Bioelectricity refers to differential membrane voltage and cytoplasmic ion concentrations in tissues or cells which persist over long periods of time. Differences in these steady-state ionic conditions are responsible for large-scale axial patterning and morphogenesis in developing embryos. The sea urchin embryo is an excellent model organism for studying embryonic development, yet a comprehensive study of bioelectricity in sea urchin development has not been reported. Differential ion channel activity is a primary means by which bioelectricity is controlled; thus, we hypothesized that disrupting ion channel activity would reveal the requirements for bioelectricity in the sea urchin embryo. We performed a screen of ion channel inhibitors and discovered that their activities are required for many processes in sea urchin development. We chose two interesting phenotypes to investigate further. First, we demonstrate that H+/K+ ATPase (HKA) activity is required for biomineralization of the sea urchin larval skeleton. We determined that embryos raised with HKA inhibitors initially exhibit voltage and pH changes, then revert to normal voltage and pH during biomineralization via compensatory changes in sodium and chloride ions; it is likely that these compensatory changes lead to defects in transport of carbonate ions, that in turn, inhibit biomineralization of the calcium carbonate skeleton. We hypothesize that similar mechanisms are at play in human patients on long-term HKA inhibitors to treat acid reflux, in whom biomineralization is also decreased. Next, we demonstrate that V-type H+ ATPase (VHA) activity is required for specification of the dorsal-ventral (DV) axis, for the normal inactivation of p38 MAPK in the presumptive dorsal region, and for the subsequent asymmetric onset of expression of the TGFβ family member Nodal, that locally specifies the ventral territory. Embryos treated with VHA inhibitors exhibit global p38 MAPK activity and Nodal expression, and are ventralized. We describe previously unknown gradients of voltage and pH across the DV axis, the sharpness of which requires VHA activity. We propose that the voltage and pH gradients encode spatial information which confers asymmetry on p38 MAPK activity. Overall, we demonstrate that bioelectrical changes are essential for development of the sea urchin embryo, specifically via roles in biomineralization and DV axis specification. / 2019-01-25
| Identifer | oai:union.ndltd.org:bu.edu/oai:open.bu.edu:2144/27332 |
| Date | 26 January 2018 |
| Creators | Schatzberg, Daphne |
| Source Sets | Boston University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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