Date of Award

2017

Document Type

Thesis

Degree Name

Bachelors

Department

Natural Sciences

First Advisor

Gilchrist, Sandra

Area of Concentration

Biology

Abstract

The impact of ocean acidification due to increased greenhouse gas emissions on ecosystems and the organisms which they comprise can be devastating. At the cellular level, pH changes can be dealt with in a number of ways, one of which is by the use of membrane bound proton pumps. This study examines the expression of a V-type proton ATPase in Ciona intestinalis in response to treatment with carbon dioxide. This species was chosen because it is widespread in marine habitats globally, and because it has been researched extensively for the effects of other marine pollutants. The V-type proton ATPase, which is expressed both in the endomembrane system and in the outer plasma membrane of cells, was tested for expression with primers for its catalytic A subunit. Organisms were cultured in a closed system tank of filtered sea water and fed a mixture of Phytoplex and Spirulina. Carbon dioxide gas was released in the tank such that the pH remained at 7.5 for the duration of the experiment. Three tunicates were collected, frozen using liquid nitrogen, and pulverized for each time point: 0 hrs treatment, 48 hrs, 72 hrs, 96 hrs, and 120 hrs. The RNA from each organism was then isolated, reverse transcribed, and analyzed using quantitative PCR targeting actin and the catalytic A subunit of the V-type ATPase. The results of PCR yielded only a few viable samples. However, expression of the ATPase was shown to increase over time with respect to the carbon dioxide treatment.

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