TERANISHI, Kristen S*; STILLMAN, Jonathon H; University of Hawaii at Manoa; Romberg Tiburon Center, San Francisco State University: Seasonal variation in hepatopancreas transcriptome profiles following heat stress in the porcelain crab, Petrolisthes cinctipes

How organisms respond to thermal stress in their natural habitat is a function of thermal acclimatization state. Here, we have investigated seasonal differences in transcriptome responses to acute thermal stress in one population of the intertidal porcelain crab, Petrolisthes cinctipes. In winter and summer 2005, crabs were collected from the field site at Cape Arago, Oregon where n=6 crabs were immediately dissected and additional crabs were taken to the laboratory and divided into 2 groups. One group was subjected to a 4h thermal ramp to 30�C, and the other group was held at a constant temperature (10-12�C) during the 4h treatment. Following the 4h treatment, crabs were placed into a recovery tank (10-12�C) and n=5 crabs from each group were sampled after 6h. For each crab sampled, hepatopancreas tissue was dissected immediately into RNA tissue protect (Qiagen), RNA was purified using Qiagen RNeasy spin columns, and reverse transcription was used to synthesize amino-allyl cDNA. Amino allyl cDNAs were labeled with Cy3 or Cy5 dye and hybridized on microarrays using a dye-swap design for each season. The microarray used for this study was printed from a 13,824-clone cDNA library created from a range of P. cinctipes tissue types (including whole crabs, and tissues from gill, heart, muscle, nerve, and hepatopancreas) from crabs exposed to a range of thermal conditions. Fluorescence data were log transformed and normalized using the Loess function. MAANOVA in R was employed to identify genes that had significantly different expression profiles between field acclimatized, thermally stressed and control crabs between seasons. This research was supported by a NSF Graduate Research Fellowship to KST and NSF IOB-0533920 and JGI CSP to JHS.