scholarly journals Comparison of the Transcriptomic and Epigenetic Profiles of Gonadal Primordial Germ Cells of White Leghorn and Green-Legged Partridgelike Chicken Embryos

Genes ◽  
2021 ◽  
Vol 12 (7) ◽  
pp. 1090
Author(s):  
Aleksandra Dunislawska ◽  
Maria Siwek ◽  
Katarzyna Stadnicka ◽  
Marek Bednarczyk
Keyword(s):  
Embryonic Development ◽  
Germ Cells ◽  
Developmental Stages ◽  
Primordial Germ Cells ◽  
Genetic Material ◽  
Reproductive Traits ◽  
Germline Stem Cells ◽  
White Leghorn ◽  
Methylation Analysis ◽  
Global Methylation

The Green-legged Partridgelike fowl is a native, dual-purpose Polish chicken. The White Leghorn has been intensively selected for several decades to mainly improve reproductive traits. Primordial germ cells (PGCs) represent the germline stem cells in chickens and are the only cells that can transfer the information stored in the genetic material from generation to generation. The aim of the study was to carry out a transcriptomic and an epigenetic comparison of the White Leghorn and Green-legged Partridgelike gonadal PGCs (gPGCs) at three developmental stages: days 4.5, 8, and 12 of the embryonic development. RNA and DNA were isolated from collected gPGCs. The RNA was further subjected to microarray analysis. An epigenetic analysis was performed based on the global methylation analysis and qMSP method for the particular silenced genes demonstrated in transcriptomic analysis. Statistically significant differences between the gPGCs from both breeds were detected on the day 8 of embryonic development. Global methylation analysis showed significant changes at the methylation level in the White Leghorn gPGCs on day 8 of embryonic development. The results suggest faster development of Green-legged Partridgelike embryos as compared to White Leghorn embryos. Changes in the levels of gene expression during embryonic development are determined by genetic and environmental factors, and this variability is influenced by breed and gender.

Function of TGF-?2 in the growth of chicken primordial germ cells and germinal ridge stroma cells during embryonic development

2004 ◽  
Vol 301A (4) ◽  
pp. 290-296 ◽  
Author(s):  
Tsuyoshi Fujioka ◽  
Tomoki Soh ◽  
Noboru Fujihara ◽  
Masa-Aki Hattori
Keyword(s):  
Embryonic Development ◽  
Germ Cells ◽  
Primordial Germ Cells

scholarly journals NUP50 is necessary for the survival of primordial germ cells in mouse embryos

Reproduction ◽  
2016 ◽  
Vol 151 (1) ◽  
pp. 51-58 ◽  
Author(s):  
Eunsook Park ◽  
Bobae Lee ◽  
Bruce E Clurman ◽  
Keesook Lee
Keyword(s):  
Embryonic Development ◽  
Germ Cells ◽  
Nuclear Pore Complex ◽  
Essential Role ◽  
Primordial Germ Cells ◽  
Mouse Embryos ◽  
Nuclear Pore ◽  
Progressive Loss ◽  
And Function ◽  
Deficient Mice

Nucleoporin 50 kDa (NUP50), a component of the nuclear pore complex, is highly expressed in male germ cells, but its role in germ cells is largely unknown. In this study, we analyzed the expression and function of NUP50 during the embryonic development of germ cells using NUP50-deficient mice. NUP50 was expressed in germ cells of both sexes at embryonic day 15.5 (E15.5), E13.5, and E12.5. In addition, NUP50 expression was also detected in primordial germ cells (PGCs) migrating into the genital ridges at E9.5. The gonads of Nup50−/− embryos of both sexes contained few PGCs at both E11.5 and E12.5 and no developing germ cells at E15.5. The migratory PGCs in Nup50−/− embryos at E9.5 showed increased apoptosis but a normal rate of proliferation, resulting in the progressive loss of germ cells at later stages. Taken together, these results suggest that NUP50 plays an essential role in the survival of PGCs during embryonic development.

scholarly journals Determination of production capacity of circulated primordial germ cells (circulated-PGCs) of KUB chicken using lysis buffer ammonium chloride potassium (ACK)

2016 ◽  
Vol 21 (1) ◽  
pp. 55
Author(s):  
Soni Sopiyana ◽  
Iman Supriatna ◽  
M. Agus Setiadi ◽  
Mohamad Fahrudin
Keyword(s):  
Embryonic Development ◽  
Ammonium Chloride ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
Production Capacity ◽  
Development Stage ◽  
Simple Method ◽  
Average Production ◽  
Short Period ◽  
Lysis Buffer

<p class="abstrak2">In poultry embryos, primordial germ cells (PGCs) are progenitor cells for gametes, which have unique migration pathway. Primordial germ cells arise from epiblast in germinal crescent and circulate through the bloodstream for a short period of time, then leave blood vessel to migrate toward gonads. The aim of this study was to determine the potential production capacity of circulated-PGCs of KUB chicken at different developmental stages of embryo using a rapid and simple method. Seventy five KUB chicken fertile eggs were divided into five groups and incubated at 38.5 <sup>0</sup>C with a humidity of 60%. Hatching was set to the embryonic development stage of 14-18. The blood was collected through dorsal aorta using micropipette under microscope. The collected blood was placed in a 1.5 ml eppendorf tube which was previously filled with 100 µl phosphate buffered saline without Ca<sup>2+</sup> and Mg<sup>2+</sup> (PBS-) mixed with fetal bovine serum (FBS) with a ratio of 90%:10%. The PGCs were purified using lysis buffer ammonium chloride potassium method. The results showed that average production of circulated-PGCs per embryo of KUB chicken were significantly affected by stage of embryonic development (P &lt;0.05). The average production of circulated-PGCs at stage 14, 15, 16, 17, and 18 were 37.9; 53.5; 49.8; 38.3; and 33.5 respectively. The number of circulated-PGCs was not different among stages 14, 17 nor 18. The highest number of circulated-PGCs of KUB chicken was obtained at stage 15, so that the isolation and collection of PGCs through the blood circulation was recommended in stage 15.</p><strong>Key Words: </strong>KUB Chicken, PGCs, Embryonic Development Stage, Ammonium Chloride Potassium

scholarly journals When Regenerative Medicine Faces the Challenges of Reproductive Medicine: A Review Study on Recent Advances in the Strategies for Derivation of Gametes from Stem Cells

2020 ◽  
pp. 42-52
Author(s):  
María Gil Juliá ◽  
José V. Medrano
Keyword(s):  
Stem Cells ◽  
Germ Cells ◽  
Developmental Stages ◽  
Primordial Germ Cells ◽  
Embryonic Stem ◽  
Differentiation Strategy ◽  
Stem Cell Source ◽  
Potential Applications ◽  
Key Events

The murine model has allowed for the replication of all developmental stages of the mammalian germline in vitro, from embryonic stem cells to epiblast cells, primordial germ cells, and finally into functional haploid gametes. However, because of interspecies differences between mice and humans, these results are yet to be replicated in our species. Reports on the use of stem cells as a source of gametes, retrieved from public scientific databases, were analysed and classified according to the animal model used, the stem cell source and type, the differentiation strategy, and its potential application. This review offers a comprehensive compilation of recent publications of key events in the derivation of germ cells and gametogenesis in vitro, in both mice and human models. Additionally, studies intending to replicate the different stages in human cells in vitro, in order to obtain cells with a phenotype akin to functional human gametes, are also depicted. The authors present options for deriving gametes from stem cells in vitro and different reproductive options for specific groups of patients. Lastly, the potential applications of in vitro human gametogenesis are evaluated as well as the main limitations of the techniques employed. Even though it appears that we are far from being able to obtain gametes from pluripotent stem cells in vitro as a viable reproductive option, its current academic and clinical implications are extremely promising.

63 THE EFFECT OF A MODIFIED CRYOPRESERVATION METHOD ON THE VIABILITY OF FROZEN–THAWED PRIMORDIAL GERM CELL ON THE KOREAN NATIVE CHICKEN (Ogye)

2014 ◽  
Vol 26 (1) ◽  
pp. 145
Author(s):  
H. Kim ◽  
D. H. Kim ◽  
J. Y. Han ◽  
S. B. Choi ◽  
Y.-G. Ko ◽  
...  
Keyword(s):  
Germ Cells ◽  
Germ Line ◽  
New Technology ◽  
Primordial Germ Cells ◽  
Primordial Germ Cell ◽  
Genetic Material ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Method ◽  
Native Chicken ◽  
Korean Native Chicken

Cryopreservation of poultry semen has been reported, but preservation of female genetic material has not been possible because of the unique anatomical and physiological characteristics of the avian egg. Thus, conservation of genetic material in chickens was attempted by preserving primordial germ cells (PGC) in LN2. This study established a method for preserving chicken PGC that enables long-term storage in LN2 for preservation of species. The purpose of this study was to clarify the effects of fetal bovine serum (FBS) or chicken serum (CS) treatment on the viability of cryopreserved PGC in the Korean native chicken (Ogye). Primordial germ cells were separated from a germinal gonad using a fine glass micropipette under a microscope and were suspended in a freezing medium containing freezing and protecting agents [e.g. dimethyl sulfoxide (DMSO) and ethylene glycol (EG)]. The PGC were then purified using the magnetic-activated cell sorting (MACS) method. The viability of the PGC in both groups was determined by the trypan blue exclusion method. The values of the 0, 5, 10, and 15% DMSO plus FBS treatment were 21.6, 30.36, 36.42, 50.39, and 48.36%, respectively. The viability of PGC after freeze-thawing was significantly higher for the 10% EG plus FBS treatment than for the 10% EG plus CS treatment (P < 0.05; 64.36 v. 50.66%). This study established a method for preserving chicken PGC that enables systematic storage and labelling of cryopreserved PGC in LN2 at a germplasm repository and ease of entry into a database. In the future, the importance of this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germ line chimeras.

The Relation of the Interrenal Blastema to the Origin of the Somatic Tissues of the Gonad in Frog Tadpoles

Development ◽  
1954 ◽  
Vol 2 (4) ◽  
pp. 275-289
Author(s):  
Enrico Vannini ◽  
Armando Sabbadin
Keyword(s):  
Germ Cells ◽  
Developmental Stages ◽  
Primordial Germ Cells ◽  
Lateral Position ◽  
Medullary Tissue ◽  
Somatic Tissues ◽  
Frog Tadpoles ◽  
Dorsal Mesentery

As long ago as 1941 and 1942 one of us (Vannini) found in a series of developmental stages of frog tadpoles that the somatic components of the medullary tissue of the gonad have their origin in the interrenal blastema, and not, as was then generally supposed, in the mesonephric blastema. In the earliest stages examined at that time the gonad rudiment had the structure of ‘paired genital ridges’, lying at each side of the dorsal mesentery, and were furnished with primordial germ-cells, but were still without medullary tissue. The interrenal blastema occupied a median site in the tadpole's body, ventral to the aorta and dorsal to the two subcardinal veins. The mesonephric blastemata appeared distinctly separate from the interrenal rudiment, because they were situated in a more lateral position, contiguous with the Wolffian ducts. In later stages the medullary tissue (‘medulla’) penetrated within the genital ridges.

Lentiviral vector transfection of chicken embryo primordial germ cells in vitro

2008 ◽  
Vol 5 (1) ◽  
pp. 1-5
Author(s):  
Ding Hong-Mei ◽  
Xu Shi-Yong ◽  
Shao Gen-Bao ◽  
Sun Yan ◽  
Wang Meng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Gene Transfer ◽  
Germ Cells ◽  
Lentiviral Vector ◽  
Transfection Efficiency ◽  
Primordial Germ Cells ◽  
Embryonic Stem ◽  
Germline Stem Cells ◽  
Transfer Vectors

AbstractLentiviruses as gene transfer vectors have been used successfully to transfect mammal embryonic stem cells and germline stem cells, but this has not been attempted in avian primordial germ cells (PGCs). PGCs were isolated from the gonads of Isa-brown chicken embryos at stage 28 and co-cultured with gonadal stroma cells. A lentiviral vector pLenti-CMV-EGFP was constructed and the virus harvested by cotransfecting 293FT cells with the vector and packaging plasmids. Concentrated lentiviruses were used to transfect chicken PGCs, the transfection efficiency was up to 24.19%.

scholarly journals Primordial germ cells do not migrate along nerve fibres in marmoset monkey and mouse embryos

Reproduction ◽  
2019 ◽  
pp. 101-109 ◽  
Author(s):  
E Wolff ◽  
M M Suplicki ◽  
R Behr
Keyword(s):  
Germ Cells ◽  
Peripheral Nerves ◽  
Developmental Stages ◽  
Primordial Germ Cells ◽  
Spatial Association ◽  
General Mechanism ◽  
Human Embryos ◽  
General Strategy ◽  
Marmoset Monkey ◽  
Specific Finding

Primordial germ cells (PGCs) are the embryonic precursors of spermatozoa and eggs. In mammals, PGCs arise early in embryonic development and migrate from their tissue of specification over a significant distance to reach their destinations, the genital ridges. However, the exact mechanism of translocation is still debated. A study on human embryos demonstrated a very close spatial association between migrating PGCs and developing peripheral nerves. Thus, it was proposed that peripheral nerves act as guiding structures for migrating PGCs. The goal of the present study is to test whether the association between nerves and PGCs may be a human-specific finding or whether this represents a general strategy to guide PGCs in mammals. Therefore, we investigated embryos of different developmental stages from the mouse and a non-human primate, the marmoset monkey (Callithrix jacchus), covering the phase from PGC emergence to their arrival in the gonadal ridge. Embryo sections were immunohistochemically co-stained for tubulin beta-3 chain (TUBB3) to visualise neurons and Octamer-binding protein 4 (OCT4 (POU5F1)) as marker for PGCs. The distance between PGCs and the nearest detectable neuron was measured. We discovered that in all embryos analysed of both species, the majority of PGCs (>94%) was found at a minimum distance of 50 µm to the closest neuron and, more importantly, that the PGCs had reached the gonads before any TUBB3 signal could be detected in the vicinity of the gonads. In conclusion, our data indicate that PGC migration along peripheral nerves is not a general mechanism in mammals.

scholarly journals The reversible developmental unipotency of germ cells in chicken

Reproduction ◽  
2010 ◽  
Vol 139 (1) ◽  
pp. 113-119 ◽  
Author(s):  
Jin Gyoung Jung ◽  
Young Mok Lee ◽  
Jin Nam Kim ◽  
Tae Min Kim ◽  
Ji Hye Shin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Germ Cell ◽  
Germ Cells ◽  
Developmental Stages ◽  
Production Systems ◽  
Primordial Germ Cells ◽  
Transmission Efficiency ◽  
Germline Transmission ◽  
Adult Testis

We recently developed bimodal germline chimera production approaches by transfer of primordial germ cells (PGCs) or embryonic germ cells (EGCs) into embryos and by transplantation of spermatogonial stem cells (SSCs) or germline stem cells (GSCs) into adult testes. This study was undertaken to investigate the reversible developmental unipotency of chicken germ cells using our established germline chimera production systems. First, we transferred freshly isolated SSCs from adult testis or in vitro cultured GSCs into stage X and stage 14–16 embryos, and we found that these transferred SSCs/GSCs could migrate to the recipient embryonic gonads. Of the 527 embryos that received SSCs or GSCs, 135 yielded hatchlings. Of 17 sexually mature males (35.3%), six were confirmed as germline chimeras through testcross analysis resulting in an average germline transmission efficiency of 1.3%. Second, PGCs/EGCs, germ cells isolated from embryonic gonads were transplanted into adult testes. The EGC transplantation induced germline transmission, whereas the PGC transplantation did not. The germline transmission efficiency was 12.5 fold higher (16.3 vs 1.3%) in EGC transplantation into testis (EGCs to adult testis) than that in SSC/GSC transfer into embryos (testicular germ cells to embryo stage). In conclusion, chicken germ cells from different developmental stages can (de)differentiate into gametes even after the germ cell developmental clock is set back or ahead. Use of germ cell reversible unipotency might improve the efficiency of germ cell-mediated germline transmission.

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