Study of endometrial epithelial-mesenchymal transition of peri-implantation period based on mouse lineage tracing
Objective:To find the direct morphological evidence of endometrial epithelial-mesenchymal transition(EMT)of peri-implantation period by lineage tracing with LtfcreRosa26mT/mG mouse model.Methods:Frozen sections of the uterine tissue from 8-week-old female Ltfcre Rosa26mT/mG mice were collected and co-stained with E-cadherin,Vimentin and CD45,markers of epithelial cells,mesenchymal cells and immune cells,respectively,and were observed under laser scanning confocal fluorescence microscopy.The peri-implantation study was conducted by collecting the uterus of 8-week-old female LtfcreRosa26mT/mG mice at 2.5 dpc,3.5 dpc,4.0 dpc and 4.5 dpc to study the effect of embryo implantation on endometrial EMT.The estrogen-progesterone sequential stimulation model was established using 8-week-old female LtfcreRosa26mT/mG mice which received ovariectomy and sequential estrogen and progesterone injection,and were sacrificed to help explore the effects of hormones on endometrial EMT.Results:Scattered distribution of enhanced green fluorescent protein(EGFP+)cells in the endometrial stroma of LtfcreRosa26mT mG mice was found.After co-staining with E-cadherin,Vimentin and CD45,laser scanning confocal fluorescence microscopy showed part of the scattered EGFP+cells in stroma were Vimentin+stromal cells,namely stromal cells formed by epithelial cells EMT.The results of peri-implantation study showed that the number of EGFP+cells of 3.5dpc was significantly lower than those of dpc2.5 and dpc4.0 during peri-implantation period(P＜0.01).In the establishing estrogen-progesterone sequential stimulation model,the number of EGFP cells in stroma was significantly decreased after estradiol(E2)stimulation,while significantly increased after 2 days of rest,and significantly decreased again after progesterone(P4)stimulation.Conclusions:There are stromal cells formed by epithelial cells EMT in endometrium.EMT already exists in non-embryo implantation period and is strictly regulated by hormones.
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