2098 REMODELING OF HUMAN SPERM CHROMATIN DURING IN VITRO CAPACITATION AND ACROSOME REACTION: EVIDENCE FROM CYTOCHEMICAL TESTS AND CORRELATIONS BETWEEN ASSAYS

Abstract

You have accessJournal of UrologyInfertility: Physiology, Pathophysiology, Basic Research1 Apr 20122098 REMODELING OF HUMAN SPERM CHROMATIN DURING IN VITRO CAPACITATION AND ACROSOME REACTION: EVIDENCE FROM CYTOCHEMICAL TESTS AND CORRELATIONS BETWEEN ASSAYS Eve de Lamirande, Maria San Gabriel, Naif Alhathal, and Armand Zini Eve de LamirandeEve de Lamirande Montreal, Canada More articles by this author , Maria San GabrielMaria San Gabriel Montreal, Canada More articles by this author , Naif AlhathalNaif Alhathal Montreal, Canada More articles by this author , and Armand ZiniArmand Zini Montreal, Canada More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.2265AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Capacitation (CAP) and acrosome reaction (AR), are sequential processes of sperm activation. Beside the known ionic, membrane and transduction events and final release of proteolytic enzymes to help sperm movement towards the egg, changes to chromatin are also expected. These should involve remodeling rather than DNA damage. Our aims were evaluate the changes in human sperm head using cytochemical stains and to confirm that CAP/AR do not include DNA damage. METHODS Percoll-washed spermatozoa were incubated in BWW +/− fetal cord serum ultrafiltrate (10%;3.5h;CAP) and, subsequently, with lysophosphatidylcholine (2.5 μM; 30 min, AR). Then, ethanol-fixed spermatozoa were used for: PSA-FITC (for AR), aniline blue (AB, for histones), chromomycin A3 (CMA3, for protamines), toluidine blue (TB, for chromatin compaction), iodoacetamide-fluorescein (IAF, for sulfhydryl groups), induced decondensation, sperm chromatin structure assay (DNA fragmentation index, DFI) and immunoblotting (for histone, H2B). RESULTS CAP/AR was associated with similar increases in staining for AB (∼75%) and TB (∼50%) but had no inflence on CMA3. The increase (∼55%) in IAF staining observed during CAP was abscent after AR. CAP/AR did not damage DNA (DFI remained low) nor affect histone content. CAP, and even more AR, primed sperm heads to decondense (∼100% and ∼160% increases, respectively) when challenged with SDS (1%) + DTT (0.1 mM). Interestingly, induced decondensation correlated with all other tests CAP, AB, TB, IAF; also CAP was related to AB and TB stains. CONCLUSIONS The data strongly support human sperm chromatin remodeling during CAP/AR, these modifications are interlinked and probably help prepare chromatin for after-fertilization events. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e845-e846 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Eve de Lamirande Montreal, Canada More articles by this author Maria San Gabriel Montreal, Canada More articles by this author Naif Alhathal Montreal, Canada More articles by this author Armand Zini Montreal, Canada More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...