Spermatogonial Stem Cell Niche and Spermatogonial Stem Cell Transplantation in Zebrafish

Nóbrega, Rafael Henrique; Greebe, Caaj Douwe; Kant, Henk van de; Bogerd, Jan; França, Luiz R.; Schulz, Rüdiger

doi:10.1371/journal.pone.0012808

Cited by 161 publications

(158 citation statements)

References 68 publications

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“…Testis tissue from transgenic fish was digested with 0.2% collagenase and 0.12% dispase as described previously (Nó brega et al 2010). The resulting cell suspension was immediately submitted to fluorescence-activated cell sorting (FACS) using an in Flux cell sorter (BD Bioscience, San Jose, CA, USA, www.bdbiosciences.com).…”

Section: Expression Of Cyp17a1 and Cyp19a1 Genes In Sorted Testicularmentioning

confidence: 99%

“…Then, Egfppositive and -negative cells were collected into tubes, centrifuged in PBS (pH 7.4) for 10 min at 100 g and immediately submitted to RNA extraction using RNAqueous -Micro Kit (Ambion, Austin, TX, USA, http://www. ambion.com) as described earlier (Nó brega et al 2010). After cDNA synthesis, the threshold cycle (Ct) values of two endogenous reference genes (elongation factor 1-a, ef1a; 18S rRNA) and of the target genes were determined by qPCR.…”

Section: Expression Of Cyp17a1 and Cyp19a1 Genes In Sorted Testicularmentioning

confidence: 99%

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Cyp17a1 and Cyp19a1 in the zebrafish testis are differentially affected by oestradiol

Hinfray¹,

Nóbrega²,

Caulier³

et al. 2013

Journal of Endocrinology

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International audienceOestrogens can affect expression of genes encoding steroidogenic enzymes in fish gonads. However, little information is available on their effects at the protein level. In this context, we first analysed the expression of key steroidogenic enzyme genes and proteins in zebrafish testis, paying attention also to other cell types than Leydig cells. Gene expression was analysed by quantitative PCR on fluorescence-activated cell-sorting fractions coupled or not to differential plating, while protein synthesis was studied by immunohistochemistry using specific antibodies against zebrafish Cyp17a1, Cyp19a1a and Cyp19a1b. Furthermore, we have evaluated the effect of oestrogen treatment (17 beta-oestradiol (E-2), 10 nM) on the localization of these enzymes after 7 and 14 days of in vivo exposure in order to study how oestrogen-mediated modulation of their expression is linked to oestrogen effects on spermatogenesis. The major outcomes of this study are that Leydig cells express Cyp17a1 and Cyp19a1a, while testicular germ cells express Cyp17a1 and both, Cyp19a1a and Cyp19a1b. As regards Cyp17a1, both protein and mRNA seem to be quantitatively dominating in Leydig cells. Moreover, E-2 exposure specifically affects only Leydig cell Cyp17a1 synthesis, preceding the disruption of spermatogenesis. The oestrogen-induced suppression of the androgen production capacity in Leydig cells is a major event in altering spermatogenesis, while germ cell steroidogenesis may have to be fuelled by precursors from Leydig cells. Further studies are needed to elucidate the functionality of steroidogenic enzymes in germ cells and their potential role in testicular physiology

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Section: Expression Of Cyp17a1 and Cyp19a1 Genes In Sorted Testicularmentioning

confidence: 99%

Section: Expression Of Cyp17a1 and Cyp19a1 Genes In Sorted Testicularmentioning

confidence: 99%

Cyp17a1 and Cyp19a1 in the zebrafish testis are differentially affected by oestradiol

Hinfray¹,

Nóbrega²,

Caulier³

et al. 2013

Journal of Endocrinology

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“…However, this in vitro differentiation assay for premeiotic germ cells cannot exclude the possibility that mitotic spermatogonia were already affected by the defective somatic cells and lost competency to progress through meiosis before culturing. Further experiments, such as germ cell transplantations (Kawasaki et al, 2010;Nóbrega et al, 2010), should be performed to elucidate in which cell types these genes are required for normal progression of spermatogenesis.…”

Section: In Vitro Differentiation Assay For Premeiotic Germ Cells Usimentioning

confidence: 99%

Isolation and cytogenetic characterization of zebrafish meiotic prophase I mutants

Saito

Siegfried

Nüsslein‐Volhard

et al. 2011

Developmental Dynamics

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We describe here the isolation and cytogenetic characterization of three meiotic prophase I mutants, denoted ietsugu (its), iesada (isa), and iemochi (imo), isolated by a novel N-ethyl-N-nitrosourea mutagenesis screen for adult zebrafish gonadogenesis. Histological examination and flow cytometry analysis of testes from these mutants showed that each contained neither spermatids nor sperm. Staining for Sycp3 and cleaved Caspase-3 and TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphosphate nick end-labeling) assay further revealed that its had defects at the onset of meiosis, and that isa and imo spermatocytes failed to progress past the zygotene stage with apoptosis occurring in the testicular somatic cells. Staining for phosphorylated histone H2AX showed that foci formation in leptotene spermatocytes was disrupted in isa and imo. Furthermore, in vitro differentiation experiments revealed the possibility that the defects and sterility associated with mutations were germ line autonomous. Our results thus indicate that each responsible gene is necessary for meiotic progression during spermatogenesis and for male fertility.

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“…They considered the mechanism of PGC migration to be highly conserved in fishes, and found that transplanting a single PGC was more efficient than transplanting blastomeres. Nobrega et al [107] transplanted spermatogonial stem cells of zebrafish into male and female recipients. From the results of donor-derived spermatogenesis and oogenesis, they demonstrated that germ cell differentiation could be influenced by the environment.…”

Section: Cell Engineering In Fish Reproductive Technologymentioning

confidence: 99%

Progress in studies of fish reproductive development regulation

Zhu

2012

Chin. Sci. Bull.

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Mechanisms of the animal reproductive development are an important research field in life sciences. The study of the reproductive development and regulatory mechanisms in fishes is important for elucidating the mechanisms of animal reproduction. This paper summarizes recent advances in the mechanisms of fish sex determination and differentiation, of fish gonad development and maturation, and of fish germ cell development, as well as the according regulating strategies. Fishes comprise an evolutionary stage that links invertebrates and higher vertebrates. They include diversiform species, and almost all vertebrate types of reproduction have been found in fishes. All these will lead to important advances in the regulatory mechanisms of animal reproduction by using fishes as model organisms. It will also enable novel fish breeding techniques when new controllable on-off strategies of reproduction and/or sex in fishes have been developed. fish, reproduction, regulation Citation:Chen J, Hu W, Zhu Z Y. Progress in studies of fish reproductive development regulation. Chin Sci Bull, 2013, 58: 716, doi: 10.1007/s11434-012-5577-1As low vertebrates, fishes are on an evolutionary stage linking invertebrates and higher vertebrates. There are as many as 28000 known fish species, which show large diversity. A range of sexual characteristics and reproductive strategies has been described for fishes, and almost all reproductive types of vertebrates have been found in fishes. The study of fish reproductive development regulation is important for elucidating mechanisms of animal reproductive development. Molecular breeding will reveal finer varieties when genes and regulatory networks associated with economically important traits (such as fast growth, disease resistance, cold tolerance and hypoxia tolerance) in fishes are discovered [1]. Reproduction also provides a precondition for culturing exclusive fish breeds and expanding fish populations. Breeding techniques such as sexual control and fertility control have played important roles in developing new fish varieties. This paper will review advances of fish reproductive development regulation, with the aim of providing hints for mechanisms of fish reproductive development, as well as providing theoretical guidance for techniques of reproductive regulation and developing highyielding, efficient, eco-friendly breeds.1 Sexual control of fishes Factors influencing sex determinationThe sex differentiation pattern and determination mechanism of fishes are very diverse. Various sexual phenotypes are all present in fishes, such as hermaphroditism, unisexuality, gonochorism and dual reproduction mode including parthenogenesis and sexual reproduction [2]. For those fishes in which sex chromosome systems exist, there are single chromosome systems, such as XX/XY, ZZ/ZW, XX/ XO and ZZ/ZO, and multiple chromosome systems, such as XX/XY 1 Y 2 , ZZ/ZW 1 W 2 , X 1 X 2 X 1 X 2 /X 1 X 2 Y, and X 1 X 2 X 1 X 2 / X 1 X 2 X 1 . Like other animals, fish sex differentiation is controlled by their gene...

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Spermatogonial Stem Cell Niche and Spermatogonial Stem Cell Transplantation in Zebrafish

Cited by 161 publications

References 68 publications

Cyp17a1 and Cyp19a1 in the zebrafish testis are differentially affected by oestradiol

Cyp17a1 and Cyp19a1 in the zebrafish testis are differentially affected by oestradiol

Isolation and cytogenetic characterization of zebrafish meiotic prophase I mutants

Progress in studies of fish reproductive development regulation

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