Sturgeons are among the most ancient linages of actinopterygians. At present, many sturgeon species are critically endangered. Surrogate production could be used as an affordable and a time-efficient method for endangered sturgeons. Our study established a method for identifying and isolating type A spermatogonia from different developmental stages of testes using flow cytometric cell sorting (FCM). Flow cytometric analysis of a whole testicular cell suspension showed several well-distinguished cell populations formed according to different values of light scatter parameters. FCM of these different cell populations was performed directly on glass slides for further immunocytochemistry to identify germ cells. Results showed that the cell population in gate P1 on a flow cytometry plot (with high forward scatter and high side scatter parameter values) contains the highest amount of type A spermatogonia. The sorted cell populations were characterized by expression profiles of 10 germ cell specific genes. The result confirmed that setting up for the P1 gate could precisely sort type A spermatogonia in all tested testicular developmental stages. The P2 gate, which was with lower forward scatter and side scatter values mostly, contained type B spermatogonia at a later maturing stage. Moreover, expressions of plzf, dnd, boule, and kitr were significantly higher in type A spermatogonia than in later developed germ cells. In addition, plzf was firstly found as a reliable marker to identify type A spermatogonia, which filled the gap of identification of spermatogonial stem cells in sterlet. It is expected to increase the efficiency of germ stem cell culture and transplantation with plzf identification. Our study thus first addressed a phenotypic characterization of a pure type A spermatogonia population in sterlet. FCM strategy can improve the production of sturgeons with surrogate broodstock and further the analysis of the cellular and molecular mechanisms of sturgeon germ cell development.

Biology Center of the Czech Academy of Sciences Institute of Entomology České Budějovice Czechia

Department of Pharmacology C_DAT University Medicine Greifswald Greifswald Germany

Imaging Methods Core Facility at BIOCEV Operated by Faculty of Science Charles University Prague Vestec Czechia

Laboratory of Gene Expression Institute of Biotechnology of the Czech Academy of Sciences Vestec Czechia

South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses Faculty of Fisheries and Protection of Waters Research Institute of Fish Culture and Hydrobiology University of South Bohemia in České Budějovice Vodňany Czechia

University of South Bohemia Faculty of Science České Budějovice Czechia

Zobrazit více v PubMed

Aramaki S., Kato T., Soh T., Yamauchi N., Hattori M. A. (2008). Molecular Cloning and Expression of Dead End Homologue in Chicken Germ Cells. Cell Tissue Res 330, 45–52. 10.1093/biolreprod/78.s1.298d PubMed DOI

Baloch A. R., Franěk R., Tichopád T., Fučíková M., Rodina M., Pšenička M. (2019a). Dnd1 Knockout in Sturgeons by CRISPR/Cas9 Generates Germ Cell Free Host for Surrogate Production. Animals 9, 174. 10.3390/ani9040174 PubMed DOI PMC

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 Testis1. Biol. Reprod. 90, 1–6. 10.1095/biolreprod.113.116392 PubMed DOI

Bemis W. E., Findeis E. K., Grande L. (1997). An Overview of Acipenseriformes. Environ. Biol. Fishes 48, 25–71. 10.1023/A:1007370213924 DOI

Bemis W. E., Kynard B. (1997). Sturgeon Rivers: an Introduction to Acipenseriform Biogeography and Life History. Environ. Biol. Fishes 48, 167–183. 10.1023/a:1007312524792 DOI

Berbejillo J., Martinez-Bengochea A., Bedo G., Brunet F., Volff J.-N., Vizziano-Cantonnet D. (2012). Expression and Phylogeny of Candidate Genes for Sex Differentiation in a Primitive Fish Species, the Siberian sturgeon, Acipenser baerii . Mol. Reprod. Dev. 79, 504–516. 10.1002/mrd.22053 PubMed DOI

Bronzi P., Rosenthal H. (2014). Present and Future sturgeon and Caviar Production and Marketing: A Global Market Overview. J. Appl. Ichthyol. 30, 1536–1546. 10.1111/jai.12628 DOI

Buaas F. W., Kirsh A. L., Sharma M., McLean D. J., Morris J. L., Griswold M. D., et al. (2004). Plzf Is Required in Adult Male Germ Cells for Stem Cell Self-Renewal. Nat. Genet. 36, 647–652. 10.1038/ng1366 PubMed DOI

De Rooij D. G., Russell L. D. (2000). All You Wanted to Know about Spermatogonia but Were Afraid to Ask. J. Androl. 21, 776–798. PubMed

Dolci S., Williams D. E., Ernst M. K., Resnick J. L., Brannan C. I., Lock L. F., et al. (1991). Requirement for Mast Cell Growth Factor for Primordial Germ Cell Survival in Culture. Nature 352, 809–811. 10.1038/352809a0 PubMed DOI

Eberhart C. G., Maines J. Z., Wasserman S. A. (1996). Meiotic Cell Cycle Requirement for a Fly Homologue of Human Deleted in Azoospermia. Nature 381, 783–785. 10.1038/381783a0 PubMed DOI

Fajkowska M., Głowacka D. K., Adamek-Urbańska D., Ostaszewska T., Krajnik K. A., Rzepkowska M. (2019). Sex-related Gene Expression Profiles in Various Tissues of Juvenile Russian sturgeon (Acipenser gueldenstaedtii). Aquaculture 500, 532–539. 10.1016/j.aquaculture.2018.10.066 DOI

Fajkowska M., Ostaszewska T., Rzepkowska M. (2020). Review: Molecular Mechanisms of Sex Differentiation in Sturgeons. Rev. Aquacult 12, 1003–1027. 10.1111/raq.12369 DOI

Fajkowska M., Rzepkowska M., Adamek D., Ostaszewska T., Szczepkowski M. (2016). Expression of Dmrt1 and Vtg Genes during Gonad Formation, Differentiation and Early Maturation in Cultured Russian sturgeon Acipenser gueldenstaedtii . J. Fish. Biol. 89, 1441–1449. 10.1111/jfb.12992 PubMed DOI

Franěk R., Marinović Z., Lujić J., Urbányi B., Fučíková M., Kašpar V., et al. (2019). Cryopreservation and Transplantation of Common Carp Spermatogonia. PLoS One 14, e0205481. 10.1371/journal.pone.0205481 PubMed DOI PMC

Franěk R., Kašpar V., Shah M. A., Gela D., Pšenička M. (2021). Production of Common Carp Donor-Derived Offspring from Goldfish Surrogate Broodstock. Aquaculture 534, 736252. 10.1016/j.aquaculture.2020.736252 DOI

Gautier A., Bosseboeuf A., Auvray P., Sourdaine P. (2014). Maintenance of Potential Spermatogonial Stem Cells In Vitro by GDNF Treatment in a Chondrichthyan Model (Scyliorhinus canicula L.)1. Biol. Reprod. 91. 10.1095/biolreprod.113.116020 PubMed DOI

Hagihara S., Yamashita R., Yamamoto S., Ishihara M., Abe T., Ijiri S., et al. (2014). Identification of Genes Involved in Gonadal Sex Differentiation and the Dimorphic Expression Pattern in Undifferentiated Gonads of Russian sturgeon Acipenser gueldenstaedtii Brandt & Ratzeburg, 1833. J. Appl. Ichthyol. 30, 1557–1564. 10.1111/jai.12588 DOI

Hayashi M., Sato M., Nagasaka Y., Sadaie S., Kobayashi S., Yoshizaki G. (2014). Enrichment of Spermatogonial Stem Cells Using Side Population in Teleost1. Biol. Reprod. 91, 1–8. 10.1095/biolreprod.113.114140 PubMed DOI

Higuchi K., Takeuchi Y., Miwa M., Yamamoto Y., Tsunemoto K., Yoshizaki G. (2011). Colonization, Proliferation, and Survival of Intraperitoneally Transplanted Yellowtail Seriola quinqueradiata Spermatogonia in Nibe Croaker Nibea mitsukurii Recipient. Fish. Sci. 77, 69–77. 10.1007/s12562-010-0314-7 DOI

Horvay K., Claußen M., Landgrebe J., Pieler T. (2006). Clau?EnXenopus Dead End mRNA Is a Localized Maternal Determinant that Serves a Conserved Function in Germ Cell Development. Developmental Biol. 291, 1–11. 10.1016/j.ydbio.2005.06.013 PubMed DOI

Ichida K., Kawamura W., Miwa M., Iwasaki Y., Kubokawa T., Hayashi M., et al. (2019). Specific Visualization of Live Type A Spermatogonia of Pacific Bluefin Tuna Using Fluorescent Dye-Conjugated Antibodies†. Biol. Reprod. 100, 1637–1647. 10.1093/biolre/ioz047 PubMed DOI

Ichida K., Kise K., Morita T., Yazawa R., Takeuchi Y., Yoshizaki G. (2017). Flow-cytometric Enrichment of Pacific Bluefin Tuna Type A Spermatogonia Based on Light-Scattering Properties. Theriogenology 101, 91–98. 10.1016/j.theriogenology.2017.06.022 PubMed DOI

Inoue J. G., Miya M., Venkatesh B., Nishida M. (2005). The Mitochondrial Genome of Indonesian Coelacanth Latimeria Menadoensis (Sarcopterygii: Coelacanthiformes) and Divergence Time Estimation between the Two Coelacanths. Gene 349, 227–235. 10.1016/J.GENE.2005.01.008 PubMed DOI

Iwasaki-Takahashi Y., Shikina S., Watanabe M., Banba A., Yagisawa M., Takahashi K., et al. (2020). Production of Functional Eggs and Sperm from In Vitro-expanded Type A Spermatogonia in Rainbow trout. Commun. Biol. 3, 308. 10.1038/s42003-020-1025-y PubMed DOI PMC

Kanatsu-Shinohara M., Morimoto H., Shinohara T. (2012). Enrichment of Mouse Spermatogonial Stem Cells by Melanoma Cell Adhesion Molecule Expression. Biol. Reprod. 87, 139. 10.1095/biolreprod.112.103861 PubMed DOI

Kise K., Yoshikawa H., Sato M., Tashiro M., Yazawa R., Nagasaka Y., et al. (2012). Flow-Cytometric Isolation and Enrichment of Teleost Type A Spermatogonia Based on Light-Scattering Properties1. Biol. Reprod. 86. 10.1095/biolreprod.111.093161 PubMed DOI

Kokkinaki M., Djourabtchi A., Golestaneh N. (2011). Long-term Culture of Human SSEA-4 Positive Spermatogonial Stem Cells (SSCs). J. Stem Cell Res. Ther. 01. 10.4172/2157-7633.S2-003 PubMed DOI PMC

Lacerda S. M. d. S. N., Costa G. M. J., de França L. R. (2014). Biology and Identity of Fish Spermatogonial Stem Cell. Gen. Comp. Endocrinol. 207, 56–65. 10.1016/J.YGCEN.2014.06.018 PubMed DOI

Lacerda S. M. S. N., Batlouni S. R., Costa G. M. J., Segatelli T. M., Quirino B. R., Queiroz B. M., et al. (2010). A New and Fast Technique to Generate Offspring after Germ Cells Transplantation in Adult Fish: The Nile tilapia (Oreochromis niloticus) Model. PLoS One 5, e10740. 10.1371/journal.pone.0010740 PubMed DOI PMC

Lacerda S. M. S. N., Martinez E. R. M., Mura I. L. D. D., Doretto L. B., Costa G. M. J., Silva M. A., et al. (2019). Duration of Spermatogenesis and Identification of Spermatogonial Stem Cell Markers in a Neotropical Catfish, Jundiá (Rhamdia quelen). Gen. Comp. Endocrinol. 273, 249–259. 10.1016/j.ygcen.2018.10.018 PubMed DOI

Linhartová Z., Rodina M., Guralp H., Gazo I., Saito T., Pšenička M. (2014). Pšenička M.: Isolation and Cryopreservation of Early Stages of Germ Cells of Tench (Tinca tinca). Czech J. Anim. Sci. 59, 381–390. 10.17221/7589-cjas DOI

Linhartová Z., Saito T., Kašpar V., Rodina M., Prášková E., Hagihara S., et al. (2015). Sterilization of Sterlet Acipenser ruthenus by Using Knockdown Agent, Antisense Morpholino Oligonucleotide, against Dead End Gene. Theriogenology 84, 1246–1255. 10.1016/j.theriogenology.2015.07.003 PubMed DOI

Liu L., Hong N., Xu H., Li M., Yan Y., Purwanti Y., et al. (2009). Medaka Dead End Encodes a Cytoplasmic Protein and Identifies Embryonic and Adult Germ Cells. Gene Expr. Patterns 9, 541–548. 10.1016/j.gep.2009.06.008 PubMed DOI

Lujić J., Marinović Z., Bajec S. S., Djurdjevič I., Urbányi B., Horváth Á. (2018). Interspecific Germ Cell Transplantation: a New Light in the Conservation of Valuable Balkan trout Genetic Resources? Fish. Physiol. Biochem. 44, 1487–1498. 10.1007/s10695-018-0510-4 PubMed DOI

Maouche A., Curran E., Goupil A.-S., Sambroni E., Bellaiche J., Le Gac F., et al. (2018). New Insights into the Evolution, Hormonal Regulation, and Spatiotemporal Expression Profiles of Genes Involved in the Gfra1/Gdnf and Kit/Kitlg Regulatory Pathways in Rainbow trout Testis. Fish. Physiol. Biochem. 44, 1599–1616. 10.1007/s10695-018-0547-4 PubMed DOI

Mohapatra C., Barman H. K. (2014). Identification of Promoter within the First Intron of Plzf Gene Expressed in Carp Spermatogonial Stem Cells. Mol. Biol. Rep. 41, 6433–6440. 10.1007/s11033-014-3525-7 PubMed DOI

Morita T., Morishima K., Miwa M., Kumakura N., Kudo S., Ichida K., et al. (2015). Functional Sperm of the Yellowtail (Seriola quinqueradiata) Were Produced in the Small-Bodied Surrogate, jack Mackerel (Trachurus Japonicus). Mar. Biotechnol. 17, 644–654. 10.1007/s10126-015-9657-5 PubMed DOI

Nagasawa K., Fernandes J. M. O., 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. 10.1002/mrd.22142 PubMed DOI PMC

Nayak S., Ferosekhan S., Sahoo S. K., Sundaray J. K., Jayasankar P., Barman H. K. (2016). Production of fertile Sperm from In Vitro Propagating Enriched Spermatogonial Stem Cells of Farmed Catfish, Clarias batrachus . Zygote 24, 814–824. 10.1017/S0967199416000149 PubMed DOI

Okutsu T., Shikina S., Kanno M., Takeuchi Y., Yoshizaki G. (2007). Production of trout Offspring from Triploid salmon Parents. Science 317, 1517. 10.1126/science.1145626 PubMed DOI

Ozaki Y., Saito K., Shinya M., Kawasaki T., Sakai N. (2011). Evaluation of Sycp3, Plzf and Cyclin B3 Expression and Suitability as Spermatogonia and Spermatocyte Markers in Zebrafish. Gene Expr. Patterns 11, 309–315. 10.1016/j.gep.2011.03.002 PubMed DOI

Panda R. P., Barman H. K., Mohapatra C. (2011). Isolation of Enriched Carp Spermatogonial Stem Cells from Labeo Rohita Testis for In Vitro Propagation. Theriogenology 76, 241–251. 10.1016/j.theriogenology.2011.01.031 PubMed DOI

Pšenička M., Saito T., Linhartová Z., Gazo I. (2015). Isolation and Transplantation of sturgeon Early-Stage Germ Cells. Theriogenology 83, 1085–1092. 10.1016/J.THERIOGENOLOGY.2014.12.010 PubMed DOI

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, 119–122. 10.1016/j.cryobiol.2016.02.005 PubMed DOI

Ramirez J.-M., Bai Q., Péquignot M., Becker F., Kassambara A., Bouin A., et al. (2013). Side Scatter Intensity Is Highly Heterogeneous in Undifferentiated Pluripotent Stem Cells and Predicts Clonogenic Self-Renewal. Stem Cell Development 22, 1851–1860. 10.1089/scd.2012.0658 PubMed DOI PMC

Rolland A. D., Lareyre J.-J., Goupil A.-S., Montfort J., Ricordel M.-J., Esquerré D., et al. (2009). Expression Profiling of Rainbow trout Testis Development Identifies Evolutionary Conserved Genes Involved in Spermatogenesis. BMC Genomics 10. 10.1186/1471-2164-10-546 PubMed DOI PMC

Runyan C., Schaible K., Molyneaux K., Wang Z., Levin L., Wylie C. (2006). Steel Factor Controls Midline Cell Death of Primordial Germ Cells and Is Essential for Their normal Proliferation and Migration. Development 133 (24), 4861–4869. 10.1242/dev.02688 PubMed DOI

Santos Nassif Lacerda S. M., Costa G. M. J., da Silva M. d. A., Almeida Campos-Junior P. H., Segatelli T. M., Peixoto M. T. D., et al. (2013). Phenotypic Characterization and In Vitro Propagation and Transplantation of the Nile tilapia (Oreochromis niloticus) Spermatogonial Stem Cells. Gen. Comp. Endocrinol. 192, 95–106. 10.1016/J.YGCEN.2013.06.013 PubMed DOI

Satoh R., Bando H., Sakai N., Kotani T., Yamashita M. (2019). Function of Leukaemia Inhibitory Factor in Spermatogenesis of a Teleost Fish, the Medaka Oryzias latipes . Zygote 27, 423–431. 10.1017/S0967199419000558 PubMed DOI

Schulz R. W., de França L. R., Lareyre J.-J., LeGac F., Chiarini-Garcia H., Nobrega R. H., et al. (2010). Spermatogenesis in Fish. Gen. Comp. Endocrinol. 165, 390–411. 10.1016/j.ygcen.2009.02.013 PubMed DOI

Schulz R. W., Menting S., Bogerd J., França L. R., Vilela D. A. R., Godinho H. P. (2005). Sertoli Cell Proliferation in the Adult Testis-Evidence from Two Fish Species Belonging to Different Orders1. Biol. Reprod. 73, 891–898. 10.1095/biolreprod.105.039891 PubMed DOI

Shang M., Su B., Lipke E. A., Perera D. A., Li C., Qin Z., et al. (2015). Spermatogonial Stem Cells Specific Marker Identification in Channel Catfish, Ictalurus punctatus and Blue Catfish, I. furcatus . Fish. Physiol. Biochem. 41, 1545–1556. 10.1007/s10695-015-0106-1 PubMed DOI

Shang M., Su B., Perera D. A., Alsaqufi A., Lipke E. A., Cek S., et al. (2018). Testicular Germ Line Cell Identification, Isolation, and Transplantation in Two North American Catfish Species. Fish. Physiol. Biochem. 44, 717–733. 10.1007/s10695-018-0467-3 PubMed DOI

Takeuchi Y., Yoshizaki G., Takeuchi T. (2004). Surrogate Broodstock Produces Salmonids. Nature 430, 629–630. 10.1038/430629a PubMed DOI

Valli H., Sukhwani M., Dovey S. L., Peters K. A., Donohue J., Castro C. A., et al. (2014). Fluorescence- and Magnetic-Activated Cell Sorting Strategies to Isolate and Enrich Human Spermatogonial Stem Cells. Fertil. Sterility 102, 566–580. 10.1016/J.FERTNSTERT.2014.04.036 PubMed DOI PMC

Vizziano-Cantonnet D., Lasalle A., Di Landro S., Klopp C., Genthon C. (2018). De Novo transcriptome Analysis to Search for Sex-Differentiation Genes in the Siberian sturgeon. Gen. Comp. Endocrinol. 268, 96–109. 10.1016/j.ygcen.2018.08.007 PubMed DOI

Wang W., Zhu H., Dong Y., Dong T., Tian Z., Hu H. (2019). Identification and Dimorphic Expression of Sex-Related Genes during Gonadal Differentiation in Sterlet Acipenser ruthenus, a Primitive Fish Species. Aquaculture 500, 178–187. 10.1016/j.aquaculture.2018.10.001 PubMed DOI

Wang W., Zhu H., Dong Y., Tian Z., Dong T., Hu H., et al. (2017). Dimorphic Expression of Sex-Related Genes in Different Gonadal Development Stages of Sterlet, Acipenser ruthenus, a Primitive Fish Species. Fish. Physiol. Biochem. 43, 1557–1569. 10.1007/s10695-017-0392-x PubMed DOI

Weidinger G., Stebler J., Slanchev K., Dumstrei K., Wise C., Lovell-Badge R., et al. (2003). Dead End, a Novel Vertebrate Germ Plasm Component, Is Required for Zebrafish Primordial Germ Cell Migration and Survival. Curr. Biol. 13, 1429–1434. 10.1016/s0960-9822(03)00537-2 PubMed DOI

Wong T.-T., Saito T., Crodian J., Collodi P. (2011). Zebrafish Germline Chimeras Produced by Transplantation of Ovarian Germ Cells into Sterile Host Larvae1. Biol. Reprod. 84, 1190–1197. 10.1095/biolreprod.110.088427 PubMed DOI PMC

Xie X., Li P., Pšenička M., Ye H., Steinbach C., Li C., et al. (2019). Optimization of In Vitro Culture Conditions of sturgeon Germ Cells for Purpose of Surrogate Production. Animals 9, 106. 10.3390/ani9030106 PubMed DOI PMC

Xie X., Nóbrega R., Pšenička M. (2020). Spermatogonial Stem Cells in Fish: Characterization, Isolation, Enrichment, and Recent Advances of In Vitro Culture Systems. Biomolecules 10, 644. 10.3390/biom10040644 PubMed DOI PMC

Xu E. Y., Moore F. L., Pera R. A. R. (2001). A Gene Family Required for Human Germ Cell Development Evolved from an Ancient Meiotic Gene Conserved in Metazoans. Proc. Natl. Acad. Sci. 98, 7414–7419. 10.1073/pnas.131090498 PubMed DOI PMC

Xu H., Li Z., Li M., Wang L., Hong Y., Laura R. (2009). Boule Is Present in Fish and Bisexually Expressed in Adult and Embryonic Germ Cells of Medaka. PLoS One 4, e6097. 10.1371/journal.pone.0006097 PubMed DOI PMC

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. 10.1016/j.gene.2014.12.059 PubMed DOI

Ye H., Li C.-J., Yue H.-M., Du H., Yang X.-G., Yoshino T., et al. (2017). Establishment of Intraperitoneal Germ Cell Transplantation for Critically Endangered Chinese sturgeon Acipenser Sinensis. Theriogenology 94, 37–47. 10.1016/j.theriogenology.2017.02.009 PubMed DOI

Ye H., Li C.-J., Yue H.-M., Yang X.-G., Wei Q.-W. (2015). Differential Expression of Fertility Genes Boule and Dazl in Chinese sturgeon (Acipenser Sinensis), a Basal Fish. Cell Tissue Res 360, 413–425. 10.1007/s00441-014-2095-2 PubMed DOI

Ye H., Zhou C., Yue H., Wu M., Ruan R., Du H., et al. (2021). Cryopreservation of Germline Stem Cells in American Paddlefish (Polyodon spathula). Anim. Reprod. Sci. 224, 106667. 10.1016/j.anireprosci.2020.106667 PubMed DOI

Yoshikawa H., Takeuchi Y., Ino Y., Wang J., Iwata G., Kabeya N., et al. (2017). Efficient Production of Donor-Derived Gametes from Triploid Recipients Following Intra-peritoneal Germ Cell Transplantation into a marine Teleost, Nibe Croaker ( Nibea mitsukurii ). Aquaculture 478, 35–47. 10.1016/j.aquaculture.2016.05.011 DOI

Yoshizaki G., Okutsu T., Morita T., Terasawa M., Yazawa R., Takeuchi Y. (2012). Biological Characteristics of Fish Germ Cells and Their Application to Developmental Biotechnology. Reprod. Domest. Anim. 47, 187–192. 10.1111/j.1439-0531.2012.02074.x PubMed DOI

Youngren K. K., Coveney D., Peng X., Bhattacharya C., Schmidt L. S., Nickerson M. L., et al. (2005). The Ter Mutation in the Dead End Gene Causes Germ Cell Loss and Testicular Germ Cell Tumours. Nature 435, 360–364. 10.1038/nature03595 PubMed DOI PMC

Yue H., Li C., Du H., Zhang S., Wei Q. (2015). Sequencing and De Novo Assembly of the Gonadal Transcriptome of the Endangered Chinese sturgeon (Acipenser Sinensis). PLoS One 10, e0127332–22. 10.1371/journal.pone.0127332 PubMed DOI PMC

Zhang F., Hao Y., Li X., Li Y., Ye D., Zhang R., et al. (2021). Surrogate Production of Genome-Edited Sperm from a Different Subfamily by Spermatogonial Stem Cell Transplantation. Sci. China Life Sci. 1–19. 10.1007/s11427-021-1989-9 PubMed DOI

Zhang F., Li X., He M., Ye D., Xiong F., Amin G., et al. (2020a). Efficient Generation of Zebrafish Maternal-Zygotic Mutants through Transplantation of Ectopically Induced and Cas9/gRNA Targeted Primordial Germ Cells. J. Genet. Genomics 47, 37–47. 10.1016/j.jgg.2019.12.004 PubMed DOI

Zhang X., Zhou J., Li L., Huang W., Ahmad H. I., Li H., et al. (2020b). Full-length Transcriptome Sequencing and Comparative Transcriptomic Analysis to Uncover Genes Involved in Early Gametogenesis in the Gonads of Amur sturgeon (Acipenser Schrenckii). Front. Zool. 17, 1–21. 10.1186/s12983-020-00355-z PubMed DOI PMC

Zhuang P., Wei Q., He X., Cen Y. (1997). Biology and Life History of Dabry’s sturgeon, Acipenser Dabryanus, in the Yangtze River. Environ. Biol. Fishes 48, 257–264. 10.1023/a:1007399729080 DOI