Conservation of chicken male germline by orthotopic transplantation of primordial germ cells from genetically distant donors†

2019 ◽  
Vol 101 (1) ◽  
pp. 200-207 ◽  
Author(s):  
Jitka Mucksová ◽  
Markéta Reinišová ◽  
Jiří Kalina ◽  
Barbora Lejčková ◽  
Jiří Hejnar ◽  
...  
Keyword(s):  
Genetic Variability ◽  
Genome Editing ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
Avian Species ◽  
Domestic Chicken ◽  
White Leghorn ◽  
Orthotopic Transplantation ◽  
Viable Offspring ◽  
Male Germline

Abstract Successful derivation and cultivation of primordial germ cells (PGCs) opened the way to efficient transgenesis and genome editing in the chicken. Furthermore, implantation of male PGCs from non-chicken galliform species into the chicken embryos resulted in cross-species germline chimeras and viable offspring. We have recently improved the PGC technology by demonstrating that chicken male PGCs transplanted into the testes of adult cockerel recipients mature into functional sperms. However, the availability of this orthotopic transplantation for cross-species transfer remains to be explored. Here we tested the capacity of genetically distant male PGCs to mature in the microenvironment of adult testes. We derived PGCs from the Chinese black-bone Silkie and transplanted them into infertile White Leghorn cockerels. Within 15–18 weeks after transplantation, we observed restoration of spermatogenesis in recipient cockerels and production of healthy progeny derived from the transplanted PGCs. Our findings also indicate the possibility of cross-species orthotopic transplantation of PGCs. Thus, our results might contribute to the preservation of endangered avian species and maintaining the genetic variability of the domestic chicken.

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.

Size-dependent isolation of primordial germ cells from avian species

2017 ◽  
Vol 84 (6) ◽  
pp. 508-516 ◽  
Author(s):  
Kyung M. Jung ◽  
Young M. Kim ◽  
Tamao Ono ◽  
Jae Y. Han
Keyword(s):  
Germ Cells ◽  
Primordial Germ Cells ◽  
Avian Species ◽  
Size Dependent

scholarly journals Investigation of the Guinea fowl and domestic fowl hybrids as potential surrogate hosts for avian cryopreservation programmes

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Mariann Molnár ◽  
Bence Lázár ◽  
Nikoletta Sztán ◽  
Barbara Végi ◽  
Árpád Drobnyák ◽  
...  
Keyword(s):  
Germ Cells ◽  
Domestic Fowl ◽  
Vascular System ◽  
Primordial Germ Cells ◽  
Research Work ◽  
Guinea Fowl ◽  
Hatching Rate ◽  
Migration Route ◽  
Sterile Hybrid ◽  
Viable Offspring

Abstract In the last decade, avian gene preservation research has focused on the use of the early precursors of the reproductive cells, the primordial germ cells (PGCs). This is because avian PGCs have a unique migration route through the vascular system which offers easy accessibility. Furthermore, culturing of the cells in vitro, freezing/thawing, reintegration into a recipient embryo and the development of the germ cells can be carried out in well-defined laboratory circumstances. The efficient recovery of the donor genotype and the frequency of germline transmission from the surrogate host animals are still areas which need further development. Thus, the aim of the present study was to investigate an infertile interspecific hybrid (recipient) as an appropriate host for primordial germ cells from native poultry breeds. Guinea fowl × chicken hybrids were produced, the crossing was repeated inversely. The phenotype, the hatching time, the hatching rate, the sex ratio, the presence of own germ cells, the fertility and the phenotype of viable hybrids and the incidence of chromosomal abnormalities of dead hybrid embryos were described. 6.65% viable offspring was obtained with crossing of Guinea fowl females with domestic fowl males. Crossing of domestic fowl hens with Guinea fowl male resulted in lower fertility, 0.14% viable offspring. Based on the investigations, the observed offspring from the successful crossing were sterile male hybrids, thus an extreme form of Haldane’s rule was manifested. The sterile hybrid male embryos were tested by injecting fluorescently labeled chicken PGCs. The integration rate of labeled PGCs was measured in 7.5-day, 14.5-day and 18.5-day old embryonic gonads. 50%, 5.3% and 2.4% of the injected hybrid embryos survived and 40%, 5.3% and 2.4% of the examined gonads contained fluorescent labeled donor PGCs. Therefore, these sterile hybrid males may be suitable recipients for male PGCs and possibly for female PGCs although with lower efficiency. This research work shows that the sterility of hybrids can be used in gene conservation to be a universal host for PGCs of different avian species.

scholarly journals Cryopreservation of Primordial Germ Cells (PGCs) from White Leghorn Embryos Using Commercial Cryoprotectants

2007 ◽  
Vol 44 (1) ◽  
pp. 73-77 ◽  
Author(s):  
Argono R. Setioko ◽  
Takahiro Tagami ◽  
Hideki Tase ◽  
Yoshiaki Nakamura ◽  
Kumiko Takeda ◽  
...  
Keyword(s):  
Germ Cells ◽  
Primordial Germ Cells ◽  
White Leghorn

Production of germ line chimera by transfer of primordial germ cells in the domestic chicken ()

1993 ◽  
Vol 40 (3) ◽  
pp. 509-519 ◽  
Author(s):  
A. Tajima ◽  
M. Naito ◽  
Y. Yasuda ◽  
T. Kuwana
Keyword(s):  
Germ Cells ◽  
Germ Line ◽  
Primordial Germ Cells ◽  
Domestic Chicken

scholarly journals Precise Genome Editing in Poultry and Its Application to Industries

Genes ◽  
2020 ◽  
Vol 11 (10) ◽  
pp. 1182
Author(s):  
Jin Se Park ◽  
Kyung Youn Lee ◽  
Jae Yong Han
Keyword(s):  
Genome Editing ◽  
Germ Cells ◽  
Food Industry ◽  
Recent Progress ◽  
Viral Vector ◽  
Primordial Germ Cells ◽  
Basic Sciences ◽  
Vector Systems ◽  
Novel Traits ◽  
Biomedical Industry

Poultry such as chickens are valuable model animals not only in the food industry, but also in developmental biology and biomedicine. Recently, precise genome-editing technologies mediated by the CRISPR/Cas9 system have developed rapidly, enabling the production of genome-edited poultry models with novel traits that are applicable to basic sciences, agriculture, and biomedical industry. In particular, these techniques have been combined with cultured primordial germ cells (PGCs) and viral vector systems to generate a valuable genome-edited avian model for a variety of purposes. Here, we summarize recent progress in CRISPR/Cas9-based genome-editing technology and its applications to avian species. In addition, we describe further applications of genome-edited poultry in various industries.

scholarly journals Single-Cell RNA Sequencing Revealed the Heterogeneity of Gonadal Primordial Germ Cells in Zebra Finch (Taeniopygia guttata)

2021 ◽  
Vol 9 ◽  
Author(s):  
Kyung Min Jung ◽  
Minseok Seo ◽  
Young Min Kim ◽  
Jin Lee Kim ◽  
Jae Yong Han
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
Zebra Finch ◽  
Germ Cells ◽  
Expression Patterns ◽  
Primordial Germ Cells ◽  
Cell Types ◽  
Avian Species ◽  
Single Population ◽  
Single Cell Rna Sequencing

Primordial germ cells (PGCs) are undifferentiated gametes with heterogeneity, an evolutionarily conserved characteristic across various organisms. Although dynamic selection at the level of early germ cell populations is an important biological feature linked to fertility, the heterogeneity of PGCs in avian species has not been characterized. In this study, we sought to evaluate PGC heterogeneity in zebra finch using a single-cell RNA sequencing (scRNA-seq) approach. Using scRNA-seq of embryonic gonadal cells from male and female zebra finches at Hamburger and Hamilton (HH) stage 28, we annotated nine cell types from 20 cell clusters. We found that PGCs previously considered a single population can be separated into three subtypes showing differences in apoptosis, proliferation, and other biological processes. The three PGC subtypes were specifically enriched for genes showing expression patterns related to germness or pluripotency, suggesting functional differences in PGCs according to the three subtypes. Additionally, we discovered a novel biomarker, SMC1B, for gonadal PGCs in zebra finch. The results provide the first evidence of substantial heterogeneity in PGCs previously considered a single population in birds. This discovery expands our understanding of PGCs to avian species, and provides a basis for further research.

scholarly journals Recent Advances and Future Perspectives of In Vivo Targeted Delivery of Genome-Editing Reagents to Germ cells, Embryos, and Fetuses in Mice

Cells ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 799 ◽  
Author(s):  
Masahiro Sato ◽  
Shuji Takabayashi ◽  
Eri Akasaka ◽  
Shingo Nakamura
Keyword(s):  
Gene Delivery ◽  
Genome Editing ◽  
Germ Cells ◽  
Targeted Delivery ◽  
Genetic Manipulation ◽  
Ex Vivo ◽  
Primordial Germ Cells ◽  
Nucleic Acid Delivery ◽  
Advantages And Disadvantages

The recently discovered clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) systems that occur in nature as microbial adaptive immune systems are considered an important tool in assessing the function of genes of interest in various biological systems. Thus, development of efficient and simple methods to produce genome-edited (GE) animals would accelerate research in this field. The CRISPR/Cas9 system was initially employed in early embryos, utilizing classical gene delivery methods such as microinjection or electroporation, which required ex vivo handling of zygotes before transfer to recipients. Recently, novel in vivo methods such as genome editing via oviductal nucleic acid delivery (GONAD), improved GONAD (i-GONAD), or transplacental gene delivery for acquiring genome-edited fetuses (TPGD-GEF), which facilitate easy embryo manipulation, have been established. Studies utilizing these techniques employed pregnant female mice for direct introduction of the genome-editing components into the oviduct or were dependent on delivery via tail-vein injection. In mice, embryogenesis occurs within the oviducts and the uterus, which often hampers the genetic manipulation of embryos, especially those at early postimplantation stages (days 6 to 8), owing to a thick surrounding layer of tissue called decidua. In this review, we have surveyed the recent achievements in the production of GE mice and have outlined the advantages and disadvantages of the process. We have also referred to the past achievements in gene delivery to early postimplantation stage embryos and germ cells such as primordial germ cells and spermatogonial stem cells, which will benefit relevant research.

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