Effects Of ZSWIM3 Deficiency On Early Embryonic Development In Mice

BackgroundThe development of a mammalian embryo begins with the union of sperm and egg to form a zygote,which produces a pluripotent mass of cells through a series of genetic and epigenetic controls.Early embryonic development is driven by maternal proteins and gene transcripts stored in the egg,and a large number of maternal regulatory factors play a key role in this process.During the maternal-zygote transition(MZT),developmental control is transferred from the mother to the zygote genome with the degradation of maternal transcripts and proteins.The development of embryos after ZGA is controlled by maternal-factor to zygote genome.Thereafter,the zygote genome controls the development of cell lineages during cell differentiation.Maternal-effector genes are activated,driven and transcribed during oocytopenesis,and these factors accumulate during oocytopenesis,enabling embryonic genome activation(ZGA)cleavage and the initial establishment of embryonic cell lines.Up to now,more than 30 mouse genes with maternally mutated effects have been reported.Their functions involve the fusion of male and female prokaryotes,elimination of maternally derived products,activation of zygote genomes,cleavage,and densification of embryos,which have important effects on oogenesis,meiotic maturation,pre-implantation and post-implantation embryonic development.However,the role of most maternal and zygotic genes in embryonic development is still uncertain.ZSWIM3 is refers to a novel zinc finger chelating domain,is also a protein-coding gene that is expected to play.a role in DNA binding and protein-binding interactions.ZSWIM3 is expressed in cerebral cortex,liver,kidney,uterus,testis and other tissues as well as various mesenchymal cells,which proves that it plays an important role in regulating the development of various tissues and organs.It is also expressed in ovarian spermatogonia and spermatocyte.In addition,it has been predicted to be an adverse prognostic marker for breast and endometrial cancer.C57BL/6N is one of the most widely used strains in mice,and is also the most commonly used female parent of transgenic or gene knockout mice in genetic engineering,which can be used as the experimental animal model in this study.Currently,studies on ZSWIM3 mainly focus on alcoholic liver disease and inhibition of inflammatory response,and there are few reports on the expression characteristics and mechanism of ZSWIM3 in mouse oocytes and early embryonic development.In this study,we investigated the expression,distribution and functional mechanism of ZSWIM3 in early embryogenesis of mice.ObjectiveThe localization and distribution of maternal factor ZSWIM3 in the process of embryo development were determined.To explore the effect of ZSWIM3 gene deletion on early embryonic development.To elucidate the role of ZSWIM3 in early embryonic zygotic genomic activation and development.To analyze the regulation mechanism of Zswim3 deletion on upstream and downstream genes.MethodsUsing the mouse egg and embryo acquisition technology,the embryos in each stage of early embryo development required by the experiment were obtained.The localization and distribution of ZSWIM3 in early embryonic development were obtained by immunofluorescence assay.Morpholino technology was used to block the transcription and translation of target genes and knock down the protein expressed by target genes in embryos.Embryos of the treated group and the untreated group were cultured in vitro,and phenotypic changes and development rates of the two groups of embryos were observed by photographing.At the same time,immunofluorescence was used again to detect the protein expression in the Morpholino treated embryos,and EU staining was used to observe the expression of newborn RNA.?-amanitin inhibition test and treatment group compared the protein and newborn RNA.Single cell transcriptome sequencing was performed on embryos from the treatment group and the control group to explore the changing rules of upstream and downstream regulatory factors of target genes and the relevant signaling pathways.Results1.Zswim3 mRNA accumulates during the early development of human and mouse oocytes and suddenly drops to a low level during zygotic genomic activation.2.Zswim3 was widely present in the nucleus of early embryonic mice at all stages of development,and the fluorescence signal was strongest at the 2-cell stage.3.The knockdown of Zswim3 resulted in reduced protein and RNA expression,and the deletion of this gene resulted in embryonic development arrest,mostly at the 2-4cell stage,which proved the importance of Zswim3 in early embryonic development of mice.4.ZSWIM3 knock can lead to lower 583 differences in gene expression,including 59 gene expression,gene expression by 524.5.These differentially expressed genes are mainly involved in biological processes such as cell metabolism,organic metabolism,biological regulation,stress response,protein binding,and gene expression,and are mainly involved in signaling pathways such as transport and catabolism,protein processing,and RNA transport.ConclusionsThis study revealed that Zswim3 was widely present in all stages of early embryonic development in mice,and the deletion of this gene would delay the development of early embryonic mice,mostly in the 2-4 cell stage,and the expression of Zswim3-deficient embryonic protein and newborn RNA was decreased.Biological information analysis indicated that Zswim3 loss would affect the expression of related genes,but further verification is needed.This gene plays an important role in the early embryonic development of mice.