonist), Metoprolol (β₁‐adrenergic receptor antagonist), Clozapine and Olanzapine (the third generation of D₄ and D₂ dopamine receptor antagonists) which can be more efficient than the first and second generations, in combination with Buserelin ([D‐Ser(tBu)⁶,Pro⁹‐NEt]‐GnRHa) and Ovaprim (D‐Ala⁶,Pro⁹‐Net)‐sGnRH+Domperidone on ovulation and spawning induction in Caspian kutum. One hundred eight adult females were injected once with Buserelin Acetate (BUS) 5 μg kg^?1 BW, Ovaprim 0.5 mL kg^?1 BW, Salbutamol Sulphate (SLB) 4 mg kg^?1 BW, Olanzapine (OLZ) 5 mg kg^?1 BW, Clozapine (CZ) 12 mg kg^?1 BW and Metoprolol Tartrate (MTP) 5 mg kg^?1 BW, being divided into 12 groups: group 1, intact (negative control); group 2, Ovaprim (positive control); group 3, OLZ+SLB; group 4, OLZ+MTP; group 5, OLZ+BUS; group 6, CZ+SLB; group 7, CZ+MTP; group 8, CZ+BUS; group 9, OLZ+BUS+SLB; group 10, OLZ+BUS+MTP; group 11, CZ+BUS+SLB and group 12, CZ+BUS+MTP (N = 9). The results showed that the highest mean value in ovulation success, ovulation index, fertilization success, relative fecundity and the number of eggs belonged to Ovaprim treatment. On the other hand, spawning was successful in OLZ+BUS+SLB and CZ+BUS treatments (P < 0.05), whereas it was lower in CZ+MTP, CZ+BUS+MTP and OLZ+BUS than the other treatments. Therefore, it could be concluded that Clozapine and Olanzapine potentiated the effect of Buserelin treatment in ovulation and spawning induction, while Metoprolol blocked the stimulatory effects of GnRHa, Clozapine and Olanzapine. Salbutamol also can amplify stimulatory effect of all mentioned pharmaceutics.     

Induced ovulation of yellow catfish (Pelteobagrus fulvidraco) using a combination of a gonadotrop‐releasing hormone analogue and domperidone

The effects of an intraperitoneal hormone injection of gonadotropin‐releasing hormone agonist (D‐Ala⁶, Pro⁹‐NEt GnRHa) alone or in combination with a dopamine antagonist, domperidone (DOM), on ovulation induction in yellow catfish Pelteobagrus fulvidraco were tested. The hormone treatments were as follows: 6 mg kg⁻¹ body weight (BW) of carp pituitary extract as a positive control, GnRHa 10, 20, 40 and 80 μg kg⁻¹ BW and a combination of GnRHa and DOM as follows: 10 μg+5 mg, 20 μg+10 mg, 40 μg+20 mg and 80 μg+40 mg kg⁻¹ BW. Physiological saline (0.7% NaCl) was used as a negative control. Significant differences in the ovulation ratio, latency period and ovulation index (OI) were observed among treatments (P<0.05). The combination of GnRHa and DOM at doses of 40 μg+20 mg kg⁻¹ BW had higher values of the ovulation ratio and OI, and a shorter latency period compared with other treatments. The highest OI in GnRHa treatments was only 56.67%, suggesting a dopaminergic tone on gonadotropin secretion in this fish at the pre‐ovulatory stage. Therefore, ovulation can be successfully induced in yellow catfish with 40 μg kg⁻¹ GnRHa+20 mg kg⁻¹ DOM without affecting the egg quality.     

Dopaminergic influence on gonadotropin secretion in white sturgeon (Acipenser transmontanus)

The effects of dopamine on the secretion of two sturgeon gonadotropins (stGTH I and stGTH II) in sexually mature male white sturgeon (Acipenser transmontanus) were evaluated. In Experiment I, sturgeon were given intraperitoneal injections of physiological saline (PS), dopamine (100 mg kg⁻¹), the gonadotropin releasing hormone analog D-Ala⁶-des-Gly¹⁰-GnRH ethylamide (GnRHA) (10 μg/kg⁻¹), and a combination of GnRHa and dopamine. Fish receiving only GnRHa had significantly higher (p < 0.05) concentrations of plasma stGTH I and stGTH II compared to fish receiving PS, dopamine, or a combination of GnRHa and dopamine. Two hours following its administration, dopamine was effective in decreasing plasma concentrations of both stGTHs that were previously elevated by GnRHa. Dopamine or PS administered by themselves did not alter plasma concentrations of either stGTH. In Experiment II, sturgeon injected intraperitoneally with a combination of GnRHa and pimozide had significantly higher (p < 0.05) concentrations of plasma stGTH I and stGTH II compared to males receiving GnRHa or pimozide alone. While this effect of GnRHAa + pimozide was observed in the spring, no such potentiation was seen in these fish during the summer (Experiment III). These results represent the first evidence of dopaminergic inhibition of GnRH-induced pituitary gonadotropin secretion in Chondrostean fish.     

Effects of synthetic kisspeptin peptides and GnRH analogue on oocyte growth and circulating sex steroids in prepubertal female chub mackerel (Scomber japonicus)

In recent years, kisspeptin peptides, encoded by kiss genes have been used to manipulate reproductive processes in farmed animals, including fish. Our previous studies demonstrated that the chub mackerel brain expresses kiss1 and kiss2 and intramuscular injection of synthetic Kiss1 pentadecapeptide (Kiss1‐15) but not Kiss2 dodecapeptide (Kiss2‐12) accelerates spermatogenesis in prepubertal male chub mackerel (Scomber japonicus). In the present study, we evaluated their effects in prepubertal female chub mackerel. The gonadosomatic index (GSI) values of experimental fish did not show any significant changes. Condition factor (CF) values increased significantly in Kiss1‐15 treated fish, in comparison with control and GnRH analogue (GnRHa) injected fish. Histologically, only perinucleolar oocytes were found in all experimental fish. However, Kiss and GnRHa treated fish showed a significant increase in the perinucleolar oocyte diameter, in comparison with the control fish. Gene expression analyses revealed decreased expression of gnrh1 in the telencephalon‐preoptic region of the brain of Kiss2‐12 and GnRHa injected fish, in comparison with control fish. In contrast, GnRHa injected fish exhibited higher levels of fshβ in the pituitary, with no changes in the levels of lhβ among different treatments. Levels of circulating sex steroids, testosterone, and estradiol‐17β were significantly higher in Kiss1‐15 injected fish, in comparison with control fish. These results indicate that synthetic kisspeptin peptides and GnRHa can induce oocyte growth in prepubertal female chub mackerel.     

Gonadotrophin-releasing hormone agonist raises plasma concentrations of progestogens and enhances milt fluidity in male Atlantic halibut (Hippoglossus hippoglossus)

In two separate spawning seasons, spermiating male Atlantic halibut were implanted with pellets containing gonadotrophin-releasing hormone agonist (GnRHa). Males were bled repeatedly, and milt samples were collected. Blood samples were assayed for free and conjugated steroids: testosterone, 11-ketotestosterone, 17,20-dihydroxy-4-pregnen-3-one (17,20-P), 17,20-dihydroxy-4-pregnen-3-one (17,20-P), 17,20,21-trihydroxy-4-pregnen-3-one and steroids with a 17,20 configuration. Towards the end of the first season, pellets were implanted into three wild-caught and three hatchery-reared males. No control fish were available. The major progestogen in plasma was identified as sulphated 5-pregnane-3,17,20-triol (3,17,20-P-5-S). Concentrations of this steroid were stimulated by the GnRHa. Sulphated 17,20-P was also identified in the plasma, but at 10-fold lower concentrations than 3,17,20-P-5-S. In the middle of the second season, pellets were implanted into five hatchery-reared males; five unimplanted males were used as controls. Levels of androgens fell following GnRHa treatment, levels of progestogens rose briefly, and there was a significant increase in the fluidity of the milt. Of all the measured steroids, free and sulphated 17,20-P showed the best correlation with milt fluidity.     

Gonadotropin releasing hormone‐analogue (GnRHa) treatment improves the milt production and sperm motility of endangered Caspian brown trout,Salmo trutta caspius,over the course of a spawning season

To determine the effect of gonadotropin releasing hormone‐analogue (GnRHa) treatment on the milt quality of endangered Caspian brown trout, Salmo trutta caspius, the sperm motility (percentage and duration of motility), sperm production (sperm density, spermatocrit and milt volume) and milt pH were measured for GnRHa‐treated (the treatment group) and untreated groups (the control group) during the spawning season. For untreated brooders, the values of the motility per cent, sperm density and spermatocrit decreased continuously during the spawning season while the milt volume, duration of motility and milt pH showed only a significant decrease at the end of the season. For GnRHa‐treated males, these parameters increased 14 days after GnRHa treatment (first milt collection) and then decreased continuously towards the end of the season. In addition, the values of milt and sperm density yielded per treated male were higher than that in the untreated group, although these were not statistically different. In any case, the total sum of yielded milt from the treatment group over the spawning season was higher than that in the untreated group. In this experiment, significant positive correlations were found between milt parameters as follows: sperm motility vs. milt pH; sperm density vs. spermatocrit; milt volume vs. spermatocrit; and milt volume vs. sperm density. The results show that the treatment of Caspian brown trout by GnRHa can improve the milt quality in terms of sperm motility and sperm production during a spawning season.