17β-Trenbolone binds to androgen receptor, decreases number of primordial germ cells, modulates expression of genes related to sexual differentiation, and affects sexual differentiation in zebrafish (Danio rerio)

2021 ◽  
pp. 150959
Author(s):  
Hua Tian ◽  
Rui Liu ◽  
Suqiu Zhang ◽  
Shuhui Wei ◽  
Wei Wang ◽  
...  
Keyword(s):  
Androgen Receptor ◽  
Germ Cells ◽  
Danio Rerio ◽  
Sexual Differentiation ◽  
Primordial Germ Cells ◽  
Expression Of Genes

Effects of ten–eleven translocation 1 (Tet1) on DNA methylation and gene expression in chicken primordial germ cells

2019 ◽  
Vol 31 (3) ◽  
pp. 509 ◽  
Author(s):  
Minli Yu ◽  
Dongfeng Li ◽  
Wanyan Cao ◽  
Xiaolu Chen ◽  
Wenxing Du
Keyword(s):  
Gene Expression ◽  
Dna Methylation ◽  
Germ Cells ◽  
Dna Methyltransferase ◽  
Primordial Germ Cells ◽  
Dna Demethylation ◽  
Caveolin 1 ◽  
Expression Of Genes ◽  
Deleted In Azoospermia

Ten–eleven translocation 1 (Tet1) is involved in DNA demethylation in primordial germ cells (PGCs); however, the precise regulatory mechanism remains unclear. In the present study the dynamics of 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) in developing PGCs and the role of Tet1 in PGC demethylation were analysed. Results show that 5mC levels dropped significantly after embryonic Day 4 (E4) and 5hmC levels increased reaching a peak at E5–E5.5. Interestingly, TET1 protein was highly expressed during E5 to E5.5, which showed a consistent trend with 5hmC. The expression of pluripotency-associated genes (Nanog, PouV and SRY-box 2 (Sox2)) and germ cell-specific genes (caveolin 1 (Cav1), piwi-like RNA-mediated gene silencing 1 (Piwi1) and deleted in azoospermia-like (Dazl)) was upregulated after E5, whereas the expression of genes from the DNA methyltransferase family was decreased. Moreover, the Dazl gene was highly methylated in early PGCs and then gradually hypomethylated. Knockdown of Tet1 showed impaired survival and proliferation of PGCs, as well as increased 5mC levels and reduced 5hmC levels. Further analysis showed that knockdown of Tet1 led to elevated DNA methylation levels of Dazl and downregulated gene expression including Dazl. Thus, this study reveals the dynamic epigenetic reprogramming of chicken PGCs invivo and the molecular mechanism of Tet1 in regulating genomic DNA demethylation and hypomethylation of Dazl during PGC development.

scholarly journals Production of fertile zebrafish (Danio rerio) possessing germ cells (gametes) originated from primordial germ cells recovered from vitrified embryos

Reproduction ◽  
2010 ◽  
Vol 139 (4) ◽  
pp. 733-740 ◽  
Author(s):  
Shogo Higaki ◽  
Yoshiki Eto ◽  
Yutaka Kawakami ◽  
Etsuro Yamaha ◽  
Noriko Kagawa ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Ethylene Glycol ◽  
Germ Cells ◽  
Danio Rerio ◽  
Recovery Rate ◽  
Fluorescent Protein ◽  
Primordial Germ Cells ◽  
Ice Formation ◽  
Mature Fish ◽  
Green Fluorescent

This study aimed to produce fertile zebrafish (Danio rerio) possessing germ cells (gametes) that originated from cryopreserved primordial germ cells (PGCs). First, to improve the vitrification procedure of PGCs in segmentation stage embryos, dechorionated yolk-intact and yolk-removed embryos, the PGCs of which were labeled with green fluorescent protein, were cooled rapidly after serial exposures to equilibration solution (ES) and vitrification solution (VS), which contained ethylene glycol, DMSO, and sucrose. Yolk removal well prevented ice formation in the embryos during cooling and improved the viability of cryopreserved PGCs. The maximum recovery rate of live PGCs in the yolk-removed embryos vitrified after optimum exposure to ES and VS was estimated to be about 90%, and about 50% of the live PGCs showed pseudopodial movement. Next, to elucidate the ability of cryopreserved PGCs to differentiate into functional gametes, PGCs recovered from the yolk-removed embryos (striped-type) that were vitrified under the optimum exposure to ES and VS were transplanted individually into 218 sterilized recipient blastulae (golden-type). Two days after the transplantation, 7.5% (14/187) of morphologically normal embryos had PGC(s) in the genital ridges. Six (5 males and 1 female) of the 14 recipient embryos developed into mature fish and generated progeny with characteristics inherited from PGC donors. In conclusion, we demonstrated the successful cryopreservation of PGCs by vitrification of yolk-removed embryos and the production of fertile zebrafish possessing germ cells that originated from the PGCs in vitrified embryos.

scholarly journals SWI/SNF chromatin remodeling complex is required for initiation of sex-dependent differentiation in mouse germline

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Toshiaki Ito ◽  
Atsuki Osada ◽  
Masami Ohta ◽  
Kana Yokota ◽  
Akira Nishiyama ◽  
...  
Keyword(s):  
Chromatin Remodeling ◽  
Germ Cells ◽  
De Novo ◽  
Primordial Germ Cells ◽  
Strand Break ◽  
Conditional Knockout ◽  
Dna Hypomethylation ◽  
Aberrant Expression ◽  
Chromatin Remodeling Complex ◽  
Expression Of Genes

AbstractSexual reproduction involves the creation of sex-dependent gametes, oocytes and sperm. In mammals, sexually dimorphic differentiation commences in the primordial germ cells (PGCs) in embryonic gonads; PGCs in ovaries and testes differentiate into meiotic primary oocytes and mitotically quiescent prospermatogonia, respectively. Here, we show that the transition from PGCs to sex-specific germ cells was abrogated in conditional knockout mice carrying a null mutation of Smarcb1 (also known as Snf5) gene, which encodes a core subunit of the SWI/SNF chromatin remodeling complex. In female mutant mice, failure to upregulate meiosis-related genes resulted in impaired meiotic entry and progression, including defects in synapsis formation and DNA double strand break repair. Mutant male mice exhibited delayed mitotic arrest and DNA hypomethylation in retrotransposons and imprinted genes, resulting from aberrant expression of genes related to growth and de novo DNA methylation. Collectively, our results demonstrate that the SWI/SNF complex is required for transcriptional reprogramming in the initiation of sex-dependent differentiation of germ cells.

Cryopreservation of zebrafish (Danio rerio) primordial germ cells by vitrification of yolk-intact and yolk-depleted embryos using various cryoprotectant solutions

Cryobiology ◽  
2013 ◽  
Vol 67 (3) ◽  
pp. 374-382 ◽  
Author(s):  
Shogo Higaki ◽  
Yutaka Kawakami ◽  
Yoshiki Eto ◽  
Etsuro Yamaha ◽  
Masashi Nagano ◽  
...  
Keyword(s):  
Germ Cells ◽  
Danio Rerio ◽  
Primordial Germ Cells

scholarly journals A Drosophila toolkit for defining gene function in spermatogenesis

Reproduction ◽  
2017 ◽  
Vol 153 (4) ◽  
pp. R121-R132 ◽  
Author(s):  
N A Siddall ◽  
G R Hime
Keyword(s):  
Germ Cells ◽  
Gene Function ◽  
Primordial Germ Cells ◽  
Rapid Assessment ◽  
Model Organism ◽  
Time Frame ◽  
Individual Organism ◽  
Expression Of Genes ◽  
X Irradiation ◽  
Human Spermatogenesis

Expression profiling and genomic sequencing methods enable the accumulation of vast quantities of data that relate to the expression of genes during the maturation of male germ cells from primordial germ cells to spermatozoa and potential mutations that underlie male infertility. However, the determination of gene function in specific aspects of spermatogenesis or linking abnormal gene function with infertility remain rate limiting, as even in an era of CRISPR analysis of gene function in mammalian models, this still requires considerable resources and time. Comparative developmental biology studies have shown the remarkable conservation of spermatogenic developmental processes from insects to vertebrates and provide an avenue of rapid assessment of gene function to inform the potential roles of specific genes in rodent and human spermatogenesis. The vinegar fly, Drosophila melanogaster, has been used as a model organism for developmental genetic studies for over one hundred years, and research with this organism produced seminal findings such as the association of genes with chromosomes, the chromosomal basis for sexual identity, the mutagenic properties of X-irradiation and the isolation of the first tumour suppressor mutations. Drosophila researchers have developed an impressive array of sophisticated genetic techniques for analysis of gene function and genetic interactions. This review focuses on how these techniques can be utilised to study spermatogenesis in an organism with a generation time of 9 days and the capacity to introduce multiple mutant alleles into an individual organism in a relatively short time frame.

scholarly journals PRC1 coordinates timing of sexual differentiation of female primordial germ cells

Nature ◽  
2013 ◽  
Vol 495 (7440) ◽  
pp. 236-240 ◽  
Author(s):  
Shihori Yokobayashi ◽  
Ching-Yeu Liang ◽  
Hubertus Kohler ◽  
Peter Nestorov ◽  
Zichuan Liu ◽  
...  
Keyword(s):  
Germ Cells ◽  
Sexual Differentiation ◽  
Primordial Germ Cells

scholarly journals Sexual Differentiation and Primordial Germ Cell Distribution in the Early Horse Fetus

Animals ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 2422
Author(s):  
Dragos Scarlet ◽  
Stephan Handschuh ◽  
Ursula Reichart ◽  
Giorgia Podico ◽  
Robyn E. Ellerbrock ◽  
...  
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Sexual Differentiation ◽  
Primordial Germ Cells ◽  
Primordial Germ Cell ◽  
Tubular Structures ◽  
Micro Computed Tomography ◽  
Computed Tomography Scanning ◽  
Fetal Ovary ◽  
Well Differentiated

It was the aim of this study to characterize the development of the gonads and genital ducts in the equine fetus around the time of sexual differentiation. This included the identification and localization of the primordial germ cell population. Equine fetuses between 45 and 60 days of gestation were evaluated using a combination of micro-computed tomography scanning, immunohistochemistry, and multiplex immunofluorescence. Fetal gonads increased in size 23-fold from 45 to 60 days of gestation, and an even greater increase was observed in the metanephros volume. Signs of mesonephros atrophy were detected during this time. Tubular structures of the fetal testes were present from day 50 onwards, whereas cell clusters dominated in the fetal ovary. The genital ducts were well-differentiated and presented a lumen in all samples. No sign of mesonephric or paramesonephric duct degeneration was detected. Expression of AMH was strong in the fetal testes but absent in ovaries. Irrespective of sex, primordial germ cells selectively expressed LIN28. Migration of primordial germ cells from the mesonephros to the gonad was detected at 45 days, but not at 60 days of development. Their number and distribution within the gonad were influenced (p < 0.05) by fetal sex. Most primordial germ cells (86.8 ± 3.2% in females and 84.6 ± 4.7% in males) were characterized as pluripotent according to co-localization with CD117. However, only a very small percentage of primordial germ cells were proliferating (7.5 ± 1.7% in females and 3.2 ± 1.2% in males) based on co-localization with Ki67. It can be concluded that gonadal sexual differentiation in the horse occurs asynchronously with regard to sex but already before 45 days of gestation.

Germ-cell line and sexual differentiation in birds

1970 ◽  
Vol 259 (828) ◽  
pp. 73-90 ◽  
Keyword(s):  
Germ Cells ◽  
Sexual Differentiation ◽  
Germ Line ◽  
Primordial Germ Cells ◽  
Kinetic Studies ◽  
Blood Stream ◽  
Excretory Function ◽  
Germ Cell Line ◽  
Cytoplasmic Proteins ◽  
Secondary Formation

The present paper deals with recent investigations on the germ line and the sexual organogenesis in birds. The analysis has been limited to the following problems: origin of the germ line, physiology of the germinal epithelia, determinism of the migration of gonocytes, and differentiation of the germ cells during sexual organogenesis. It has been clearly established that in birds the germ line is precociously determined and that the anterior germinal crescent is a secondary formation. The hypothesis according to which the primary localization of the germ cells in birds (and more generally in Amniotes) would be posterior, is discussed. It appears to be the most plausible. The germinal epithelia are secretory and excretory organs. The excretory function is of the merocrine type. Kinetic studies of the protein turnover in the germinal epithelia demonstrate the existence of at least two categories of cytoplasmic proteins: structural proteins, and exportable proteins; the latter are excreted via protrusions of the epithelial cells. The hypothesis according to which the excretory function is associated with the attractive power exerted by the sexual primordia upon the migrating primordial germ cells is examined. The mechanism controlling the entry into the genital ridge of the gonocytes circulating in the embryonic blood stream is of a chemotactic nature. Finally, during early sexual organogenesis, as well as during later stages of sexual differentiation, the germ cells undergo important ultrastructural, histochemical and physiological (migratory properties) changes.

Early gonadal development and sexual differentiation in muskellunge (Esox masquinongy)

1997 ◽  
Vol 75 (8) ◽  
pp. 1262-1269 ◽  
Author(s):  
Feng Lin ◽  
Konrad Dabrowski ◽  
Lucy P. M. Timmermans
Keyword(s):  
Cross Section ◽  
Germ Cells ◽  
Sexual Differentiation ◽  
Resting State ◽  
Early Stage ◽  
Primordial Germ Cells ◽  
Gonadal Development ◽  
Mitotic Division ◽  
Early Prophase ◽  
Esox Masquinongy

Primordial germ cells (PGCs) were first identified in muskellunge (Esox masquinongy) of 14 mm total length (TL) 3 weeks post fertilization. At 32 mm TL, gonad strings were complete and formed a typical gonad shape in cross section. Blood vessels were first found in the gonads with Crossmon staining at 46 mm TL. Some of the PGCs underwent mitotic division at this stage. The ovarian sac started to develop in a fish of 82 mm TL, while the germ cells were still considered to be undifferentiated. In a fish of 138 mm TL, female gonads could be clearly identified from the ovarian sac and groups of oogonia, whereas in another type of gonad, the morphology of undifferentiated gonads was maintained. Germ cells became numerous in both sexes at 211 mm TL. Female gonads contained lobes with germ cells, including oogonia, early-prophase oocytes, and large oocytes. Spermatogonia and cells undergoing mitosis were observed in the testis. Ovaries in a fish of 250 mm TL were at the early stage of perinucleolus (early diplotene). Our observations indicate that in muskellunge (i) the PGCs remained in a resting state for up to 8 weeks post fertilization, (ii) gametogenesis occurred earlier in females than in males, (iii) the gonads developed from an undifferentiated stage directly into an ovary or testis, and (iv) the somatic elements in the gonads differentiated prior to the germ cells.

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