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Updated: Aug 10, 2026
Fluorescence in situ hybridization (FISH) Protocol in Human Sperm
Published on: September 1, 2009
Changes in protamine 1 distribution in human sperm nucleus during in vitro sperm-oocyte interaction: an
D Colleu1, D Lescoat, D Thomas
1Center for Infertility and Assisted Reproductive Technology La Sagesse, University of Rennes, France.
Objective:
To determine whether the process of sperm nuclear destabilization would begin before sperm-oocyte fusion in humans.
Design:
Changes in the distribution of human protamine 1 were investigated in human spermatozoa from the ejaculate, in spermatozoa selected by swim-up or Percoll techniques, and in spermatozoa bound to zona pellucida (ZP) from oocytes that failed to fertilize in an IVF program.
Setting:
Center for Infertility and Assisted Reproductive Technology, and university departments.
Patient(S):
Fifteen couples undergoing an IVF program.
Intervention(S):
Women underwent a similar superovulation induction protocol that consisted of GnRH agonist associated with hMG.
Main Outcome Measure(S):
Comparative immunoelectron microscopic study of sperm nucleus labeling with an anti-human protamine 1 specific protamine monoclonal antibody.
Result(S):
After selection by swim-up or by Percoll, spermatozoa show a significantly lower nuclear labeling than in the ejaculate. After binding to the ZP, labeling increases, more in spermatozoa selected by swim-up than by Percoll, but, after Percoll selection, labeling in zona-bound spermatozoa is lower than in the ejaculate.
Conclusion(S):
In humans sperm binding to the ZP induces differences in the accessibility of the anti-human protamine 1 antibody, which are consistent with structural rearrangements of the DNA-nucleoproteins complex. These modifications must be a prelude to sperm decondensation, protamines replacement by histones, and subsequent reactivation of the sperm genome in the oocyte.
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