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and
*Division of Reproductive Sciences, Oregon Regional Primate
Research Center and Departments of Cell and Developmental Biology, and
Obstetrics and Gynecology, Oregon Health Sciences University, Portland,
Oregon 97006; and
To explore the role of nonmuscle myosin II isoforms during
mouse gametogenesis, fertilization, and early development, localization and microinjection studies were performed using monospecific antibodies to myosin IIA and IIB isotypes. Each myosin II antibody recognizes a
205-kDa protein in oocytes, but not mature sperm. Myosin IIA and IIB
demonstrate differential expression during meiotic maturation and
following fertilization: only the IIA isoform detects metaphase spindles or accumulates in the mitotic cleavage furrow. In the unfertilized oocyte, both myosin isoforms are polarized in the cortex
directly overlying the metaphase-arrested second meiotic spindle.
Cortical polarization is altered after spindle disassembly with
Colcemid: the scattered meiotic chromosomes initiate myosin IIA and
microfilament assemble in the vicinity of each chromosome mass. During
sperm incorporation, both myosin II isotypes concentrate in the second
polar body cleavage furrow and the sperm incorporation cone. In
functional experiments, the microinjection of myosin IIA antibody
disrupts meiotic maturation to metaphase II arrest, probably through
depletion of spindle-associated myosin IIA protein and antibody binding
to chromosome surfaces. Conversely, the microinjection of myosin IIB
antibody blocks microfilament-directed chromosome scattering in
Colcemid-treated mature oocytes, suggesting a role in mediating
chromosome-cortical actomyosin interactions. Neither myosin II
antibody, alone or coinjected, blocks second polar body formation, in
vitro fertilization, or cytokinesis. Finally, microinjection of a
nonphosphorylatable 20-kDa regulatory myosin light chain specifically
blocks sperm incorporation cone disassembly and impedes cell cycle
progression, suggesting that interference with myosin II
phosphorylation influences fertilization. Thus, conventional myosins
break cortical symmetry in oocytes by participating in eccentric
meiotic spindle positioning, sperm incorporation cone dynamics, and
cytokinesis. Although murine sperm do not express myosin II, different
myosin II isotypes may have distinct roles during early embryonic
development.
Department of Physiology and
Biophysics, University of Illinois at Chicago, Chicago, Illinois 60612
Corresponding author: Oregon Regional
Primate Research Center and Oregon Health Science University, 505 N.W.
185th Avenue, Beaverton, OR 97006-3499. E-mail: schatten{at}ohsu.edu.
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