Human Reproduction, Vol 13, 682-689, Copyright © 1998 by Oxford University Press
YZ Ji, JP Wolf, P Jouannet and M Bomsel
Since alpha6beta1 integrin has been shown to function as a sperm adhesion
receptor in the mouse, we investigated the potential role of beta1 integrin
in the gamete fusion process in humans. The expression of beta1 integrin
was morphologically analysed by indirect immunofluorescence and confocal
microscopy. A homogeneous and intense staining was detected at the plasma
membrane, and in some subcortical vesicles of germinal vesicle stage
oocytes (GV). Beta1 almost disappeared from oolemma and cytoplasm of
metaphase I (MI) oocytes, but was re-expressed as asymmetrical patches at
the plasma membrane of metaphase II stage oocytes (MII). A functional
fusion assay based on Hoechst or calcein-AM dye transfer from one gamete to
the other showed that maturing oocytes were able to fuse with an increasing
number of spermatozoa (11-22 from GV to MII respectively), and that fused
spermatozoa co-localized with beta1 integrin patches. Human gamete fusion
was only partially inhibited either by RGD-containing peptide (GRGDTP), or
by blocking anti-human beta1 integrin monoclonal antibody (DE9), with a
maximum of 50% inhibition. Despite the combined addition of GRGDTP and
blocking mouse anti-human beta1 integrin DE9 in the assay, a complete
inhibition of fusion could not be achieved. A mouse polyclonal antibody
raised against human oocyte membranes was more potent in inhibiting the
fusion. Since beta1 integrin expression at the plasma membrane was not
correlated to oocyte fusibility, and since it was only partially inhibited
by DE9 and/or RGD peptide, we suggest that human gamete fusion can bypass
the beta1 requirement. Beta1 integrin certainly participates in human
gamete fusion by acting in co-operation with multiple integrin/disintegrin
couples or another cofactor, not yet identified.
ARTICLES
Human gamete fusion can bypass beta1 integrin requirement
Laboratoire de Biologie de la Reproduction, Hopital Cochin, Paris, France.
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