A dot-blot immunoassay for the detection of vitellogenin (Vtg) in plasma of adult grey mullet (Mugil cephalus) was developed. The assay identified the sex of the tested fish prior to detectable gonadal development, enabling the establishment of broodstock at the desired ratio of 7:4 females to males. This broodstock was maintained under natural photoperiod, and used to study the relative effect of gonadotropin-releasing hormone (GnRH) and dopamine antagonists on oocyte maturation and ovulation, as well as the effect of 17α-methyltestosterone (MT) on spermiation. Three groups of females were treated with: (i) a single injection of dopamine antagonist, domperidone (Dom), (ii) GnRH analog (GnRHa) administered via ethylene-vinyl acetate copolymer (EVAc) slow-release implants or (iii) a combination of both Dom and GnRHa. Males were treated with MT, administered via EVAc slow-release implants. An additional group of untreated fish was used as a control. The Dom treatment proved to be more potent than the GnRHa treatment, and did not differ significantly from the combined treatment. The Dom and Dom + GnRHa treatments accelerated oocyte development and increased plasma estradiol levels equally, whereas the GnRHa treatment did not vary significantly from the control. MT was found to be a potent spermiating agent, which enhanced steady milt production in all treated males. In contrast, no spontaneous spermiation occurred in untreated males. Plasma 11-ketotestosterone (11-KT) levels were significantly higher in MT-treated males than in the controls. Interestingly, MT-treated males held with the GnRHa + Dom-treated females showed higher levels of plasma 11-KT than those held with GnRHa-treated females, indicating an additive effect which is probably attributable to female pheromones. Fully mature females were induced to spawn by injecting GnRHa alone or coupled with metaclopramide (a dopamine D2 receptors antagonist). The combined treatment, which included a dopamine antagonist, was found to be more potent in inducing ovulation and spawning as compared to GnRHa alone. In conclusion, our data suggest that dopaminergic inhibition is a major barrier along the reproductive axis that arrests spontaneous spawning in captive mullets.
Bibliographical noteFunding Information:
The authors are indebted to Dr. E. Lubzens (IOLR, Haifa, Israel) for providing the antisera raised against seabream vitellogenin, and to Dr. David Kime (Sheffield, UK) for providing the detailed ELISA protocol, as well as the 11-KT antiserum. Special thanks to Dr. A. Elizur and Dr. A. Colorni for reviewing the manuscript. This work was supported by a Grant 894-0143-03 from the Israeli Ministry of Agriculture and by a Grant TA-MOU-99-M18-061 from MERC-US AID.
- Gonadotropin-releasing hormone
- Mugil cephalus
- Spawning induction