Expression Of Luman In Mouse Ovary, Testis And Uterus

Luman is a novel identified protein, which is related to ERS (Endoplasmic ReticulumStress). Luman is one of basic domain zipper transcription factors. Luman in mice isclosely-related to drosophila dCREB-A/BBF-2, dCREB-A functions towards the end ofembryogenesis. Furthermore, Luman was detected in the mouse uterus, ovary and testis. Butthe mechanism and the accurate function of Luman in reproduction were unknown. Theexpression patterns of Luman in the mouse were checked by Real time PCR,immunohistochemistry and Western blot during different time and germinal tissue, includingovary, oviduct and uterus during estrous cycle; Peri-implantation mouse uteri. The biologicalfunctions of Luman were revealed through these studies.1. E.coli with Luman-N-term Fusion gene was induced by IPTG, and identified bySDS-PAGE.The expressed fusion protein was purified by electrical elution. The purifiedLuman-N-term fusion protein was used to immune BALB/C mice. The immunitied micespleen cells and SP2/0myeloma cells were fused under the working PEG(4000), indirectlyELISA method was used to select positive clone and positive clone was screened by limitingdilution method.Finally one hybridoma cell(2C8) which can stably secretinganti-Luamn-N-term protein monoclonal antibodies were obtained. The monoclonal antibodieswere identified: valence of antibody resist Luman-N-term Protein are1×107in abdominaldropsy, the subtypes of monoclonal Antibody is IgG2b, affinity constant of monoclonalAntibody from2C8is1×107.Western-blot indicated that this strain McAb presented highspecificity to Luman-N-term protein expressed in the prokaryotic cells and low specificity toLRF-N-term protein. It is proved that this monoclonal antibody possess well specificity andassociativity, Which were foundation for the further study on function of Luman.2. The expression patterns of Luman in the ovary, oviduct and uterus during estrouscycle were checked by Real time PCR, immunohistochemistry. The expression of Lumanremarkably changed at different stages during estrous cycle, Luman was higher at preoestrusand estrous than others, but lower at dioestrus. This results suggested that Luman might playimportant roles in the female mouse reproduction and the biological functions of Lumanmight be regulated by steroid hormone. 3. The expression of Luman in the peri-implantation pregnant mouse uterus,pseudopregnant mouse uterus, delayed implantation uterus and activated uterus, artificalinduced decidual uterus were checked by Real time PCR, immunohistochemistry and relatedmouse model. These data indicated that Luman might play an important role in embryoimplantation and decidualization and Luman expression is regulated by estrogen and thepresence of an implanted embryo.