title: Measuring intracellular Ca2+ changes in human sperm using four techniques: conventional fluorometry, stopped flow fluorometry, flow cytometry and single cell imaging.
abstract: Spermatozoa are male reproductive cells especially designed to reach, recognize and fuse with the egg. To perform these tasks, sperm cells must be prepared to face a constantly changing environment and to overcome several physical barriers. Being in essence transcriptionally and translationally silent, these motile cells rely profoundly on diverse signaling mechanisms to orient themselves and swim in a directed fashion, and to contend with challenging environmental conditions during their journey to find the egg. In particular, Ca(2+)-mediated signaling is pivotal for several sperm functions: activation of motility, capacitation (a complex process that prepares sperm for the acrosome reaction) and the acrosome reaction (an exocytotic event that allows sperm-egg fusion). The use of fluorescent dyes to track intracellular fluctuations of this ion is of remarkable importance due to their ease of application, sensitivity, and versatility of detection. Using one single dye-loading protocol we utilize four different fluorometric techniques to monitor sperm Ca(2+) dynamics. Each technique provides distinct information that enables spatial and/or temporal resolution, generating data both at single cell and cell population levels.
authors.affiliation: Departamento de Genética del Desarrollo y Fisiología Molecular, Instituto de Biotecnología-Universidad Nacional Autónoma de México.
authors.forename: Esperanza
authors.initials: E
authors.lastname: Mata-Martínez
authors.forename: Omar
authors.initials: O
authors.lastname: José
authors.forename: Paulina
authors.initials: P
authors.lastname: Torres-Rodríguez
authors.forename: Alejandra
authors.initials: A
authors.lastname: Solís-López
authors.forename: Ana A
authors.initials: AA
authors.lastname: Sánchez-Tusie
authors.forename: Yoloxochitl
authors.initials: Y
authors.lastname: Sánchez-Guevara
authors.forename: Marcela B
authors.initials: MB
authors.lastname: Treviño
authors.forename: Claudia L
authors.initials: CL
authors.lastname: Treviño
mesh_terms: D000814:Aniline Compounds / Q000737:chemistry*; D002118:Calcium / Q000032:analysis* / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D005470:Fluorometry / Q000379:methods*; D006801:Humans; D008297:Male; D059010:Single-Cell Analysis / Q000379:methods*; D013094:Spermatozoa / Q000737:chemistry* / Q000378:metabolism; D014966:Xanthenes / Q000737:chemistry*
chemical_list: D000814:Aniline Compounds; D005456:Fluorescent Dyes; D014966:Xanthenes; C059715:Fluo-3; D002118:Calcium