Electron microscopical investigations of the intercellular contacts during the early cleavage stages ofLymnaea stagnalis (Mollusca, Gastropoda).

In early cleavage stages ofLymnaea stagnalis, three kinds of intercellular junctions could be distinguished up to the sixth cleavage: intermediate, septate and gap junctions. The first two form "junctional belts" located on the cell border at the periphery of the embryo. For the purpose of our study we were most interested in gap junctions as they are alleged to be structures that allow cell-to-cell communication. Gap junctions first appear at the four cell stage. Up to the sixth cleavage no difference in the distribution pattern could be found between and within each of the four quadrants of the embryo. Some of the cell tiers along the animal-vegetal axis lack gap junctions either between the blastomeres within the tier or between the blastomeres from adjacent tiers. All gap junctions observed in freeze fracture replicas show plaques with an irregular IMP pattern. The average IMP diameter measures 12 nm (SD±2 nm). In stages fixed after the fifth cleavage, gap junctions are found between micromeres at the animal pole and the central 3D macromere. This is in agreement with the presumed interaction between these cells at this stage. The possibility of a transition of non-functional into functional gap junctions after the fifth cleavage is discussed.

Communication compartments in the ectoderm of embryos of Patella vulgata.

Patterns of gap junctional communication in the ectoderm of embryos of Patella vulgata have been studied by intracellular injection of the fluorescent dye Lucifer Yellow, and by analysis of its subsequent spread to adjacent cells (dye-coupling). We found that dye-coupling became progressively restricted to different domains of the ectoderm, forming communication compartments. These communication compartments are characterized by their high coupling abilities within the compartment, and reduction of coupling across their boundaries. During development, the pretrochal (anterior) ectoderm becomes subdivided into two communication compartments, the apical organ and the anlage of the head ectoderm. The posttrochal (posterior) ectoderm becomes subdivided into different communication compartments in two successive phases. Firstly, in the 15-h embryo the dorsal and ventral domains of the ectoderm form separate communication compartments. A dorso-ventral communication boundary restricts the passage of dye between the two domains. Secondly, in the 24-h embryo dye-coupling becomes further compartmentalized in both the dorsal and ventral domains. These compartments correspond to the anlagen of different ectodermal structures. In order to study whether any level of coupling persists between the ectodermal compartments we injected currents through a microelectrode inserted into one cell of one compartment and monitored its spread by means of a second microelectrode inserted into one cell of another compartment (electrical coupling). Despite the absence of dye-coupling, electrical coupling between the ectodermal dye-coupling compartments was detected, which suggests that some level of communication is maintained between compartments. Our results demonstrate that within the ectoderm layer of Patella vulgata the transfer of dyes becomes progressively restricted to communication compartments and, concomitantly with the specification of the different ectodermal anlagen, these compartments become subdivided into smaller communication compartments.