Characterization of Oct4-GFP transgenic mice as a model to study the effect of environmental estrogens on the maturation of male germ cells by using flow cytometry

• Published in: The Journal of steroid biochemistry and molecular biology 2015-11, Vol.154, p.53-61
• Main Authors: Porro, Valentina, Pagotto, Romina, Harreguy, María Belén, Ramírez, Sofía, Crispo, Martina, Santamaría, Clarisa, Luque, Enrique H., Rodríguez, Horacio A., Bollati-Fogolín, Mariela
• Format: Article
• Language: English
• Subjects: Animals; Environmental Pollutants - pharmacology; Estrogens; Ethinyl Estradiol - pharmacology; Flow cytometry; Gene Expression Profiling; GFP; Green Fluorescent Proteins - genetics; Male; Male germ cells; Maturation; Mice; Mice, Inbred C57BL; Mice, Transgenic; Oct4; Octamer Transcription Factor-3 - genetics; Octamer Transcription Factor-3 - physiology; Sperm Count; Sperm Motility; Spermatozoa - drug effects
• ISSN: 0960-0760; 1879-1220
• DOI: 10.1016/j.jsbmb.2015.06.006

•We study changes in male germ cells by exposure to xenoestrogens in Oct4/GFP mouse.•Oct4/GFP maturation profile from neonatal period until adult stage was established.•Neonatal Oct4 expression increased after perinatal exposure to ethynilestradiol.•Sperm count in adult mice was reduced after perinatal exposure to ethynilestradiol.•Oct4/GFP mouse is useful to detect changes in male germ cells by xenoestrogens. Oct4 is involved in regulation of pluripotency during normal development and is down-regulated during formation of postnatal reservoir of germ cells. We propose thatOct4/GFP transgenic mouse, which mimics the endogenous expression pattern of Oct4, could be used as a mammalian model to study the effects of environmental estrogens on the development of male germ cells. Oct4/GFP maturation profile was assessed during postnatal days -PND- 3, 5, 7, 10, 14 and 80, using flow cytometry. Then, we exposed pregnant mothers to 17α-ethinylestradiol (EE2) from day post coitum (dpc) 5 to PND7. Percentage of Oct4/GFP-expressing cells and levels of expression of Oct4/GPF were increased in PND7 after EE2 exposure. These observations were confirmed by analysis of GFP and endogenous Oct4 protein in the seminiferous tubules and by a reduction in epididymal sperm count in adult mice. We introduced Oct4/GFP mouse together with flow cytometry as a tool to evaluate changes in male germ cells development.