The Expression And Function Of NASP In The Endometrium Of Early Pregnant Mice

Objective: Nuclear auto-antigenic sperm protein(NASP),initially described as a highly auto-immunogenic testis and sperm-specific protein.It is a histone chaperone that can bind histone and transfer them from cytoplasm to cell nucleus,which participant with cell proliferation.This article is to test the the expression and function of NASP in uterus during early pregnancy and better understand the mechanism of embryo implantation.Methods: Tissue collection:(1)mouse normal pregnancy :pregnancy day one(D1)? D4 ? D5 ? D6 ? D8(aginal plug = day 1 of pregnancy);(2)pseudopregnancy: pseudopregnancy day one(PD1)?PD4?PD5?PD6?PD8;(3)artificial induced decidualization(ID): ID and non-ID;(4)Isolation of Uterine Stromal Cells;(5)normal human endometrial tissue samples: proliferative phases?secretory phases?Decidual tissues.Real Time PCR and western blot were used to examine the NASP mRNA and protein expression,respectively.Statistical analyses were performed using a two-sided unpaired Student's t test,one-way analysis of variance with the aid of SPSS 17.0 statistical software.Differences were considered to be significant if p < 0.05.We used immunofluorescence to detect the location of NASP in uterine Stromal Cells.Results: 1.The expression of NASP protein was expressed in uterine luminal and glandular epithelium on D1.NASP localization shifted to the sub-epithelial uterine stroma region on D4.On D5 IS,the expression of NASP expanded to the entire stromal bed.NASP was intensely expressed in the decidual cells and embryonic cells on D6 IS and on D8 IS.The expression of NASP protein level on D6 IS and D8 IS was higher than the level on other day of pregnancy.The expression of NASP mRNA was no significant statistical differences among groups.2.The expression of NASP was located in the luminal epithelium and glandular epithelium on PD1 and shifted to the sub-epithelial uterine stroma region on PD4,whose expressional trends were as same as the trends of normal pregnancy on D1 and D4.On PD5 and PD6,the expression of NASP was also localized among luminal epithelium,glandular epithelium and stromal cells.On PD8,NASP was weakly expressed in the glandular epithelial cells.The NASP protein signal and mRNA signal were comparable among each group.3.NASP protein signal was weakly detected in the luminal epithelium and glandular epithelium in the control horn;and it was highly expressed in the decidualized cells in the oil-induced uterine horn.NASP protein levels of the oil-induced uterine horn were higher than the levels of the control horn.NASP mRNA of the oil-induced uterine horn were comparable with the expression of the uninjected uterine horn.4.NASP localized in nuclei of glandular epithelial and stromal cells in the secretory and proliferative phase respectively.NASP was prominent in decidual cell.NASP protein level in decidua was higher,compared with that in the secretory and proliferative phase.5.NASP was highly expressed in both cell nuclei and cytoplasm in all the stromal cells.NASP protein level was comparable between the decidual group and the control group.Immunofluorescence of NASP mobilized from the cytoplasm to the nucleus in stromal cells on 0h and 24 h.After 48 h decidual stimuli and the number of nuclear location in decidual groups was more than the number in control group.Conclusion: 1.The expression of Ki67 and NASP indicated that NASP might be involved with the cell proliferation.NASP was highly expressed in decidual cells,which gave the point that NASP may be related with uterus decidualization.2.NASP was expressed both in cell nuclei and cytoplasm in all the stromal cells at 0h and 24 h.After undergone artificial decidualization,NASP was mainly expressed in cell nuclei,and the number is more than the number of control group.These results indicated that NASP was related with cell cell proliferation.Objective: Drosophila Eph kinase(DEK)was a proto-oncogene protein and a nuclear phosphoprotein that can be implicated in DNA replication,DNA double-strand break repair,mRNA splicing,and transcriptional regulation This article is to test the the expression and function of DEK in human endometrium decidulization and better understand the mechanism of embryo implantation.Methods: normal human endometrial tissue samples: proliferative phases ?secretory phases?Decidual tissues.RT-PCR?WB and IHS were used to examine the DEK mRNA and protein expression,respectively.We used immunofluorescence to detect the location of PH3.Alkaline Phosphatase Assay tested to cell differentiation.Flow cytometry detected cell cycle and cell apotosis.Results: DEK localized in nuclei of glandular epithelial and stromal cells in the secretory and proliferative phase respectively.DEK was prominent in decidual cell.DEK protein level in decidua was higher,compared with that in the secretory and proliferative phase.human endometrium decidul cells: After treated with DEK si RNA,we found that the expression of IGFBP1?PH3?P53?FANCD2??H2AFX was declined in decidual cells;staining intensity of Alkaline Phosphatase Assay was weak;decidual cells in the G1 phase was increased;total apoptosis were increased significantly.Conclusion: DEK had a key role of human endometrium decidulization.DEK may be implicated in cell proliferation,cell differentiation,cell apoptosis and DNA damage repair to participate in human endometrium decidulization.