title: Protein profile of capacitated versus ejaculated human sperm.
abstract: Freshly ejaculated sperm acquire the fertilizing potential by a continuing process that occurs during sperm transport through the female genital tract, and it is physiologically not complete until the spermatozoon reaches the oocyte. The process termed capacitation can be mimicked in vitro by using appropriate capacitation media. Despite its importance, the molecular mechanisms underlying capacitation are poorly understood. This work deals with a proteomic approach to the analysis of protein profile variations in human normospermic samples as a consequence of three hours in vitro capacitation. 2DE gels were produced per freshly ejaculated sperm and per capacitated sperm and several quantitative and qualitative significant variations were found. Among the MS obtained identifications, proteins with a significant decrease after capacitation were found to be involved in protein fate, metabolism, and flagellar organization; on the contrary, increasing proteins were found to be related to cellular stress. Interestingly, the detected flagellar organization proteins decreased during capacitation whereas their corresponding fragments increased. A swim-up selected and three-hour capacitated sperm subpopulation has also been resolved by 2DE, and its synthetic gel has been analyzed for the variations observed in the entire sperm population. An immunofluorescence analysis of this sperm typology was carried out with antiactin and antitubulin antibodies.
authors.affiliation: Laboratory of Cell Biology, Department of Evolutionary Biology, Siena University, Siena, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Secciani
authors.forename: Laura
authors.initials: L
authors.lastname: Bianchi
authors.forename: Leonardo
authors.initials: L
authors.lastname: Ermini
authors.forename: Riccardo
authors.initials: R
authors.lastname: Cianti
authors.forename: Alessandro
authors.initials: A
authors.lastname: Armini
authors.forename: Giovan Battista
authors.initials: GB
authors.lastname: La Sala
authors.forename: Riccardo
authors.initials: R
authors.lastname: Focarelli
authors.forename: Luca
authors.initials: L
authors.lastname: Bini
authors.forename: Floriana
authors.initials: F
authors.lastname: Rosati
mesh_terms: D017209:Apoptosis; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005260:Female; D005306:Fertilization; D006801:Humans; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods; D011506:Proteins / Q000737:chemistry*; D040901:Proteomics / Q000379:methods; D016622:Silver Staining; D013050:Spectrometry, Fluorescence / Q000379:methods; D013075:Sperm Capacitation / Q000502:physiology*; D013094:Spermatozoa / Q000378:metabolism / Q000473:pathology*
chemical_list: D011506:Proteins