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

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.

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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)

Reproduction Fertility and Development ◽

10.1071/rd10056 ◽

2010 ◽

Vol 22 (8) ◽

pp. 1237 ◽

Cited By ~ 26

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.

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

Biology of Reproduction ◽

10.1093/biolre/ioz064 ◽

2019 ◽

Vol 101 (1) ◽

pp. 200-207 ◽

Cited By ~ 3

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.

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Hexachlorobenzene toxicity in the rat ovary: II. Ultrastructure induced by medium (10 mg/kg) dose exposure

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010012374x ◽

1992 ◽

Vol 50 (1) ◽

pp. 668-669

Author(s):

Amreek Singh ◽

Warren G. Foster ◽

Anna Dykeman ◽

David C. Villeneuve

Keyword(s):

Germ Cells ◽

Primordial Germ Cells ◽

Surface Epithelium ◽

Morphological Parameters ◽

Comprehensive Investigation ◽

Ovary Surface Epithelium ◽

Rat Ovary ◽

High Doses

Hexachlorobenzene (HCB) is a known toxicant that is found in the environment as a by-product during manufacture of certain pesticides. This chlorinated chemical has been isolated from many tissues including ovary. When administered in high doses, HCB causes degeneration of primordial germ cells and ovary surface epithelium in sub-human primates. A purpose of this experiment was to determine a no-effect dose of the chemical on the rat ovary. The study is part of a comprehensive investigation on the effects of the compound on the biochemical, hematological, and morphological parameters in the monkey and rat.

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