Mechanical Properties Measurement,proteomic And Genomic Studies Of Idiopathic Male Infertility On A Chinese Consanguineous Family

Part ? Spotlight on mechanical properties of abnormal shape sperm Mechanical properties,as the inherent characteristics of cells,were recognized in numerous biological processes,and mechanical properties had been recognized as an indicator of pathological modifications of cells.While comparatively in sperm,it was still neglected.As such,we conducted to study mechanical properties of sperm head with normal and abnormal morphology,and investigate the association between mechanical properties and status of chromatin condensation of sperm head.The mechanical properties were investigated by atomic force microscopy(AFM)under physiological conditions.Chromatin condensation was detected by aniline blue staining,western blot,transmission Electron Microscopy Imaging and sperm chromatin structure assay(SCSA).We observed that geometric characteristics,stiffness and the status of chromatin condensation changed evidently in pathological sperm.The results revealed that the increase in length of sperm head was mainly determined by nuclear.Abnormal sperm chromatin condensation might lead to change in stiffness of sperm head.And stiffness of sperm may be a new indicator to assess sperm quality.Additionally,for the first time,the current study provided a new biomechanics strategy for evaluating pathological sperm contributes to our understanding of male infertility.Part ? Proteomic studies of idiopathic male infertility on a Chinese consanguineous family Two patients from a consanguineous family presenting with completely abnormal shape of sperm,and 40 male controls with normal fertility were involved in this study.Sperm proteins were extracted by urea/thiourea and separated by two-dimensional gel electrophoresis(2-DE).After silver staining and image scanning,different expressed proteins were identified by matrix-assisted laser desorption ionization time-of-flight tandem mass spectrometry(MALDI-TOF-MS).Protein function was analyzed by the David database.And the protein-protein interaction network was analyzed by the String database.A total of 26 proteins spots were found differentially expressed,12 showed up regulation while 14 showed down regulation in teratozoospermia compared with control group.The clear 26 differential protein spots were identified by MALDI-TOP-MS.Semenogelin-1,Semenogelin-2,SEMG2 protein were found to be up-regulated in the teratozoospermia group.Ropporin-1A isoform a,Clusterin preproprotein,ASRGL1 protein,Glutathione S-transferase Mu 3,Tubulin alpha-3E chain,Tubulin beta-4B chain,Sperm protein associated with the nucleus on the X chromosome C,SPANX-C,Succinate--Co A ligase [ADP-forming] subunit beta,mitochondrial were down-regulated in the teratozoospermia group.A system biology approach was used to analyze the proteins to major invoved in binding,catalytic activity and structural molecule activity.These results suggested that Semenogelin-1 preproprotein,Semenogelin-2 preproprotein,SEMG2,GSTMu3 may be associated with defects in sperm motility.TUBB4 B,TUBA3E,Ropporin,SPANX-C may be associated with defects in sperm morphology.CLU,HYOU1,SUCLA2 may be associated with quantitative impairment of spermatogenesis in the Chinese consanguineous family.Part ? Genomic studies of idiopathic male infertility on a Chinese consanguineous family Male infertility affects approximately 7% of the male population.In about 40% of the patients the aetiology remains unknown.Here,we identified a novel homozygous nonsynonymous mutation(c.1991C>T/p.Gly664Asp)of FBXO43 in a consanguineous Chinese family using whole-exome sequencing.The recessive mutation was parentally inherited from heterozygous carriers but was absent in 200 fertile individuals.The mutation was segregating with the disease phenotype and was predicted to be a disease causing protein by SIFT,Poly Phen-2 and Mutation Taster.The identified mutation is located in the zinc finger of the FBXO43 protein.An in silico mutant FBXO43 model predicts that the mutation p.Gly664 Asp causes an additional ?-helix and shortens of two ?-sheets,which may resulting in loss of function of the protein.Therefore,these findings support the important role of the FBXO43 mutation in determining male infertility and expand our understanding of the genetic basis of male infertility.