The Research Of The Crb3Affect The Leydig Cells Of Mouse Secret Testosterone

Object：We detect whether Crb3is expressed in leydig and Complete the celltransfection by plasmid if Crb3exist in leydig Detect the expression of RNA andtestosterone secreted by leydig after the Crb3is overpression by us. The purpose of usis probing the relation of Crb3and the function of testis for secreting testosterone andproviding wizard for the diagnosing and treatment of TDS.Methods：[1] Detect the expression of Crb3in leydig cell with the method of PCRWe extract the Total RNA from the sub-culture cell of leydig cell and reversetranscript RNA to C-DNA with Reverse transcription kit. Acquire the target DNAfrom the C-DNAby normal PCR。2、Detect the translation of Crb3protein in leydig with the method ofWestern-blotWe extract the total protein from sub-culture cell of leydig and detect whetherCrb3translate to protein and the lever of translation.3、Complete the cell transfection by plasmid.We transfect plasmid which over-express Crb3of leydig cell into leydig cell anddetect the change of Crb3RNAexpression by RT-PCR and the change of testosteronesecreted by leydig by Chemiluminescence. The experiment of transfection is dividedinto three groups which are group of transfection、no-tranfection and emptytransfection.4、statistic analysisWe build the experiment data and analysis it with the software of spss19.Theanalysis of variance is used to analysis the average of data. The meaning of statistic isexist for P<0.5.Result：1、The DNAexpression and meaning of Crb3in leydig cellThe conclusion of normal PCR shown that Crb3express lowly in leydig cell.Consider the research of Crb3before that Crb3can affect the function of epithelial,then we wounder whether the Crb3would affect the function of leydigcell.2、The protein translation and meaning of Crb3in leydigcellThe result of western-blot show that the protein translation present low lever inleydig cell.The phenomenon just mach the DNAexpression of Crb3.3、The DNAexpression of Crb3of group of tranfection is higher than the groupof no-tranfection and empty transfection.And the meaning of statistic is exist forP<0.5.4、The result of transfection show that the protein expression of is lower thanthe group of no-tranfection and empty transfection.And the meaning of statistic isexist for P<0.5.Conclusion：The over-expression of Crb3can decrease the testosterone which issecreted by the leydig cells of mouse.