Cleavage furrow isolated from newt eggs: Contraction, organization of the actin filaments, and protein components of the furrow

Abstract

The cleavage-furrow region was isolated surgically from newt eggs at the early stage of the first cleavage. The isolated furrow contracted in the presence of ATP at a Ca2+ concentration of 10 or 100 nM, although the speed was less than that of the furrow in vivo. Cytochalasin B, cytochalasin D, phalloidin, p-chloromercuribenzoate, and N-ethylmaleimide interfered with the contraction, but colchicine did not. The furrow contained bundles of actin filaments of opposite polarities oriented parallel to the long axis of the furrow; these bundles may be the main component of the contractile arc. From electron microscopic observation of thin sections of the furrow, it was suggested that the actin bundles of the contractile arc were organized from preexisting cortical filaments that were connected to the plasma membrane by granular materials at their barbed ends. Contractile-arc actin filaments were revealed to be crosslinked by thin strands by the rapid freezing/deep etching-replication technique. Two-dimensional polyacrylamide gel electrophoresis showed that several proteins found in the furrow cortex are absent from the cortical layer before the cleavage furrow is formed.