Localization and quantification of cyclin A and B mRNA during the embryonic development of Patella vulgata.

Abstract

As the first five cleavages of the Patella vulgata embryo are synchronous, they are well suited to determine the mRNA level of cyclin A and B genes in an embryo. During the third and fourth cleavage cycle the quantity of A and B mRNA is regulated in a cell-cycle-dependent way, reaching a high level between cleavages and a lower level just after mitosis. This implies that transcription of the cyclin genes occurs before the overall transcription increases directly after the fifth cleavage. During the first cleavages cyclin A and B mRNA is localized in distinct parts of the cytoplasm. Between two successive cell devisions it is found as a crescent-shaped domain at the peripheral side of the nucleus. At cytokinesis it is present between two separating nuclei and at newly formed cell membranes. At the fifth cleavage this localization disappears. Changes in the expression pattern of cyclin A and B may be expected after the fifth cleavage, when the first cells become arrested in cell division and differentiate. The mechanism causing cell division arrest of these primary trochoblasts is still unknown. Cell division arrest caused by the absence of cyclin A and/or B mRNA could be conditional for further differentiation. However, a decrease in cyclin A and B mRNA level in the trochoblasts is not detectable until 4 h after their last division. Later in development no cyclin A and B mRNA can be detected in these cells, whereas cyclin A and B mRNA is present in other cells of the embryo. Thus, the absence of cyclin A and B mRNA in primary trochoblasts, and in the later differentiating secondary and accessory trochoblasts is not obligatory for cell division arrest or cell differentiation.