title: Deleted in azoospermia-like enhances in vitro derived porcine germ cell formation and meiosis.
abstract: Evidence supporting that deleted in azoospermia-like (DAZL) plays a key role during gametogenesis and meiosis continues to emerge. Our study aimed to determine whether overexpression of DAZL using a lentiviral approach in a somatic stem cell to germ cell in vitro differentiation culture could enhance the formation of primordial germ cell-like cells (PLCs) and oocyte-like cells (OLCs). Introduction of DAZL at the beginning of induced differentiation significantly increased the formation of Fragilis-positive PLCs, which was independent of mitotic proliferation. In addition, mRNA levels of the germ cell markers Oct4, Stella, and Vasa were also higher in the DAZL-transduced group and suppressed when DAZL was knocked down using small interference RNA. At later stages of differentiation, the expression of several genes associated with meiosis, including Scp3, Dmc1, Rec8, and Stra8, was determined to be significantly higher when DAZL was overexpressed, which was abrogated by its knockdown. Exogenous introduction of DAZL also increased the protein levels of SCP3 and VASA, which again was reversed by its knockdown. Although not a common phenomenon in the in vitro differentiation system, the percentage of SCP3-positive cells displaying meiotic chromosome patterns in the DAZL-transduced group was higher than in the control, as was the overall percentage of OLCs that were generated. The introduction of factors such as DAZL into a stem cell-to-germ cell differentiation culture may provide an opportunity to better understand the key genes and their interactions during gametogenesis, also providing a means to enhance the generation of germ cells in vitro.
authors.affiliation: Department of Animal and Poultry Science, University of Guelph, Guelph, Ontario, Canada.
authors.forename: Bong-Wook
authors.initials: BW
authors.lastname: Park
authors.forename: Wei
authors.initials: W
authors.lastname: Shen
authors.forename: Katja
authors.initials: K
authors.lastname: Linher-Melville
authors.forename: Julang
authors.initials: J
authors.lastname: Li
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D002454:Cell Differentiation; D002467:Cell Nucleus / Q000378:metabolism; D048430:Cell Shape; D002478:Cells, Cultured; D004261:DNA Replication; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D005854:Germ Cells / Q000502:physiology*; D008540:Meiosis; D034741:RNA, Small Interfering / Q000235:genetics; D016601:RNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology*; D012867:Skin / Q000166:cytology; D013234:Stem Cells / Q000502:physiology; D034421:Sus scrofa
chemical_list: D000943:Antigens, Differentiation; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins