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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: fujimoto@fish.hokudai.ac.jp.
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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.

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