Characterisation of the deleted in azoospermia like (Dazl)–green fluorescent protein mouse model generated by a two-step embryonic stem cell-based strategy to identify pluripotent and germ cells
Priscila Ramos-Ibeas A B , Eva Pericuesta A , Raúl Fernández-González A , Alfonso Gutiérrez-Adán A and Miguel Ángel Ramírez A
+ Author Affiliations
- Author Affiliations
A Departamento de Reproducción Animal, INIA, Avenida Puerta de Hierro N. 12, local 10, 28040 Madrid, Spain.
B Corresponding author. Email: priscilaramosibeas@gmail.com
Reproduction, Fertility and Development 28(11) 1741-1752 https://doi.org/10.1071/RD14253
Submitted: 17 July 2014 Accepted: 4 April 2015 Published: 6 May 2015
Abstract
The deleted in azoospermia like (Dazl) gene is preferentially expressed in germ cells; however, recent studies indicate that it may have pluripotency-related functions. We generated Dazl–green fluorescent protein (GFP) transgenic mice and assayed the ability of Dazl-driven GFP to mark preimplantation embryo development, fetal, neonatal and adult tissues, and in vitro differentiation from embryonic stem cells (ESCs) to embryoid bodies (EBs) and to primordial germ cell (PGC)-like cells. The Dazl-GFP mice were generated by a two-step ESC-based strategy, which enabled primary and secondary screening of stably transfected clones before embryo injection. During preimplantation embryo stages, GFP was detected from the zygote to blastocyst stage. At Embryonic Day (E) 12.5, GFP was expressed in gonadal ridges and in neonatal gonads of both sexes. In adult mice, GFP expression was found during spermatogenesis from spermatogonia to elongating spermatids and in the cytoplasm of oocytes. However, GFP mRNA was also detected in other tissues harbouring multipotent cells, such as the intestine and bone marrow. Fluorescence was maintained along in vitro Dazl-GFP ESC differentiation to EBs, and in PGC-like cells. In addition to its largely known function in germ cell development, Dazl could have an additional role in pluripotency, supporting these transgenic mice as a valuable tool for the prospective identification of stem cells from several tissues.
Additional keywords: differentiation, oogenesis, pluripotency, primordial germ cells, spermatogenesis, transgenesis.
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