Dorsal-Ventral Patterning in the Mud Snail, Ilyanassa obsoleta

The experiments reported here describe mechanisms involved in the establishment of the dorsal-ventral axis in the mud snail, Ilyanassa obsoleta. Ilyanassa and other spiralians utilize an embryonic organizer to induce dorsal identity, and thus establish the bilateral axis. The D macromere embryonic organizer in Ilyanassa is specified at the four-cell stage by the inheritance of the polar lobe, but does not function as an inductive center until the 24-cell stage. Previously it was assumed that the D macromere of Ilyanassa functioned autonomously through its inheritance of the polar lobe. I have found this is not the case. Rather, I describe the role that the micromeres play in the activation of the D macromere organizer. Specifically, I have found that micromeres of the first and second quartet are necessary for at least three known characteristics of the D macromere: the activation of MAPK in the D macromere, the division of the D macromere, and the inductive capacity of the D macromere. Thus, while the polar lobe is necessary for D macromere function, its inheritance does not provide the D macromere with functional autonomy.The localized activation of MAPK was the first molecular component of dorsal-ventral patterning to be identified in Ilyanassa and other spiralians. In addition to being activated in the D macromere organizer, MAPK is also activated in the micromeres that are induced by the D macromere. I undertook a pharmacological screen to identify other components involved in dorsal-ventral patterning. I have found that a member of the Protein Kinase C (PKC) family is also involved in the establishment of the dorsal-ventral axis in Ilyanassa. Inhibition of PKC disrupts patterning, resulting in a radialized animal. In addition, I have found that PKC functions in the same path as MAPK. PKC is necessary for the proper activation of MAPK in the D macromere organizer and the micromeres. These results suggest that either the same transduction pathway is used repeatedly in the establishment of the dorsal-ventral axis or that patterning is the result of one global signal. These results drastically change our view of dorsal-ventral patterning during spiralian development