Acrosin and tubulin are structural components of the spermatozoon involved in zona pellucida digestion and tail movment respectively. Their cellular localization can be achieved easily by indirect immunofluorescence. In a previous study using polyclonal antibodies they were shown to be involved in the fertilization process. The purpose of this study was to correlate the indirect immunofluorescence and morphology results with the fertilization rates obtained after regular IVF or ICSI and to provide biologists with a quick and efficient tool for the selection of the most appropriate fertilization technique. A 5 ml aliquot from a washed sperm suspension used for regular IVF or ICSI was smerred into a microscopic slide.Monoclonal antibodies against pro-acrosin or tubulin were used for indirect immunofluorescence assessment of the sperm acrosome and tail in a double labelling assay. Double immunofluorescent labbelling of pro-acrosin and tubulin allowed the easy identification of both acrosomal and tail defects in a single reading. The indirecte immunofluorescence procedure described can be easily implemented in a busy IVF unit. It provides clinicians and patients with more detailed descriptions of sperm abnormalites. In cases of abnornal spermatozoo it helps biologist to chose rapidly which fertilization tecnique should be employed
Double fluorescenze labelling of acrosin and tubulin in human spermatozoa: a rapid diagnostic procedure to identify sperm samples with poor fertilizing ability.
MARCI, Roberto;
1997
Abstract
Acrosin and tubulin are structural components of the spermatozoon involved in zona pellucida digestion and tail movment respectively. Their cellular localization can be achieved easily by indirect immunofluorescence. In a previous study using polyclonal antibodies they were shown to be involved in the fertilization process. The purpose of this study was to correlate the indirect immunofluorescence and morphology results with the fertilization rates obtained after regular IVF or ICSI and to provide biologists with a quick and efficient tool for the selection of the most appropriate fertilization technique. A 5 ml aliquot from a washed sperm suspension used for regular IVF or ICSI was smerred into a microscopic slide.Monoclonal antibodies against pro-acrosin or tubulin were used for indirect immunofluorescence assessment of the sperm acrosome and tail in a double labelling assay. Double immunofluorescent labbelling of pro-acrosin and tubulin allowed the easy identification of both acrosomal and tail defects in a single reading. The indirecte immunofluorescence procedure described can be easily implemented in a busy IVF unit. It provides clinicians and patients with more detailed descriptions of sperm abnormalites. In cases of abnornal spermatozoo it helps biologist to chose rapidly which fertilization tecnique should be employedI documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
