Hostname: page-component-586b7cd67f-l7hp2 Total loading time: 0 Render date: 2024-11-22T21:02:23.503Z Has data issue: false hasContentIssue false
  • English
  • Français

Successful vitrification of whole juvenile testis in the critically endangered cyprinid honmoroko (Gnathopogon caerulescens)

Published online by Cambridge University Press:  24 August 2017

Shogo Higaki ,
Natsue Kuwata ,
Kotaro Tanaka ,
Ikuo Tooyama ,
Yasuhiro Fujioka ,
Noriyoshi Sakai  and
Tatsuyuki Takada
Shogo Higaki
Affiliation:
Laboratory of Cell Engineering, Department of Pharmaceutical Sciences, Ritsumeikan University, Kusatsu, Shiga 525–8577, Japan.
Natsue Kuwata
Affiliation:
Laboratory of Cell Engineering, Department of Pharmaceutical Sciences, Ritsumeikan University, Kusatsu, Shiga 525–8577, Japan.
Kotaro Tanaka
Affiliation:
Laboratory of Cell Engineering, Department of Pharmaceutical Sciences, Ritsumeikan University, Kusatsu, Shiga 525–8577, Japan.
Ikuo Tooyama
Affiliation:
Molecular Neuroscience Research Center, Shiga University of Medical Science, Otsu, Shiga, 520–2192, Japan.
Yasuhiro Fujioka
Affiliation:
Lake Biwa Museum, Oroshimo 1091, Kusatsu, Shiga 525-0001, Japan.
Noriyoshi Sakai
Affiliation:
Genetic Strains Research Center, National Institute of Genetics, Mishima, Shizuoka 411–8540, Japan.
Tatsuyuki Takada*
Affiliation:
Laboratory of Cell Engineering, Department of Pharmaceutical Sciences, Ritsumeikan University, Nojihigashi 1-1-1, Kusatsu, Shiga 525–8577, Japan.
*
All correspondence to: Tatsuyuki Takada. Laboratory of Cell Engineering, Department of Pharmaceutical Sciences, Ritsumeikan University, Nojihigashi 1-1-1, Kusatsu, Shiga 525–8577, Japan. Tel: +81 77 561 2569. Fax: +81 77 561 2569. E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Summary

Sperm cryopreservation is a valuable conservation method for endangered fish species. Here we report an easy and efficient cryopreservation method for juvenile whole testis by vitrification and successful sperm production from the vitrified whole testis via in vitro spermatogenesis in the critically endangered cyprinid honmoroko (Gnathopogon caerulescens). Juvenile testis (approximately 10 mm in length and 1 mm in width), consisting predominantly of spermatogonia, were aseptically dissected out and adherent fatty and non-testicular tissues were subsequently removed. Then, the testes were rapidly cooled on a nylon mesh by direct immersion in liquid nitrogen after serial exposures to pretreatment solution (PS), containing 2 M ethylene glycol (EG) and 1 M dimethyl sulfoxide (DMSO), for 20 or 30 min and vitrification solution (VS), containing 3 M EG, 2 M DMSO, and 0.5 M sucrose, for 5, 10, or 20 min. The highest survival rate of testicular cells (84.0%) was obtained from testes vitrified by immersion in PS for 20 min and in VS for 10 min. Spermatogonia were recovered from the vitrified testis by dissociation and cell culture produced many haploid sperm. Fertility and developmental competence were confirmed by in vitro fertilization assays. These results indicate that the vitrification of juvenile whole testis provides a new strategy to preserve the genetic resources of endangered fishes without affecting their reproductive population.

Keywords

CryopreservationEndangered fishIn vitro spermatogenesisSpermatogoniaWhole testis vitrification
Type
Research Article
Information
Zygote , Volume 25 , Issue 5 , October 2017 , pp. 652 - 661
Copyright
Copyright © Cambridge University Press 2017 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Amorim, C.A., Curaba, M., Van Langendonckt, A., Dolmans, M.-M. & Donnez, J. (2011). Vitrification as an alternative means of cryopreserving ovarian tissue. Reprod. Biomed. Online 23, 160–86.Google Scholar
Brockbank, K.G. & Taylor, M.J. (2006). Tissue preservation. In Advances in Biopreservation (ed. Baust, J.), pp. 157–96. London: CRC Press.Google Scholar
Fowler, A. & Toner, M. (2005). Cryo-injury and biopreservation. Ann. N. Y. Acad. Sci. 1066, 119–35.Google Scholar
Fujioka, Y. (1993). Induction of gynogenetic diploids and cytological studies in honmoroko Gnathopogon caerulescens. Nippon Suisan Gakkaishi 59, 493–500.Google Scholar
Fujioka, T., Yasuchika, K., Nakamura, Y., Nakatsuji, N. & Suemori, H. (2004). A simple and efficient cryopreservation method for primate embryonic stem cells. Int. J. Dev. Biol. 48, 1149–54.Google Scholar
Higaki, S., Kawakami, Y., Eto, Y., Yamaha, E., Nagano, M., Katagiri, S., Takada, T. & Takahashi, Y. (2013). Cryopreservation of zebrafish (Danio rerio) primordial germ cells by vitrification of yolk-intact and yolk-depleted embryos using various cryoprotectant solutions. Cryobiology 67, 374–82.Google Scholar
Higaki, S., Shimada, M., Kawamoto, K., Todo, T., Kawasaki, T., Tooyama, I., Fujioka, Y., Sakai, N. & Takada, T. (2017). In. vitro differentiation of fertile sperm from cryopreserved spermatogonia in endangered endemic cyprinid honmoroko (Gnathopogon caerulescens). Sci. Rep. 7, 42852.Google Scholar
Leal, M.C., Cardoso, E.R., Nóbrega, R.H., Batlouni, S.R., Bogerd, J., França, L.R. & Schulz, R.W. (2009). Histological and stereological evaluation of zebrafish (Danio rerio) spermatogenesis with an emphasis on spermatogonial generations. Biol. Reprod. 81, 177–87.Google Scholar
Lee, S., Iwasaki, Y., Shikina, S. & Yoshizaki, G. (2013). Generation of functional eggs and sperm from cryopreserved whole testes. Proc. Natl. Acad. Sci. USA 110, 1640–5.Google Scholar
Luyet, B. & Hodapp, E. (1938). Revival of frog's spermatozoa vitrified in liquid air. Exp. Biol. Med. 39, 433–4.Google Scholar
Maki, I. (1968). Studies on the population dynamics of Gnathopogon. caerulescens sauvage (pisces), in Lake Biwa, Japan. Jap. J. Ecol. 18, 158–66 [in Japanese].Google Scholar
Mazur, P. (1984). Freezing of living cells: mechanisms and implications. Am. J. Physiol. Cell. Physiol. 247, 125–42.Google Scholar
Ministry of the Environment Government of Japan, Red List of Japanese Brackish and Freshwater Fishes 2015, (accessed 28 May 2017) http://www.env.go.jp/press/files/jp/28060.pdf Google Scholar
Nakamura, M. (1969). Honomoroko. In Cyprinid Fishes of Japan: Studies on the Life History of Cyprinid Fishes of Japan (ed. Nakamura, M.) pp. 117–25. Tokyo: Research Institute for Natural Resources, [in Japanese].Google Scholar
Okutsu, T., Shikina, S., Kanno, M., Takeuchi, Y. & Yoshizaki, G. (2007). Production of trout offspring from triploid salmon parents. Science 317, 1517.Google Scholar
Pšenička, M., Saito, T., Rodina, M. & Dzyuba, B. (2016). Cryopreservation of early stage Siberian sturgeon Acipenser baerii germ cells, comparison of whole tissue and dissociated cells. Cryobiology 72, 119–22.Google Scholar
Raz, E. (2000). The function and regulation of vasa-like genes in germ-cell development. Genome Biol. 1, 1017.Google Scholar
Sakai, N. (2002). Transmeiotic differentiation of zebrafish germ cells into functional sperm in culture. Development 129, 3359–65.Google Scholar
Sakai, N (2006). In vitro male germ cell cultures of zebrafish. Methods 39, 239–45.Google Scholar
Saragusty, J. & Arav, A. (2011). Current progress in oocyte and embryo cryopreservation by slow freezing and vitrification. Reproduction 141, 1–19.Google Scholar
Scheffen, B. (1986). A simple and efficient procedure for preservation of mouse embryos by vitrification. Cryo-letters 7, 260–9.Google Scholar
Seki, S., Kusano, K., Lee, S., Iwasaki, Y., Yagisawa, M., Ishida, M., Hiratsuka, T., Sasado, T., Naruse, K. & Yoshizaki, G. (2017). Production of the medaka derived from vitrified whole testes by germ cell transplantation. Sci. Rep. 3, 43185.CrossRefGoogle Scholar
Tiersch, T. (2001). Cryopreservation in aquarium fishes. Mar. Biotechnol. 3, S212–23.Google Scholar
Westerfield, M. (2007). Recipes-embryo extract. In The Zebrafish Book (ed. Westerfield, M.), p. 10.14. Eugene, Oregon, USA: The University of Oregon Press.Google Scholar
Yang, H. & Tiersch, T. (2009). Current status of sperm cryopreservation in biomedical research fish models: Zebrafish, medaka & Xiphophorus . Comp. Biochem. Physiol. C. Pharmacol. Toxicol. Endocrinol. 149, 224–32.Google Scholar
Yavin, S. & Arav, A. (2007). Measurement of essential physical properties of vitrification solutions. Theriogenology 67, 81–9.Google Scholar
Supplementary material: Image

Higaki et al. supplementary material

Higaki et al. supplementary material 1

Download Higaki et al. supplementary material(Image)
Image 3 MB

Higaki et al. supplementary material

Higaki et al. supplementary material 2

Download Higaki et al. supplementary material(Video)
Video 738.1 KB

Higaki et al. supplementary material

Higaki et al. supplementary material 3

Download Higaki et al. supplementary material(Video)
Video 462.5 KB
Supplementary material: File

Higaki et al. supplementary material

Higaki et al. supplementary material 4

Download Higaki et al. supplementary material(File)
File 246.8 KB