Effects Of Zinc Deficiency And Zinc Excessiveness On The Development Of Cultured Mouse Embryos

Objective: The aim of this study is to observe the effect of teratogenesis induced by zinc deficiency and zinc excessiveness in vitro on the development of mouse embryos, especially on the development of brain and limb bud, to explore its possible mechanisms, and to provide a better understanding of the mechanism of action of zinc deficiency and zinc excessiveness on the mouse embryo.Methods: The post-implantation whole embryo culture (WEC) system in self-made rotator with discontinuous supplementation of gas mixture was used to explore the effects of zinc deficiency and zinc excessiveness on cultured embryos in vitro. The 8.5-day isolated Kunmin mouse embryos (2-5 somites) at organogenetic stage were co-cultured for 48 hours in immediately centrifugal serum (ICS) with TPEN and zinc(final concentration:24.5-392.0 μmol/L ). To explore the characteristic of developmental toxicity and the mechanisms of zinc deficiency and zinc excessiveness, HE staining and light microscope were used to observe damages on structure of visceral yolk sac (VYS) and cultured embryos.In this study, an in vitro method of mouse limb bud culture was also used. By means of this method, Then the potential influence of zinc on the growth and differentiation of it was studied mouse forelimb bud at 12-day stage were co-cultured with different amounts of zinc for 72 hours. Neubert system was used to determine the degree of morphogenetic differentiation of forelimb buds, and flow cytometry was used to detect limb bud cell cycle and apoptosis.Micromass culture (MC) was used to investigate the effects of deficiency...