Efficient system for preservation and regeneration of genetic resources in chicken: concurrent storage of primordial germ cells and live animals from early embryos of a rare indigenous fowl (Gifujidori)

2010 ◽  
Vol 22 (8) ◽  
pp. 1237 ◽  
Author(s):  
Yoshiaki Nakamura ◽  
Fumitake Usui ◽  
Daichi Miyahara ◽  
Takafumi Mori ◽  
Tamao Ono ◽  
...  
Keyword(s):  
Genetic Resources ◽  
Liquid Nitrogen ◽  
Early Development ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
White Leghorn ◽  
Same Sex ◽  
Efficient System ◽  
Male And Female ◽  
Early Embryos

The unique accessibility of chicken primordial germ cells (PGCs) during early development provides the opportunity to combine the reproduction of live animals with genetic conservation. Male and female Gifujidori fowl (GJ) PGCs were collected from the blood of early embryos, and cryopreserved in liquid nitrogen for >6 months until transfer. Manipulated GJ embryos were cultured until hatching; fertility tests indicated that they had normal reproductive abilities. Embryos from two lines of White Leghorn (24HS, ST) were used as recipients for chimera production following blood removal. The concentration of PGCs in the early embryonic blood of 24HS was significantly higher than in ST (P < 0.05). Frozen–thawed GJ PGCs were microinjected into the bloodstream of same-sex recipients. Offspring originating from GJ PGCs in ST recipients were obtained with a higher efficiency than those originating from GJ PGCs in 24HS recipients (23.3% v. 3.1%). Additionally, GJ progeny were successfully regenerated by crossing germline chimeras of the ST group. In conclusion, the cryogenic preservation of PGCs from early chicken embryos was combined with the conservation of live animals.

scholarly journals Protein expression reveals a molecular sexual identity of avian primordial germ cells at pre-gonadal stages

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Laura Soler ◽  
Sabine Alves ◽  
Aurélien Brionne ◽  
Aurore Jacques ◽  
Vanessa Guérin ◽  
...  
Keyword(s):  
Genetic Resources ◽  
Sexual Identity ◽  
Cell Fate ◽  
Germ Cells ◽  
Sex Chromosome ◽  
Sex Differentiation ◽  
Primordial Germ Cells ◽  
Specific Cell ◽  
Male And Female

AbstractIn poultry, in vitro propagated primordial germ cells (PGCs) represent an important tool for the cryopreservation of avian genetic resources. However, several studies have highlighted sexual differences exhibited by PGCs during in vitro propagation, which may compromise their reproductive capacities. To understand this phenomenon, we compared the proteome of pregonadal migratory male (ZZ) and female (ZW) chicken PGCs propagated in vitro by quantitative proteomic analysis using a GeLC-MS/MS strategy. Many proteins were found to be differentially abundant in chicken male and female PGCs indicating their early sexual identity. Many of the proteins more highly expressed in male PGCs were encoded by genes localised to the Z sex chromosome. This suggests that the known lack of dosage compensation of the transcription of Z-linked genes between sexes persists at the protein level in PGCs, and that this may be a key factor of their autonomous sex differentiation. We also found that globally, protein differences do not closely correlate with transcript differences indicating a selective translational mechanism in PGCs. Male and female PGC expressed protein sets were associated with differential biological processes and contained proteins known to be biologically relevant for male and female germ cell development, respectively. We also discovered that female PGCs have a higher capacity to uptake proteins from the cell culture medium than male PGCs. This study presents the first evidence of an early predetermined sex specific cell fate of chicken PGCs and their sexual molecular specificities which will enable the development of more precise sex-specific in vitro culture conditions for the preservation of avian genetic resources.

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.

72 PRESERVATION OF PORCINE GONOCYTES AT 4°C AND IN LIQUID NITROGEN - A PRESERVATION MODEL OF GENETIC RESOURCES IN DOMESTIC ANIMALS AND IN ENDANGERED SPECIES

2008 ◽  
Vol 20 (1) ◽  
pp. 117
Author(s):  
M. Fujihara ◽  
S. Goel ◽  
Y. Kimura ◽  
N. Minami ◽  
M. Yamada ◽  
...  
Keyword(s):  
Stem Cells ◽  
Endangered Species ◽  
Cell Viability ◽  
Genetic Resources ◽  
Liquid Nitrogen ◽  
Germ Cells ◽  
Domestic Animals ◽  
Spermatogonial Stem Cells ◽  
Freezing And Thawing ◽  
Neonatal Testis

Gonocytes are primitive germ cells that reside in neonatal testis and are believed to be progenitor-type stem cells that differentiate into spermatogonial stem cells. Because of their self-renewal ability, gonocytes may be one of the targets for cryopreservation of genetic resources in domestic animals and in endangered species. However, there are only a few reports regarding the preservation of gonocytes and spermatogonial stem cells isolated from the testis. In this experiment, porcine gonocytes were used as a model for preservation of genetic resources. Porcine testes were collected at 2–6 days after birth. They were divided into the 5 experimental groups for storage: (1) DMEM/F12 medium, (2) DMEM/F12 + 15 mm HEPES, (3) PBS, (4) PBS + 15 mm HEPES, and (5) Liquid-Free, and stored at 4�C for 24 h. The testes were minced by scissors and digested with 2-step enzyme treatments. The gonocytes were isolated by Percoll density gradients and recovered from the fraction between 50 and 60%. The viability of cells was assessed using trypan blue dye exclusion. To determine optimum cryopreservation conditions for gonocytes, 10% DMSO, 10% glycerol, and 0.07 mm sucrose were used as cryoprotectants. The isolated gonocytes were suspended in DMEM/F12 + 10% FBS containing cryoprotectant at 4�C, kept at –80�C overnight, and finally immersed in liquid nitrogen. After freezing and thawing of gonocytes, cells were examined for viability and then cultured in DMEM/F12 + 10% FBS in 5% CO2, 95% air at 37�C in humidified atmosphere. Identification of gonocytes was performed using a specific marker of gonocytes, a lectin Dolichos biflorus agglutinin (DBA; Goel et al. 2007 Biol. Reprod. 77, 127–137). The gonocytes were recovered from testes at the purity level of around 70%. Cell viability in average after storage of testes at 4�C was significantly higher in DMEM/F12 + HEPES (95.3%) and PBS + HEPES (89.8%) than in DMEM/F12 (73.9%), PBS (79.7%), and Liquid-Free (72.2%) (P < 0.05; ANOVA). The addition of HEPES in storage medium seemed to be effective for improving cell viability. The use of 10% DMSO and 0.07 mm sucrose as cryoprotectants supported high cell viability (74.4%) of gonocytes after freezing and thawing. The addition of glycerol had an adverse effect on cell viability after freezing (18.3%). When cells were cultured, gonocytes started to form colonies after 3 days and continued to proliferate for at least 7 days in culture. These colonies showed DBA affinity and maintained their nature as gonocytes. The viability of gonocytes can be maintained in the testis at 4�C for at least 24 h and after freezing and thawing. The stored gonocytes successfully proliferated in culture for at least 7 days. In conclusion, these results may provide useful information for short-term storage of primitive germ cells and preservation of genetic resources in domestic animals and in endangered species. It may also have implications for assisted reproductive technology in humans.

Early development of primordial germ cells in Pacific bluefin tuna Thunnus orientalis

2019 ◽  
Vol 131 ◽  
pp. 106-112 ◽  
Author(s):  
Kentaro Higuchi ◽  
Rie Goto ◽  
Junpei Konishi ◽  
Yoshiaki Ina ◽  
Yukinori Kazeto ◽  
...  
Keyword(s):  
Early Development ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
Bluefin Tuna ◽  
Pacific Bluefin Tuna ◽  
Thunnus Orientalis

scholarly journals Localization of primordial germ cells or their precursors in stage X blastoderm of chickens and their ability to differentiate into functional gametes in opposite-sex recipient gonads

Reproduction ◽  
2001 ◽  
pp. 547-552 ◽  
Author(s):  
M Naito ◽  
A Sano ◽  
Y Matsubara ◽  
T Harumi ◽  
T Tagami ◽  
...  
Keyword(s):  
Germ Cells ◽  
Marginal Zone ◽  
Primordial Germ Cells ◽  
Donor Cell ◽  
Same Sex ◽  
White Leghorn Chicken ◽  
Central Disc ◽  
Opposite Sex ◽  
A Cell ◽  
Blastodermal Cells

This study was performed to determine the distribution of primordial germ cells and their precursors in stage X blastoderm of chickens. The blastoderm (Barred Plymouth Rock chickens) isolated from the yolk was separated into three portions: the central disc, the marginal zone and the area opaca. The dissociated blastodermal cells derived from the central disc, marginal zone and area opaca were transferred into a recipient blastoderm (White Leghorn chicken) from which a cell cluster was removed from the centre of the central disc. The manipulated embryos were cultured in host eggshells until hatching. The chicks were raised until sexual maturity and test mated with Barred Plymouth Rock chickens to assess the donor cell contribution to the recipient germline. Germline chimaeric chickens were produced efficiently (46.7%, 7/15) when the blastodermal cells derived from the central disc were transferred into recipient embryos of the same sex, whereas no germline chimaeric chickens were produced when the blastodermal cells derived from the marginal zone or area opaca were transferred into recipient embryos of the same sex (0/12). Germline chimaeric chickens were also produced by transfer of blastodermal cells derived from the central disc (6.7%, 1/15), marginal zone (10.0%, 1/10) or area opaca (11.1%, 1/9) into recipient embryos of the opposite sex. It is concluded that primordial germ cells are induced during or shortly after stage X and that the cells derived from the central disc have the highest potential to give rise to germ cells. Cells derived from the marginal zone and area opaca can also give rise to germ cells, although the frequency is low.

scholarly journals Fine-tuning evolution: germ-line epigenetics and inheritance

Reproduction ◽  
2013 ◽  
Vol 146 (1) ◽  
pp. R37-R48 ◽  
Author(s):  
Jessica M Stringer ◽  
Sanna Barrand ◽  
Patrick Western
Keyword(s):  
Germ Cells ◽  
Cell Function ◽  
Germ Line ◽  
Primordial Germ Cells ◽  
Epigenetic Reprogramming ◽  
Fine Tuning ◽  
Male And Female ◽  
Epigenetic Information

In mice, epiblast cells found both the germ-line and somatic lineages in the developing embryo. These epiblast cells carry epigenetic information from both parents that is required for development and cell function in the fetus and during post-natal life. However, germ cells must establish an epigenetic program that supports totipotency and the configuration of parent-specific epigenetic states in the gametes. To achieve this, the epigenetic information inherited by the primordial germ cells at specification is erased and new epigenetic states are established during development of the male and female germ-lines. Errors in this process can lead to transmission of epimutations through the germ-line, which have the potential to affect development and disease in the parent's progeny. This review discusses epigenetic reprogramming in the germ-line and the transmission of epigenetic information to the following generation.

scholarly journals Production of Germline Chimeric Quail by Transfer of Gonadal Primordial Germ Cells Preserved in Liquid Nitrogen.

1998 ◽  
Vol 35 (6) ◽  
pp. 321-328 ◽  
Author(s):  
Il-Kuk CHANGI ◽  
Mitsuru NAITO ◽  
Takashi KUWANA ◽  
Makoto MIZUTANI ◽  
Michiharu SAKURAI
Keyword(s):  
Liquid Nitrogen ◽  
Germ Cells ◽  
Primordial Germ Cells

scholarly journals Restoration of Genetic Resources from Ehime Native Chicken via Transferred Primordial Germ Cells (PGCs)

2001 ◽  
Vol 38 (4) ◽  
pp. 302-307 ◽  
Author(s):  
Hiroki Furuta ◽  
Keiji Kinoshita ◽  
Yoshizane Maeda ◽  
Noboru Fujihara
Keyword(s):  
Genetic Resources ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
Native Chicken

scholarly journals Knockdown of DEAD-box helicase 4 (DDX4) decreases the number of germ cells in male and female chicken embryonic gonads

2019 ◽  
Vol 31 (5) ◽  
pp. 847
Author(s):  
Nana Aduma ◽  
Hiroe Izumi ◽  
Shusei Mizushima ◽  
Asato Kuroiwa
Keyword(s):  
Germ Cells ◽  
Primordial Germ Cells ◽  
Retroviral Vectors ◽  
Ovary Development ◽  
Aromatase Expression ◽  
Male And Female ◽  
Chicken Embryos ◽  
Dead Box ◽  
Related Transcription Factor ◽  
Cytochrome P450 Family

DEAD-box helicase 4 (DDX4; also known as vasa) is essential for the proper formation and maintenance of germ cells. Although DDX4 is conserved in a variety of vertebrates and invertebrates, its roles differ between species. This study investigated the function of DDX4 in chicken embryos by knocking down its expression using retroviral vectors that encoded DDX4-targeting microRNAs. DDX4 was effectively depleted invitro and invivo via this approach. Male and female gonads of DDX4-knockdown embryos contained a decreased number of primordial germ cells, indicating that DDX4 is essential to maintain a normal level of these cells in chicken embryos of both sexes. Expression of doublesex and mab-3 related transcription factor 1 (DMRT1) and sex determining region Y-box 9 (SOX9), which are involved in testis determination and differentiation, was normal in male gonads of DDX4-knockdown embryos. In contrast, expression of cytochrome P450 family 19 subfamily A member 1 (CYP19A1), which encodes aromatase and is essential for ovary development, was significantly decreased in female gonads of DDX4-knockdown embryos. Expression of forkhead box L2 (FOXL2), which plays an important role in ovary differentiation, was also slightly reduced in DDX4-knockdown embryos, but not significantly. Based on several pieces of evidence FOXL2 was hypothesised to regulate aromatase expression. The results of this study indicate that aromatase expression is also regulated by several additional pathways.

Embryological Development of Primordial Germ-cells in the Mouse: Influence of a New Mutation, Wj1

Development ◽  
1957 ◽  
Vol 5 (4) ◽  
pp. 396-403
Author(s):  
Beatrice Mintz
Keyword(s):  
Germ Cells ◽  
Present Report ◽  
Primordial Germ Cells ◽  
Embryological Development ◽  
New Mutation ◽  
Preliminary Note ◽  
Male And Female ◽  
Embryonic Life ◽  
Mutant Genes ◽  
Pleiotropic Mutant

The pleiotropic mutant genes W and Wv are alleles of w in the mouse, and produce anaemia, absence of fur pigmentation, and sterility in homozygotes (review by Russell, 1954). Germ-cells of both male and female homozygotes are lacking or drastically reduced in numbers at birth, the genotypes being identifiable through the concurrent anaemia. The developmental basis for this sterility was therefore sought in embryonic life and has been described (Mintz & Russell, 1955, 1957). Recently, a new mutation, Wj, with comparable effects in the homozygote, arose at the same locus. Evidence that it is an allele of the W-series, but different from W or Wv, will be presented elsewhere (Russell, Lawson, & Schabtach, in preparation). In the present report, the early abnormalities characterizing WjWj will be traced and compared with those produced by the other mutant alleles, and will be considered in relation to the problems of germ-cell origin and pleiotropism. A preliminary note (Mintz, 1957) has appeared on the study.

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