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 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.

Simple method for isolation of primordial germ cells from chick embryos

1992 ◽  
Vol 16 (9) ◽  
pp. 853-857 ◽  
Author(s):  
I CHANG ◽  
A TAJIMA ◽  
Y YASUDA ◽  
T CHIKAMUNE ◽  
T OHNO
Keyword(s):  
Germ Cells ◽  
Primordial Germ Cells ◽  
Chick Embryos ◽  
Simple Method

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 Generation of transgenic quails by in vivo transfection of primordial germ cells

2019 ◽  
Author(s):  
Olivier Serralbo ◽  
Nadège Véron ◽  
Caitlin Cooper ◽  
Marie-Julie Dejardin ◽  
Timothy Doran ◽  
...  
Keyword(s):  
Germ Cells ◽  
Genome Engineering ◽  
Genetic Model ◽  
Technological Development ◽  
Primordial Germ Cells ◽  
Basic Research ◽  
Short Period ◽  
Wide Range ◽  
Infinite Variety

AbstractDuring development, dynamic changes in tissue shapes known as morphogenesis result from the exquisite orchestration of signalling pathways, cell–cell interactions, cell divisions, and coordinated movements. How cells within embryos adopt a vast array of cell fates and tissue shapes in such an ever-changing environment has fascinated scientists for generations, yet the ability to observe and characterize those rapid changes has proven technically challenging in higher vertebrates. The japanese quail (Coturnix coturnix japonica) is an attractive model where basic rules driving morphogenesis in amniotes can be deciphered using genetic approaches. Similar to the more popular chicken model, the quail embryo is easily accessible to a wide range of manipulations and live imaging. A decisive asset of quail over chicken is a much shorter life cycle, which makes its use as genetic model for basic research extremely appealing. To date, all existing transgenic quail lines were generated using replication-deficient lentiviruses, but diverse limitations of this approach have hindered the widespread expansion of such technology. Here, we successfully used a plasmid-based in vivo transfection of quail primordial germ cells (PGCs) to generate a number of transgenic quail lines over a short period of time. The plasmid-based approach is simple, efficient and it allows using the infinite variety of genome engineering approaches developed in other models, such as strategies to facilitate transgenic bird selection, shown here. This major technological development completes the vast panel of techniques applicable to the avian model, making it one of the most versatile experimental systems available.

Differentiation of primordial germ cells in the embryonic development of Thermobia domestica, Pack. (Thysanura): an ultrastructural study

Development ◽  
1977 ◽  
Vol 38 (1) ◽  
pp. 93-114
Author(s):  
Jerzy Klag
Keyword(s):  
Embryonic Development ◽  
Germ Cells ◽  
Morphological Changes ◽  
Sex Differentiation ◽  
Primordial Germ Cells ◽  
Postembryonic Development ◽  
Thermobia Domestica ◽  
Dense Bodies ◽  
Accessory Nuclei ◽  
High Degree

The primordial germ cells (PGCs) of Thermobia domestica undergo some morphological changes during the embryonic development. Most conspicuous are the changes in the ultrastructure of the nucleus, whose envelope shows a high degree of activity. Two types of vesicles bleb off from the nucleus; the ones with the light interior are called the accessory nuclei, the others, with electron-opaque contents, have been termed the dense bodies. The nucleolus, initially clustered at the nucleus centre, undergoes dispersion and assembles again towards the end of embryonic development. At the same time, the sex differentiation of PGCs takes place. It is preceded by an increase in the activity of Golgi complexes and in the volume of lysosomes and lamellar bodies, the latter giving rise to lipid droplets. At the early stages of postembryonic development, preoogonia and prespermatogonia can readily be distinguished. Preoogonia have a wavy-surfaced nucleus and their cytoplasm contains dense bodies. In prespermatogonia, the nucleus is spherical with smooth envelope and there are no dense bodies in the cytoplasm. Throughout the period studied there occur nucleolus-like bodies and nuage material considered to be the germ-cell determinants in this species.

Developmental windows and origins of the chemical cues mediating hatching responses to injured conspecific eggs in the common frog (Rana temporaria)

2008 ◽  
Vol 86 (6) ◽  
pp. 447-455 ◽  
Author(s):  
A.-L. Mandrillon ◽  
P. Saglio
Keyword(s):  
Embryonic Development ◽  
Critical Period ◽  
Chemical Cues ◽  
Rana Temporaria ◽  
Morphological Plasticity ◽  
Development Stage ◽  
Common Frog ◽  
Embryonic Exposure ◽  
Short Period ◽  
Frog Rana Temporaria

In amphibians, embryonic exposure to chemical cues resulting from a predation event on conspecific eggs can influence hatching traits. However, there is no information on the precise origin of the active substances, or on the critical period of embryonic development mediating such a process. In this context, common frog ( Rana temporaria L., 1758) eggs were exposed at Gosner stage 2, 16, or 20 to chemical cues simulating predation on whole eggs, jelly envelopes, or embryos. Embryonic movement rate, hatching time, and developmental stage at hatching appeared unaffected by the nature of the treatment. In contrast, the embryonic treatments strongly affected the morphology of hatchlings, with the groups exposed to crushed whole eggs and jelly envelopes showing longer (exposures at stages 16 and 20) and deeper (exposure at stage 20) tails than their unexposed counterparts. In addition, exposure at stage 20 to crushed embryos also produced hatchlings with longer tails than the controls. Thus, morphological plasticity at hatching can result from a relatively short period of embryonic exposure to conspecific chemical cues. This critical period occurs at the completion of neurulation (stage 16), with the most marked effects resulting from an exposure at the last stage of embryonic development (stage 20).

scholarly journals Effects of W (c-kit) gene mutation on gametogenesis in male mice: agametic tubular segments in Wf/Wf testes

Development ◽  
1992 ◽  
Vol 114 (4) ◽  
pp. 861-867 ◽  
Author(s):  
U. Koshimizu ◽  
D. Watanabe ◽  
Y. Tajima ◽  
Y. Nishimune
Keyword(s):  
Stem Cell ◽  
Embryonic Development ◽  
Germ Cells ◽  
Seminiferous Tubule ◽  
Primordial Germ Cells ◽  
Mutant Mice ◽  
Tyrosine Kinase Receptor ◽  
Male Mice ◽  
Kit Gene ◽  
Development And Differentiation

Mutations of the W (c-kit) gene, which encodes a transmembrane tyrosine kinase receptor, affect the development and differentiation of many types of stem cell. Most homozygous W mutant mice are sterile, due to a lack of germ cells arising during embryonic development, but one of the notable exceptions is Wf/Wf mice, which are fully fertile in both sexes. In order to elucidate the effects of the Wf mutation on spermatogenesis, postnatal spermatogenesis in Wf/Wf mice was histologically examined. The number of gonocytes at birth was significantly reduced and small portions of agametic seminiferous tubule segments were observed in mutant mice. It is suggested that this is due to a deficiency of primordial germ cells (PGC). Other than the agametic tubules, there was no evidence of reduced spermatogenesis after birth. These results indicate that the function of the W (c-kit) gene is more necessary for the development of PGC than for postnatal germ cells.

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