Author:
Boyle Judith A.,Chen Hui,Bamburg James R.
Abstract
Scanning and transmission electron microscopy were used to determine the
morphological changes in the egg plasma membrane associated with sperm
binding, fusion and incorporation in Xenopus laevis. Sperm incorporation
in Xenopus is rapid, occurring within 3-5 min following addition of
sperm. Images have been obtained of both early sperm-egg interactions
and fertilisation bodies. Additionally, two drugs that specifically
alter F-actin dynamics, latrunculin and jasplakinolide, were used to
determine whether sperm incorporation is a microfilament-dependent
process. Jasplakinolide did not prevent sperm incorporation, cortical
granule exocytosis or cortical contraction, suggesting these events
can occur without depolymerisation of existing, stabilised
filaments. Latrunculin A, which competes with thymosin β4 in ooplasm for
binding actin monomer, did not inhibit cortical granule exocytosis, but
blocked cortical contraction in 100% of eggs at a concentration of
5 μM. Although a single penetrating sperm was found on an egg pretreated
in latrunculin, fertilisation bodies were never observed. At <5 μM
latrunculin, many eggs did undergo cortical contraction with some
exhibiting severe distortions of the plasma membrane and abnormal
accumulations of pigment granules. Preincubation of eggs in jasplakinolide
before latrunculin mitigated both these effects to some degree. However,
eggs incubated in latrunculin either prior to or after insemination never
progressed through first cleavage.
Publisher
Cambridge University Press (CUP)
Subject
Cell Biology,Developmental Biology
Cited by
12 articles.
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