The permeability to cytochalasin B of the new unpigmented surface in the first cleavage farrow of the newt ' s egg

Two to 10 /ig/ml cytochalasin B (CB) caused retraction of the first cleavage furrow in Triturus eggs, a spreading of the unpigmented surface from the furrow region and a flattening of the whole egg. CB appears to act against the contractility of the microfilamentous band at mid-cleavage so as to relax the furrow and also to weaken unpigmented surface to allow the egg to flatten. Uncleaved eggs and the initial formation of the cleavage groove were unaffected by CB. A fully-retracted first cleavage furrow reformed itself on transfer of the egg to normal medium but only at the time of second cleavage. Initiation of second cleavage depended upon there being sufficient of the original pigmented surface on the animal hemisphere. Tritium-labelled CB of high specific activity was prepared and used to study its ability to penetrate the surface of newt eggs during cleavage. Scintillation counting of whole eggs showed that CB was not taken into the newt egg until mid-cleavage (about 17 min after the double stripe stage) when new surface began to spread in the cleavage furrow. Fixation in glutaraldehyde and osmium tetroxide retained radioactivity in the egg, but more CB was retained after fixation in osmium tetroxide alone than after double fixation. Most of the retained radioactivity was in yolk platelets. Autoradiographs were prepared of sectioned eggs which had been fixed at late cleavage after [H]CB had flattened the furrow. These showed that CB entered the egg through the unpigmented surface which formed in the furrow but it could not enter through the pigmented surface. The impermeability of the pigmented surface explains the observations that CB does not prevent initial furrowing at cleavage. Once inside the egg CB is transported slowly. CB penetrates to a limited extent beneath the pigmented surface from its border with the unpigmented surface in the first cleavage furrow and this seems insufficient in some circumstances to suppress the contractile phase of second cleavage.