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Development and validation of enzyme-linked immunosorbent assays specific for follicle-stimulating hormone and luteinizing hormone in Japanese eel

  • Yukinori KazetoEmail author
  • Toshiomi Tanaka
  • Hiroshi Suzuki
  • Yuichi Ozaki
  • Haruhisa Fukada
  • Koichiro Gen
Original Article Aquaculture

Abstract

Two types of gonadotropins (Gths), follicle-stimulating hormone (Fsh) and luteinizing hormone (Lh), are primary regulators of reproduction in vertebrates, including teleost fish. In this study, recombinant Japanese eel Gths (reGth; reFsh and reLh), and recombinant chimeric Gths (rcGth; rcFsh and rcLh), consisting of an eel β-subunit with rabbit glycoprotein hormone α-subunit, were produced as single-chain proteins, using a mammalian expression system and subsequently highly purified. Antibodies raised against rcGths were affinity purified with the corresponding reGth, and specific enzyme-linked immunosorbent assays (ELISAs) for Japanese eel Fsh and Lh were developed using these antibodies for the first time. The ELISAs were sensitive and parallelism was seen between the standard curve and serial dilutions of Japanese eel serum and pituitary extract (PE). Contents of Fsh and Lh protein in the pituitaries of the female Japanese eel at the pre-vitellogenic stage were comparable and the Lh contents at the migratory nucleus stage were highly induced during artificial induction of maturation by injections of salmon PE. Furthermore, the accumulated Lh was released at a high level by administration of a Lh-releasing hormone analogue and pimozide. Recombinant Gths and homologous ELISAs established for Japanese eel may be useful tools for studying reproductive eel biology.

Keywords

Gonadotropin Reproductive regulation Anguilla japonica Maturation 

Notes

Acknowledgements

We would like to express our appreciation to the staff of the National Research Institute of Aquaculture for the maintenance of the fish. This research was supported by the Ministry of Science and Technology (grant no. 22580219).

Funding

Funding was received from the Ministry of Science and Technology (grant no. 22580219).

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Copyright information

© Japanese Society of Fisheries Science 2019

Authors and Affiliations

  • Yukinori Kazeto
    • 1
    Email author
  • Toshiomi Tanaka
    • 2
  • Hiroshi Suzuki
    • 3
  • Yuichi Ozaki
    • 3
  • Haruhisa Fukada
    • 4
  • Koichiro Gen
    • 5
  1. 1.Kamiura Station, National Research Institute of AquacultureJapan Fisheries Research and Education AgencySaikiJapan
  2. 2.Hamanako BranchShizuoka Prefectural Research Institute of FisheryHamamatsuJapan
  3. 3.Tamaki Station, National Research Institute of AquacultureJapan Fisheries Research and Education AgencyWataraiJapan
  4. 4.Faculty of AgricultureKochi UniversityNankokuJapan
  5. 5.Seikai National Fisheries Research InstituteJapan Fisheries Research and Education AgencyNagasakiJapan

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