Date of Award

2021

Degree Type

Dissertation

Degree Name

Doctor of Philosophy in Biological and Environmental Sciences

Department

Cell & Molecular Biology

First Advisor

Steven Irvine

Abstract

This dissertation is comprised of two parts, the first is cell culture. In this work I describe the successful primary culture of Ciona intestinalis ovary tissue. Improved cell proliferation in 3D Petri Dish® system over culture treated plastic is shown, as well as the efficacy of using C. intestinalis hemolymph as a media additive. The seeded ovarian cells assemble into multicellular aggregations, which mimic the morphology of in vivo ovaries. It is also important to understand the molecular pathways in which marine organisms handle stress and how these pathways change in high stress situations.

The second is the role of Shp2 in temperature stress. It is projected that ocean temperatures may raise as much as 4C over the next century, and there is very little known about how this increase in temperature will affect both reproduction and embryonic development in marine organisms. In this work I also describe the phenotypic abnormalities which occur during embryogenesis in Ciona intestinalis when Shp2 protein tyrosine phosphatase is inhibited. The phenotypic abnormalities are consistent with the abnormalities observed in C. intestinalis embryos reared at high temperatures stress. This suggests that Shp2 is an important protein in the amelioration of stress in developing C. intestinalis embryogenesis and is required for normal embryogenesis.

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