title: Primordial Germ Cell Isolation from Xenopus laevis Embryos.
abstract: Primordial germ cells (PGCs) are the precursors to the gametes and have the unique ability to retain full developmental potential. However, the mechanism(s) and gene-network(s) necessary for their proper specification and development are poorly understood. This is due, in part, to the challenges that must be overcome in order to identify and isolate PGCs during critical stages of development. Two distinct mechanisms have been characterized to specify the germ cell lineage in vertebrates: induction and inheritance. Regardless of mechanism, there are common developmental features shared among all vertebrates in forming the germ cell lineage. Xenopus offers several advantages for understanding the molecular mechanisms necessary to establish the germ line. Here, we provide detailed methods for isolating live PGCs at different time points: 1) just after they have segregated from the endodermal lineage, and 2) while they are migrating towards the presumptive gonad. Isolation of PGCs at these critical developmental stages will allow for the investigation of the mechanism(s) and gene-network(s) necessary for their proper specification and development.
authors.affiliation: Department of Cell Biology, University of Miami Miller School of Medicine, 1600 NW 10 Ave, Miami, FL, 33136, USA.
authors.forename: Amanda M
authors.initials: AM
authors.lastname: Butler
authors.affiliation: Department of Cell Biology, University of Miami Miller School of Medicine, 1600 NW 10 Ave, Miami, FL, 33136, USA.
authors.forename: Tristan
authors.initials: T
authors.lastname: Aguero
authors.affiliation: Department of Cell Biology, University of Miami Miller School of Medicine, 1600 NW 10 Ave, Miami, FL, 33136, USA.
authors.forename: Karen M
authors.initials: KM
authors.lastname: Newman
authors.affiliation: Department of Cell Biology, University of Miami Miller School of Medicine, 1600 NW 10 Ave, Miami, FL, 33136, USA. MKing@med.miami.edu.
authors.forename: Mary Lou
authors.initials: ML
authors.lastname: King
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D019070:Cell Lineage; D002465:Cell Movement; D002469:Cell Separation / Q000379:methods*; D004707:Endoderm / Q000166:cytology; D018507:Gene Expression Regulation, Developmental; D005854:Germ Cells / Q000166:cytology* / Q000254:growth & development; D013234:Stem Cells / Q000166:cytology; D014982:Xenopus laevis / Q000196:embryology*
keywords: Germ line; Germ plasm; Maternal-to-zygotic transition (MZT); Migration; Primordial germ cell (PGC); Xenopus