Abstract
Sturgeons are one of the oldest, biggest, most valuable and today also most endangered group of fish species. Germ stem cells (GSCs), such us embryonic primordial germ cells (PGCs) or spermatogonial/oogonial stem cells, can be a key for an effective conservation and possible restoration of these unique and astonishing fishes. In this chapter, labeling, development, isolation, and transplantation of GSCs were studied in sturgeons. It was shown that the maternally supplied germ plasm, which determines the PGC origin, is localized in vegetal pole of ovulated egg and remains there throughout the cleavage period, and therefore, the PGC specification pattern is similar to that of anuran amphibians rather than teleostean fishes. This knowledge enabled to develop an original PGC labeling method using common cell tracer dye injection into the vegetal pole of two- to eight-cell stage embryo. Next inhibition of maternally supplied dead end RNA resulted in PGC mismigration and general sterilization of individuals. This method enables preparation of recipients for germ cell transplantation. Isolation and transplantation of spermatogonia and oogonia were developed as well. It was tested that one sturgeon juvenile (Siberian sturgeon) can provide approximately one million germ cells suitable for transplantation. Moreover, it was shown that these cells are capable of propagation via an in vitro culture system and of cryopreservation. After freezing/thawing of sturgeon gonadal tissue followed by enzymatic dissociation, above 90% of viable cells were obtained and used for transplantation. The technique of surrogate production can be applied for conservation and possibly restoration of critically endangered sturgeon species with a long term of maturation and a big body size (e.g., beluga), whereas a more common species with shorter term of maturation and smaller body size (e.g., sterlet) can be used as a recipient (surrogate parent).
The chapter is written based on Pšenička’s inaugural dissertation.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Bellaiche J, Lareyre J-J, Cauty C, Yano A, Allemand I, Le Gac F (2014) Spermatogonial stem cell quest: nanos2, marker of a subpopulation of undifferentiated a spermatogonia in trout testis. Biol Reprod 90:79. https://doi.org/10.1095/biolreprod.113.116392
Bellve AR, Cavicchia JC, Millette CF, O’Brien DA, Bhatnagar YM, Dym M (1977) Spermatogenic cells of the prepubertal mouse. Isolation and morphological characterization. J Cell Biol 74:68–85. https://doi.org/10.1083/jcb.74.1.68
Blaser H, Eisenbeiss S, Neumann M, Reichman-Fried M, Thisse B, Thisse C, Raz E (2005) Transition from non-motile behaviour to directed migration during early PGC development in zebrafish. J Cell Sci 118:4027–4038. https://doi.org/10.1242/jcs.02522
Bontems F, Stein A, Marlow F, Lyautey J, Gupta T, Mullins MC, Dosch R (2009) Bucky ball organizes germ plasm assembly in zebrafish. Curr Biol 19:414–422. https://doi.org/10.1016/j.cub.2009.01.038
Brinster RL, Zimmermann JW (1994) Spermatogenesis following male germ-cell transplantation. Proc Natl Acad Sci U S A 91:11298–11302. https://doi.org/10.1073/pnas.91.24.11298
Chebanov MS, Galich EV (2011) Sturgeon hatchery manual, FAO fisheries and aquaculture technical paper 558. Food and Agriculture Organisation of the United Nations, Ankara
Doitsidou M, Reichman-Fried M, Stebler J, Koprunner M, Dorries J, Meyer D, Esguerra CV, Leung T, Raz E (2002) Guidance of primordial germ cell migration by the chemokine SDF-1. Cell 111:647–659. https://doi.org/10.1016/S0092-8674(02)01135-2
Eddy EM (1976) Germ plasm and the differentiation of the germ cell line. Int Rev Cytol 43:229–280. https://doi.org/10.1016/S0074-7696(08)60070-4
Ewen-Campen B, Schwager EE, Extavour CGM (2010) The molecular machinery of germ line specification. Mol Reprod Dev 77:3–18. https://doi.org/10.1002/mrd.21091
Extavour CG, Akam M (2003) Mechanisms of germ cell specification across the metazoans: epigenesis and preformation. Development 130:5869–5884. https://doi.org/10.1242/dev.00804
Fan L, Moon J, Wong T-T, Crodian J, Collodi P (2008) Zebrafish primordial germ cell cultures derived from vasa: RFP transgenic embryos. Stem Cells Dev 17:585–597. https://doi.org/10.1089/scd.2007.0178
Fatira E, Havelka M, Labbé C, Depincé A, Iegorova V, Pšenička M, Saito T (2018) Application of interspecific somatic cell nuclear transfer (iSCNT) in sturgeons and an unexpectedly produced gynogenetic sterlet with homozygous quadruple haploid. Sci Rep 8(1):5997
Flynn SR, Matsuoka M, Reith M, Martin-Robichaud DJ, Benfey TJ (2006) Gynogenesis and sex determination in shortnose sturgeon, Acipenser brevirostrum Lesuere. Aquaculture 253:721–727. https://doi.org/10.1016/j.aquaculture.2005.09.016
Havelka M, Hulák M, Ráb P, Rábová M, Lieckfeldt D, Ludwig A, Rodina M, Gela D, Pšenička M, Bytyutskyy D, Flajšhans M (2014) Fertility of a spontaneous hexaploid male Siberian sturgeon, Acipenser baerii. BMC Genet 15:5. https://doi.org/10.1186/1471-2156-15-5
Ihssen PE, McKay LR, McMillan I, Phillips RB (1990) Ploidy manipulation and gynogenesis in fishes: cytogenetic and fisheries applications. Trans Am Fish Soc 119:698–717. https://doi.org/10.1577/1548-8659(1990)119<0698:PMAGIF>2.3.CO;2
Ikenishi K, Kotani M, Tanabe K (1974) Ultrastructural changes associated with UV irradiation in the “germinal plasm” of Xenopus laevis. Dev Biol 36:155–168. https://doi.org/10.1016/0012-1606(74)90198-5
Iwamoto T, Hiraku Y, Oikawa S, Mizutani H, Kojima M, Kawanishi S (2004) DNA intrastrand cross-link at the 5′-GA-3′ sequence formed by busulfan and its role in the cytotoxic effect. Cancer Sci 95:454–458. https://doi.org/10.1111/j.1349-7006.2004.tb03231.x
Kawakami Y, Goto-Kazeto R, Saito T, Fujimoto T, Higaki S, Takahashi Y, Arai K, Yamaha E (2010) Generation of germ-line chimera zebrafish using primordial germ cells isolated from cultured blastomeres and cryopreserved embryoids. Int J Dev Biol 54:1491–1499. https://doi.org/10.1387/ijdb.093059yk
Kobayashi T, Takeuchi Y, Takeuchi T, Yoshizaki G (2007) Generation of viable fish from cryopreserved primordial germ cells. Mol Reprod Dev 74:207–213. https://doi.org/10.1002/mrd
Koprunner M, Thisse C, Thisse B, Raz E (2001) A zebrafish nanos-related gene is essential for the development of primordial germ cells. Genes Dev 15:2877–2885. https://doi.org/10.1101/gad.212401
Kurokawa H, Aoki Y, Nakamura S, Ebe Y, Kobayashi D, Tanaka M (2006) Time-lapse analysis reveals different modes of primordial germ cell migration in the medaka Oryzias latipes. Develop Growth Differ 48:209–221. https://doi.org/10.1111/j.1440-169X.2006.00858.x
Lacerda S, Batlouni S, Silva S, Homem C, França L (2006) Germ cells transplantation in fish: the Nile-tilapia model. Anim Reprod 3:146–159
Lacerda SM, Maria S, Costa GMJ, Da Silva Mde A, Almeida Campos-Junior PH, Segatelli TM, Peixoto MTD, Resende RR, De França LR (2013) Phenotypic characterization and in vitro propagation and transplantation of the Nile tilapia (Oreochromis niloticus) spermatogonial stem cells. Gen Comp Endocrinol 192:95–106. https://doi.org/10.1016/j.ygcen.2013.06.013
Leal MC, Cardoso ER, Nóbrega RH, Batlouni SR, Bogerd J, França LR, Schulz RW (2009) Histological and stereological evaluation of zebrafish (Danio rerio) spermatogenesis with an emphasis on spermatogonial generations. Biol Reprod 81:177–187. https://doi.org/10.1095/biolreprod.109.076299
Lee S, Iwasaki Y, Shikina S, Yoshizaki G (2013) Generation of functional eggs and sperm from cryopreserved whole testes. Proc Natl Acad Sci U S A 110:1640–1645. https://doi.org/10.1073/pnas.1218468110
Lee S, Seki S, Katayama N, Yoshizaki G (2015) Production of viable trout offspring derived from frozen whole fish. Sci Rep 5:16045. https://doi.org/10.1038/srep16045
Lin S, Long W, Chen J, Hopkins N (1992) Production of germ-line chimeras in zebrafish by cell transplants from genetically pigmented to albino embryos. Proc Natl Acad Sci U S A 89:4519–4523. https://doi.org/10.1073/pnas.89.10.4519
Linhartová Z, Saito T, Kašpar V, Rodina M, Prášková E, Hagihara S, Pšenička M (2015) Sterilization of sterlet Acipenser ruthenus by using knockdown agent, antisense morpholino oligonucleotide, against dead end gene. Theriogenology 84(7):1246–1255
Maxime V (2008) The physiology of triploid fish: current knowledge and comparisons with diploid fish. Fish Fish 9:67–78. https://doi.org/10.1111/j.1467-2979.2007.00269.x
Mishima Y (2012) Widespread roles of microRNAs during zebrafish development and beyond. Develop Growth Differ 54:55–65. https://doi.org/10.1111/j.1440-169X.2011.01306.x
Mishima Y, Giraldez AJ, Takeda Y, Fujiwara T, Sakamoto H, Schier AF, Inoue K (2006) Differential regulation of germline mRNAs in soma and germ cells by zebrafish miR-430. Curr Biol 16:2135–2142. https://doi.org/10.1016/j.cub.2006.08.086
Nagasawa K, Shikina S, Takeuchi Y, Yoshizaki G (2010) Lymphocyte antigen 75 (Ly75/CD205) is a surface marker on mitotic germ cells in rainbow trout. Biol Reprod 83:597–606. https://doi.org/10.1095/biolreprod.109.082081
Nagasawa K, Fernandes JMO, Yoshizaki G, Miwa M, Babiak I (2013) Identification and migration of primordial germ cells in Atlantic salmon, Salmo salar: characterization of vasa, dead end, and lymphocyte antigen 75 genes. Mol Reprod Dev 80:118–131. https://doi.org/10.1002/mrd.22142
Nóbrega RH, Greebe CD, van de Kant H, Bogerd J, de França LR, Schulz RW (2010) Spermatogonial stem cell niche and spermatogonial stem cell transplantation in zebrafish. PLoS One 5:1–16. https://doi.org/10.1371/journal.pone.0012808
Okutsu T, Suzuki K, Takeuchi Y, Takeuchi T, Yoshizaki G (2006) Testicular germ cells can colonize sexually undifferentiated embryonic gonad and produce functional eggs in fish. Proc Natl Acad Sci U S A 103:2725–2729. https://doi.org/10.1073/pnas.0509218103
Okutsu T, Shikina S, Kanno M, Takeuchi Y, Yoshizaki G (2007) Production of trout offspring from triploid salmon parents. Science 317(5844):1517
Olsen LC, Aasland R, Fjose A (1997) A vasa-like gene in zebrafish identifies putative primordial germ cells. Mech Dev 66:95–105. https://doi.org/10.1016/S0925-4773(97)00099-3
Omoto N, Maebayashi M, Adachi S, Arai K, Yamauchi K (2005) Sex ratios of triploids and gynogenetic diploids induced in the hybrid sturgeon, the bester (Huso huso female x Acipenser ruthenus male). Aquaculture 245:39–47. https://doi.org/10.1016/j.aquaculture.2004.12.004
Panda RP, Barman HK, Mohapatra C (2011) Isolation of enriched carp spermatogonial stem cells from Labeo rohita testis for in vitro propagation. Theriogenology 76:241–251. https://doi.org/10.1016/j.theriogenology.2011.01.031
Piferrer F, Beaumont A, Falguière JC, Flajšhans M, Haffray P, Colombo L (2009) Polyploid fish and shellfish: production, biology and applications to aquaculture for performance improvement and genetic containment. Aquaculture 293:125–156. https://doi.org/10.1016/j.aquaculture.2009.04.036
Pocherniaieva K, Sidova M, Havelka M, Saito T, Psenicka M, Sindelka R, Kaspar V (2018) Comparison of oocyte mRNA localization patterns in sterlet Acipenser ruthenus and African clawed frog Xenopus laevis. J Exp Zool B Mol Dev Evol 330(3):181–187
Pšenička M, Saito T, Linhartová Z, Gazo I (2015) Isolation and transplantation of sturgeon early-stage germ cells. Theriogenology 83:1085–1092. https://doi.org/10.1016/j.theriogenology.2014.12.010
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(2):119–122. https://doi.org/10.1016/j.cryobiol.2016.02.005
Saito T, Pšenička M (2015) Novel technique for visualizing primordial germ cells in sturgeons (Acipenser ruthenus, A. gueldenstaedtii, A. baerii, Huso huso). Biol Reprod 93(4):96. https://doi.org/10.1095/biolreprod.115.128314
Saito T, Fujimoto T, Maegawa S, Inoue K, Tanaka M, Arai K, Yamaha E (2006) Visualization of primordial germ cells in vivo using GFP-nos1 3′UTR mRNA. Int J Dev Biol 50:691–700. https://doi.org/10.1387/ijdb.062143ts
Saito T, Goto-Kazeto R, Arai K, Yamaha E (2008) Xenogenesis in teleost fish through generation of germ-line chimeras by single primordial germ cell transplantation. Biol Reprod 78:159–166. https://doi.org/10.1095/biolreprod.107.060038
Saito T, Goto-Kazeto R, Fujimoto T, Kawakami Y, Arai K, Yamaha E (2010) Inter-species transplantation and migration of primordial germ cells in cyprinid fish. Int J Dev Biol 54:1479–1484. https://doi.org/10.1387/ijdb.103111ts
Saito T, Goto-Kazeto R, Kawakami Y, Nomura K, Tanaka H, Adachi S, Arai K, Yamaha E (2011) The mechanism for primordial germ-cell migration is conserved between Japanese eel and zebrafish. PLoS One 6:1–8. https://doi.org/10.1371/journal.pone.0024460
Saito T, Pšenička M, Goto R, Adachi S, Inoue K, Arai K, Yamaha E (2014) The origin and migration of primordial germ cells in sturgeons. PLoS One 9(2):e86861. https://doi.org/10.1371/journal.pone.0086861
Seydoux G, Dunn MA (1997) Transcriptionally repressed germ cells lack a subpopulation of phosphorylated RNA polymerase II in early embryos of Caenorhabditis elegans and Drosophila melanogaster. Development 124:2191–2201
Seydoux G, Mello CC, Pettitt J, Wood WB, Priess JR, Fire A (1996) Repression of gene expression in the embryonic germ lineage of C. elegans. Nature 382:713–716. https://doi.org/10.1038/382713a0
Shikina S, Ihara S, Yoshizaki G (2008) Culture conditions for maintaining the survival and mitotic activity of rainbow trout transplantable type a spermatogonia. Mol Reprod Dev 75:529–537
Sutasurja LA, Nieuwkoop PD (1974) The induction of the primordial germ cells in the urodeles. Dev Genes Evol 175:199–220. https://doi.org/10.1007/BF00582092
Takeuchi Y, Yoshizaki G, Takeuchi T (2001) Production of germ-line chimeras in rainbow trout by blastomere transplantation. Mol Reprod Dev 59:380–389. https://doi.org/10.1002/mrd.1044
Takeuchi Y, Yoshizaki G, Takeuchi T (2003) Generation of live fry from intraperitoneally transplanted primordial germ cells in rainbow trout. Biol Reprod 69:1142–1149. https://doi.org/10.1095/biolreprod.103.017624
Takeuchi Y, Yoshizaki G, Takeuchi T (2004) Biotechnology: surrogate broodstock produces salmonids. Nature 430:629–630. https://doi.org/10.1038/430629a
Von Schönfeldt V, Krishnamurthy H, Foppiani L, Schlatt S (1999) Magnetic cell sorting is a fast and effective method of enriching viable spermatogonia from djungarian hamster, mouse, and marmoset monkey testes. Biol Reprod 61:582–589
Weidinger G, Stebler J, Slanchev K, Dumstrei K, Wise C, Lovell-Badge R, Thisse C, Thisse B, Raz E (2003) Dead end, a novel vertebrate germ plasm component, is required for zebrafish primordial germ cell migration and survival. Curr Biol 13:1429–1434. https://doi.org/10.1016/S0960-9822(03)00537-2
Whitington PM, Dixon KE (1975) Quantitative studies of germ plasm and germ cells during early embryogenesis of Xenopus laevis. J Embryol Exp Morphol 33:57–74
Williamson A, Lehmann R (1996) Germ cell development in Drosophila. Annu Rev Cell Dev Biol 12:365–391. https://doi.org/10.1146/annurev.cellbio.12.1.365
Wong T-T, Zohar Y (2015) Production of reproductively sterile fish by a non-transgenic gene silencing technology. Sci Rep 221:1–6. https://doi.org/10.1016/j.ygcen.2014.12.012
Wong T-T, Saito T, Crodian J, Collodi P (2011) Zebrafish germline chimeras produced by transplantation of ovarian germ cells into sterile host larvae. Biol Reprod 84:1190–1197. https://doi.org/10.1095/biolreprod.110.088427
Yamaha E, Kazama-Wakabayashi M, Otani S, Fujimoto T, Arai K (2001) Germ-line chimera by lower-part blastoderm transplantation between diploid goldfish and triploid crucian carp. Genetica 111:227–236. https://doi.org/10.1023/A:1013780423986
Yamaha E, Saito T, Goto-Kazeto R, Arai K (2007) Developmental biotechnology for aquaculture, with special reference to surrogate production in teleost fishes. J Sea Res 58:8–22. https://doi.org/10.1016/j.seares.2007.02.003
Yang X, Yue H, Ye H, Li C, Wei Q (2015) Identification of a germ cell marker gene, the dead end homologue, in Chinese sturgeon Acipenser sinensis. Gene 558:118–125. https://doi.org/10.1016/j.gene.2014.12.059
Yasui GS, Fujimoto T, Sakao S, Yamaha E, Arai K (2011) Production of loach (Misgurnus anguillicaudatus) germ-line chimera using transplantation of primordial germ cells isolated from cryopreserved blastomeres. J Anim Sci 89:2380–2388. https://doi.org/10.2527/jas.2010-3633
Yoon C, Kawakami K, Hopkins N (1997) Zebrafish vasa homologue RNA is localized to the cleavage planes of 2- and 4-cell-stage embryos and is expressed in the primordial germ cells. Development 124(16):3157–3165
Yoshizaki G, Ichikawa M, Hayashi M, Iwasaki Y, Miwa M, Shikina S, Okutsu T (2010) Sexual plasticity of ovarian germ cells in rainbow trout. Development 137:1227–1230. https://doi.org/10.1242/dev.051821
Yoshizaki G, Fujinuma K, Iwasaki Y, Okutsu T, Shikina S, Yazawa R, Takeuchi Y (2011) Spermatogonial transplantation in fish: a novel method for the preservation of genetic resources. Comp Biochem Physiol Part D Genomics Proteomics 6:55–63. https://doi.org/10.1016/j.cbd.2010.05.003
Acknowledgment
This study has been financially supported by the Ministry of Education, Youth and Sports of the Czech Republic-projects CENAKVA (LM2018099), project Biodiversity (CZ.02.1.01/0.0/0.0/16_025/0007370), and the Czech Science Foundation (grant number 16-02407Y).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Nature Singapore Pte Ltd.
About this chapter
Cite this chapter
Pšenička, M., Saito, T. (2020). Chapter 16 Specificity of Germ Cell Technologies in Sturgeons. In: Yoshida, M., Asturiano, J. (eds) Reproduction in Aquatic Animals. Springer, Singapore. https://doi.org/10.1007/978-981-15-2290-1_17
Download citation
DOI: https://doi.org/10.1007/978-981-15-2290-1_17
Published:
Publisher Name: Springer, Singapore
Print ISBN: 978-981-15-2289-5
Online ISBN: 978-981-15-2290-1
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)