Meeting Abstract
P1.129 Thursday, Jan. 3 Microarray analysis of hepatopancreas heat stress responses in thermally acclimated porcelain crabs, Petrolisthes cinctipes TOMAS MIRANDA, Claudia P.*; GALASSI, Eric W.; STILLMAN, Jonathon H.; San Francisco State University; San Francisco State University; San Francisco State Unversity claudt85@hotmail.com
Intertidal zone organisms experience thermal stress during periods of low tide, and much work has shown that induction of heat shock proteins occurs in response to this stress. Here, we investigated the plasticity of gene expression responses to heat shock in porcelain crabs that were acclimated to 8°, 14°, and 22° C. We used cDNA microarrays containing a library of Petrolisthes cinctipes ESTs representing 6,400 unique consensus sequences to identify genes that are differentially expressed during recovery in heat stressed crabs relative to crabs held at their acclimation temperature. Crabs from each acclimation temperature were given a heat stress or held at their acclimation temperature for 4h, and then were placed back into their acclimation aquarium for recovery. Individuals were sampled at various recovery timepoints. Sampling consisted of dissecting tissues into liquid nitrogen. Here, RNA was extracted from the hepatopancreas tissue using TRIZOL, 10�g were reverse transcribed into amino allyl cDNA, labeled with Cy3 and Cy5 dyes, and hybridized to cDNA microarrays. Each hybridization contained a heat stress sample and a control sample from the same acclimation temperature. Due to limited samples at the 1h recovery timepoint we used pooled RNAs for comparisons of heat stress responses across acclimation temperatures. We also compared using individual vs. pooled samples at one acclimation temperature to see how pooling affected identification of differentially expressed genes. Statistical identification of differentially expressed genes were determined with R/MAANOVA, and the results of these analyses presented here. This work was funded by NSF 0533920 to JHS.
