Expression And Localization Of Kinesin KIF2A In Human Sperm

Background and Objective:Through interactions with microtubules,the kinesin-like family member 2A(KIF2A)participates in cell activity,such as transport of various intracellular material,cell migration,cell morphological changes and mitotic spindle microtubules dynamics.Studies have found that KIF2A plays significant roles in neurite growth of neurons and chromosome movement during mitosis by microtubule depolymerization.Recent years it has been reported that the kinesin-like family members have very important impact on spermatogenesis and fertilization of mammalian sperm.Hence,the purpose of this study is to explore the significance of Kif2a in ejaculate of man,and to explore the cellular location of KIF2A protein.Methods:1.Human ejaculates samples were obtained from 100 subjects,sperm RNA was extracted from ejaculate cells by using TRIzol reagent after liquefaction and centrifugation,the RNA was used for reverse transcription into cDNA.The subjects were grouped as abnormal and normal based on the clinical diagnosis of the ejaculate.The amount of Kif2a mRNA was quantified by utilizing real-time quantitative PCR(RT-PCR),Gapdh(glyceraldehyde-3-phosphate dehydrogenase)was selected as housekeeping gene.By use of SPSS 2.0 software to process the data of Kif2a mRNA expression levels in the ejaculate.2.By using immunofluorescence technique to detect KIF2A positioning in sperm.Results:1The results of real-time quantitative PCR showed that the Kif2a mRNA is present in human ejaculates.Conjunction with clinical data of sperm vitality,concentration and total motility(progressive+nonprogressive),we noticed that the expression of Kif2a mRNA in abnormal groups was lower than normal groups,and there were statistically significance between the groups(P<0.(05).2.Immunofluorescence detection shows that the KIF2A located in sperm acrosome region.