COMPARATIVE STUDY OF TWO STAINING TECHNIQUES TO EVALUATE THE ACROSOMAL REACTION IN HUMAN SPERM

POBLETE S; ARIAS RJ; FORNES ML; JUSTRIBÓ G; MARTINEZ A; PELLETÁN LE; SOSA CM; DE BLAS GA

Mendoza

Congreso; XXXIV Reunión Científica Anual de la Sociedad de Biología de Cuyo; 2016

Sociedad de Biología de Cuyo

<!-- /* Font Definitions */@font-face{font-family:Calibri;panose-1:2 15 5 2 2 2 4 3 2 4;mso-font-charset:0;mso-generic-font-family:auto;mso-font-pitch:variable;mso-font-signature:-520092929 1073786111 9 0 415 0;} /* Style Definitions */p.MsoNormal, li.MsoNormal, div.MsoNormal{mso-style-unhide:no;mso-style-qformat:yes;mso-style-parent:"";margin-top:0cm;margin-right:0cm;margin-bottom:8.0pt;margin-left:0cm;line-height:107%;mso-pagination:widow-orphan;font-size:11.0pt;font-family:Calibri;mso-fareast-font-family:Calibri;mso-bidi-font-family:"Times New Roman";mso-fareast-language:EN-US;}.MsoChpDefault{mso-style-type:export-only;mso-default-props:yes;font-size:10.0pt;mso-ansi-font-size:10.0pt;mso-bidi-font-size:10.0pt;font-family:Calibri;mso-ascii-font-family:Calibri;mso-fareast-font-family:Calibri;mso-hansi-font-family:Calibri;}@page WordSection1{size:612.0pt 792.0pt;margin:70.85pt 3.0cm 70.85pt 3.0cm;mso-header-margin:36.0pt;mso-footer-margin:36.0pt;mso-paper-source:0;}div.WordSection1{page:WordSection1;}-->The acrosome is asecretory vesicle located in the anterior region of the sperm head. The acrosomereaction (AR) is an exocytic event triggered by the fusion between the plasmamembrane and the outer acrosomal membrane, with the subsequent release of the acrosomalcontent. The AR is an important marker for human sperm function. Differentlaboratory techniques may be use for the detection of this exocytic process.The aim of this study was to compare two methods (FITC-coupled Pisum sativumagglutinin -FITC-PSA- and Coomassie brilliant blue -CBB-) to evaluate theacrosomal status of human sperm. We worked with normal, cryopreserved andteratozoospermic human semen samples. Capacitated human sperm were stimulatedfor 30 minutes with calcium ionophore (A23187) and progesterone (Pg) andacrosomal status was evaluated using an upright Nikon microscope equipped withepifluorescence optics. We found that similar results wereachieved with FITC-PSA and CBB staining. These findings indicate that CBB isequally effective in determining the acrosomal status in human spermatozoa.Hence, CBB is a fast, simple and economical method that can be implemented inlow complexity clinical and reproduction laboratories. Thematic area: Developmental Biologyand Reproduction. Mendoza