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Intra-ooplasmic injection of a multiple number of sperm to induce androgenesis and polyploidy in the dojo loach Misgurnus anguillicaudatus (Teleostei: Cobitidae)

Published online by Cambridge University Press:  29 October 2018

George Shigueki Yasui ,
Taiju Saito ,
Yan Zhao ,
Takafumi Fujimoto Open the ORCID record for Takafumi Fujimoto [Opens in a new window] ,
Etsuro Yamaha  and
Katsutoshi Arai
George Shigueki Yasui
Affiliation:
Faculty and Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido, Japan;
Taiju Saito
Affiliation:
Nanae Fresh-Water Laboratory, Field Science Center for Northern Biosphere, Sakura, Nanae, Kameda, Hokkaido, Japan
Yan Zhao
Affiliation:
Faculty and Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido, Japan;
Takafumi Fujimoto*
Affiliation:
Faculty and Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido, Japan;
Etsuro Yamaha
Affiliation:
Nanae Fresh-Water Laboratory, Field Science Center for Northern Biosphere, Sakura, Nanae, Kameda, Hokkaido, Japan
Katsutoshi Arai
Affiliation:
Faculty and Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido, Japan;
*
*Address for correspondence: Takafumi Fujimoto. Faculty and Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido, Japan. Tel:/Fax: +81 138 40 5536. E-mail: [email protected].
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Summary

Polyspermy was initiated by microinjecting a multiple number of sperm into the activated and dechorionated eggs of dojo loach Misgurnus anguillicaudatus (Teleostei: Cobitidae). A 10-nl sperm suspension from an albino (recessive trait) male (105, 106, 107 or 108 sperm ml −1) was microinjected into eggs from a wild-type female. Although the rates of embryos developing into the blastula stage in the injection group at the highest sperm concentration were similar to that of the control group, the hatching rates of the injection group were much lower. A large proportion of embryos that developed from the injected eggs was haploid and were mosaics containing haploid cells. Most of the haploid and mosaic embryos inherited only paternally derived alleles in the microsatellite markers (i.e. androgenesis was initiated by injecting multiple sperm). In contrast, some haploid embryos contained both paternal and maternal alleles despite haploidy, suggesting that they were mosaics consisting of cells with either paternal or maternal inheritance. The injected eggs displayed diploid, hypotriploid and triploid cells, all of which included both maternally and paternally derived alleles. One albino tetraploid with only paternal alleles was also observed from the injected eggs. These results suggested that part of the sperm microinjected into the ooplasm should form a male pronucleus/pronuclei, which could develop by androgenesis or could fuse with the female pronucleus/pronuclei. Therefore, microinjection of multiple sperm should be considered a potential technique to induce androgenesis and polyploidy.

Keywords

FertilizationFishICSIPronucleusSpermatozoa
Type
Research Article
Information
Zygote , Volume 26 , Issue 5 , October 2018 , pp. 408 - 416
Copyright
© Cambridge University Press 2018 

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Footnotes

*

Present address: Laboratory of Fish Biotechnology, National Center for Research and Conservation of Continental Fish, Chico Mendes Institute of Biodiversity Conservation, Rodovia Pref. Euberto Nemésio Pereira de Godoy, Pirassununga, Säo Paulo State, Brazil.

†

Present address: South Ehime Fisheries Research Center, Ehime University, Uchidomari, Ainan, Ehime, Japan.

‡

Present address: College of Fisheries and Life Science, Shanghai Ocean University, Shanghai, China.

References

Fujimoto, T, Kataoka, T, Sakao, S, Saito, T, Yamaha, E and Arai, K (2006) Developmental stages and germ cell lineage of the loach (Misgurnus anguillicaudatus). Zool Sci 23, 977–989.Google Scholar
Fujimoto, T, Sakao, S, Yamaha, E and Arai, K (2007) Evaluation of different doses of UV irradiation to loach eggs for genetic inactivation of the maternal genome. J Exp Zool Part A 307, 449–462.Google Scholar
Kurokura, H, Hirano, R, Tomita, M and Iwahashi, M (1984) Cryopreservation of carp sperm. Aquaculture 37, 267–273.Google Scholar
Mazur, P, Leibo, SP and Seidel, GE Jr (2008) Cryopreservation of the germplasm of animals used in biological and medical research: importance, impact, status, and future directions. Biol Reprod 78, 2–12.Google Scholar
Morishima, K, Nakayama, I and Arai, K (2008) Genetic linkage map of the loach Misgurnus anguillicaudatus (Teleostei: Cobitidae). Genetica 132, 227–241.Google Scholar
Morozumi, K, Shikano, T, Miyazaki, S and Yanagimachi, R (2006) Simultaneous removal of sperm plasma membrane and acrosome before intracytoplasmic sperm injection improves oocyte activation/embryonic development. Proc Natl Acad Sci USA 103, 17661–17666.Google Scholar
Otani, S, Iwai, T, Nakahata, S, Sakai, C and Yamashita, M (2009) Artificial fertilization by intracytoplasmic sperm injection in a teleost fish, the medaka (Oryzias latipes). Biol Reprod 80, 175–183.Google Scholar
Poleo, GA, Denniston, RS, Reggio, BC, Godke, RA and Tiersch, TR (2001) Fertilization of eggs of zebrafish, Danio rerio, by intracytoplasmic sperm injection. Biol Reprod 65, 961–966.Google Scholar
Poleo, GA, Godke, RA and Tiersch, TR (2005) Intracytoplasmic sperm injection using cryopreserved, fixed, and freeze-dried sperm in eggs of Nile tilapia. Mar Biotechnol 7, 104–111.Google Scholar
Saito, T, Fujimoto, T, Maegawa, S, Inoue, K, Arai, K and Yamaha, E (2006) Visualization of primordial germ cells in vivo using GFP-nos1 3′UTR mRNA. Int J Dev Biol 50, 691–699.Google Scholar
Tanaka, D, Takahashi, A and Ueno, K (2009) Morphometric characteristics and reproductive capacity of nuclear transplants derived from embryonic cells of loach, Misgurnus anguillicaudatus, J Exp Zool 311A, 11–19.Google Scholar
Wakayama, T and Yanagimachi, R (1998) Development of normal mice from oocytes injected with freeze-dried spermatozoa. Nat Biotechnol 16, 639–641.Google Scholar
Yamaha, E, Usui, K, Onozato, H and Hamada, K (1986) A method for dechorionation in goldfish Carassius auratus . Bull Japan Soc Sci Fish 52, 1929–1934.Google Scholar
Yamashita, M, Onozato, H, Nakanishi, T and Nagahama, Y (1990) Breakdown of the sperm nuclear envelope is a prerequisite for male pronucleus formation: direct evidence from the gynogenetic crucian carp Carassius auratus langsdorfii . Dev Biol 137, 155–160.Google Scholar
Yanagimachi, R (2005) Intracytoplasmic injection of spermatozoa and spermatogenic cells: Its biology and applications in humans and animals. Reprod Biomed Online 10, 247–288.Google Scholar
Yasui, GS, Arias-Rodriguez, L, Fujimoto, T and Arai, K (2009) A sperm cryopreservation protocol the loach Misgurnus anguillicaudatus and its applicability for other related species. Anim Reprod Sci 116, 335–345.Google Scholar