Patterning the Mud Snail Ilyanassa obsoleta: The Role of Cell Signaling and Asymmetric Protein Localization

Abstract

The polar lobe of Ilyanassa is asymmetrically partitioned into the D lineage of cells. Two of these cells, 3D and 4d, induce proper axial cell fate patterning in the embryo. Based on known embryological data in Ilyanassa, I hypothesized that Notch signaling would be required for this induction. I found that Notch signaling is required for cell fates induced by 4d and is temporally required well after 4d induction. Based on these results, I hypothesize that Notch signaling is involved in a reciprocal induction between the micromeres and the macromeres (endoderm) resulting in the maintenance of micromere fate induction and endoderm specification.Loss of the polar lobe results in the loss of cell fate induction by 3D/4d. Therefore, I hypothesized that proteins are asymmetrically bequeathed to the inducing D lineage cells by the polar lobe. To test this hypothesis, I compared global protein differences between two cell stage intact embryos, lobeless embryos, and isolated polar lobes by 2-Dimensional Electrophoresis analysis. I found several (12) quantitative differences between these samples including four spots enriched in the polar lobe isolates. I identified voltage-dependent anion-selective channel (VDAC) as one of the candidate proteins enriched in polar lobe isolates. I propose that VDAC is asymmetrically distributed by the polar lobe to the D cell and that it may function in D cell induction and mesendoderm fate specification.Lastly, I identify an acetylated tubulin antigen as a marker for cilia. I describe the pattern of cilia differentiation in the developing larvae that results in the formation of two ciliary bands, the prototroch and the metatroch, required for locomotion and feeding. These ciliary bands are conserved among annelid and mollusc larvae. Interestingly, the metatroch is derived from third quartet derivatives in the annelid Polygordius and from second quartet derivatives in the mollusc Crepidula. I provide evidence that the metatroch is derived from the first quartet derivatives in the mollusc Ilyanassa. Thus while the larval metatroch is conserved, its clonal origin is not. Based on these results, I provide support for the hypothesis that the metatroch is not homologous between annelids and molluscs or even among molluscs.