Available at: http://digitalcommons.calpoly.edu/theses/443
Date of Award
MS in Biological Sciences
The sea squirts Ciona intestinalis and C. savignyi have disparate distribution patterns, which may result from differences in their thermal tolerance limits. Because C. intestinalis, an almost cosmopolitan species, has a more widespread distribution, it is thought that it is better adapted to endure a wide range of temperatures. In order to compare the heat stress response between these two congeners, we studied global changes in protein expression, using a proteomics approach. To characterize the response to extreme heat stress, animals of both species were exposed to temperatures of 22°C, 25°C, and 28°C for 6 h, and then were left to recover at a control temperature (13°C) for 16 h. An additional experiment was conducted to assess the effect of mild-to-moderate heat stress including a 6 h exposure to temperatures of 18°C, 20°C, and 23°C, and a 16 h recovery at a control temperature (16°C). A quantitative analysis, using 2D gel electrophoresis and gel-image analysis, showed that in the high heat stress (HHS) experiment, 15% and 18% of the all protein spots detected demonstrated changes in expression in C. intestinalis and C. savignyi, respectively. In the low heat stress (LHS) experiment, 4% of the total number of proteins detected changed significantly in both C. intestinalis and C. savignyi. Using matrix-assisted laser desorption ionization (MALDI) tandem time-of-flight mass spectrometry, we were able to identify proteins with a 65-100% success rate, depending on species. Our results indicate that C. intestinalis maintains higher baseline levels of molecular chaperones and launches a quicker response to thermal stress than C. savignyi, suggesting it may be the more thermally tolerant of the two. In addition, actins, tubulins, and ATP-synthase F1 β-subunits were the most susceptible to proteolytic degradation, which may indicate that they have relatively higher thermal sensitivities.