Size-dependent isolation of primordial germ cells from avian species

Jung, Kyung Min; Kim, Young Min; Ono, Tamao; Han, Jae Yong

doi:10.1002/mrd.22802

Cited by 15 publications

(12 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Additionally, DAZL::GFP germ cells at E2.5, E6, E8, and 1 week post-hatch were collected to observe the expression patterns of DNA methylation-related and mitosis-related genes from blood-circulating PGCs up to 1 week after hatching. Whole blood cells were isolated from the dorsal aorta of each embryo using a glass micropipette under a microscope from 70 to 80 embryos at E2.5 [ 36 ]. The gonads or testes were microscopically dissected and pooled from 35 embryos, 11 embryos, and 3 chicks at E6, E8, and 1 week post-hatch, respectively (Additional file 1 : Table S1).…”

Section: Methodsmentioning

confidence: 99%

Single-cell RNA sequencing of mitotic-arrested prospermatogonia with DAZL::GFP chickens and revealing unique epigenetic reprogramming of chickens

Choi

Jung

Rengaraj

et al. 2022

J Animal Sci Biotechnol

Self Cite

View full text Add to dashboard Cite

Background Germ cell mitotic arrest is conserved in many vertebrates, including birds, although the time of entry or exit into quiescence phase differs. Mitotic arrest is essential for the normal differentiation of male germ cells into spermatogonia and accompanies epigenetic reprogramming and meiosis inhibition from embryonic development to post-hatch. However, mitotic arrest was not well studied in chickens because of the difficulty in obtaining pure germ cells from relevant developmental stage. Results We performed single-cell RNA sequencing to investigate transcriptional dynamics of male germ cells during mitotic arrest in DAZL::GFP chickens. Using differentially expressed gene analysis and K-means clustering to analyze cells at different developmental stages (E12, E16, and hatch), we found that metabolic and signaling pathways were regulated, and that the epigenome was reprogrammed during mitotic arrest. In particular, we found that histone H3K9 and H3K14 acetylation (by HDAC2) and DNA demethylation (by DNMT3B and HELLS) led to a transcriptionally permissive chromatin state. Furthermore, we found that global DNA demethylation occurred gradually after the onset of mitotic arrest, indicating that the epigenetic-reprogramming schedule of the chicken genome differs from that of the mammalian genome. DNA hypomethylation persisted after hatching, and methylation was slowly re-established 3 weeks later. Conclusions We found a unique epigenetic-reprogramming schedule of mitotic-arrested chicken prospermatogonia and prolonged hypomethylation after hatching. This will provide a foundation for understanding the process of germ-cell epigenetic regulation in several species for which this process is not clearly described. Our findings on the biological processes related to sex-specific differentiation of prospermatogonia could help studying germline development in vitro more elaborately.

show abstract

Section: Methodsmentioning

confidence: 99%

Single-cell RNA sequencing of mitotic-arrested prospermatogonia with DAZL::GFP chickens and revealing unique epigenetic reprogramming of chickens

Choi

Jung

Rengaraj

et al. 2022

J Animal Sci Biotechnol

Self Cite

View full text Add to dashboard Cite

show abstract

“…For PGC-mediated genome editing in avian species, several methods to isolate avian PGCs have been established [130][131][132]. Subsequently, optimal culture conditions for PGC originated from different avian species have to be established.…”

Section: Future Research and Conclusionmentioning

confidence: 99%

Current Approaches and Applications in Avian Genome Editing

Lee

Kim

Lee

2020

IJMS

View full text Add to dashboard Cite

Advances in genome-editing technologies and sequencing of animal genomes enable researchers to generate genome-edited (GE) livestock as valuable animal models that benefit biological researches and biomedical and agricultural industries. As birds are an important species in biology and agriculture, their genome editing has gained significant interest and is mainly performed by using a primordial germ cell (PGC)-mediated method because pronuclear injection is not practical in the avian species. In this method, PGCs can be isolated, cultured, genetically edited in vitro, and injected into a recipient embryo to produce GE offspring. Recently, a couple of GE quail have been generated by using the newly developed adenovirus-mediated method. Without technically required in vitro procedures of the PGC-mediated method, direct injection of adenovirus into the avian blastoderm in the freshly laid eggs resulted in the production of germ-line chimera and GE offspring. As more approaches are available in avian genome editing, avian research in various fields will progress rapidly. In this review, we describe the development of avian genome editing and scientific and industrial applications of GE avian species.

show abstract

“…This characteristic provides them with a huge advantage in germ cell research. Various methods have been used to isolate PGCs from embryonic tissue or blood vessels, including fluorescence-activated cell sorting (FACS), magnetic-activated cell sorting (MACS) using PGC-specific antibodies, and density gradient centrifugation, and size-dependent isolation without using antibodies (Chang et al, 1992;Ono and Machida, 1999;Zhao and Kuwana, 2003;Mozdziak et al, 2005;Jung et al, 2017).…”

Section: Application Of Avian Pgcsmentioning

confidence: 99%

The early development of germ cells in chicken

Kim¹,

Han²

2018

Int. J. Dev. Biol.

View full text Add to dashboard Cite

Primordial germ cells (PGCs) are the founder cells for mature gametes, the vehicles by which individuals transmit genetic and epigenetic information to later generations. Since the 19 century, avian species (chickens in particular) have been widely used for germ cell research. Previous studies have used chicken PGCs for a variety of research applications, including as a model for studies focusing on germline development. Other applications of chicken PGCs, including conservation efforts for avian species and methods of producing transgenic birds, have further reinforced the importance of these cells. However, much remains to be revealed about the origin and role of PGCs during their development in the chicken. Here, we provide a comprehensive review of chicken PGCs, focusing in particular upon their initial profiles and physiological changes during development as regulated by environmental factors and/or intrinsic mechanisms. We also emphasise sex-dependent differences in PGC development after settlement within the gonads, as well as future applications for avian PGCs.

show abstract

Size-dependent isolation of primordial germ cells from avian species

Cited by 15 publications

References 41 publications

Single-cell RNA sequencing of mitotic-arrested prospermatogonia with DAZL::GFP chickens and revealing unique epigenetic reprogramming of chickens

Single-cell RNA sequencing of mitotic-arrested prospermatogonia with DAZL::GFP chickens and revealing unique epigenetic reprogramming of chickens

Current Approaches and Applications in Avian Genome Editing

The early development of germ cells in chicken

Contact Info

Product

Resources

About