Repository for transcriptomic work related to 2021 Pycnopodia coelomocyte RNAseq.
In the summer of 2021, adult Pycnopodia helianthoides were collected (Permit: Harvell 21-1172) by a team of WDFW divers and brought to USGS Marrowstone Field Station.
Stars were observed for several weeks and any that showed signs of disease were not used for experiments.
Several experiments were performed, but I have RNAseq data from Experiment 2 - details below.
Questions:
- Is Sea Star Wasting Disease transmissible?
- If yes, is the causative agent in the small size fraction?
Multiple tissue types from an adult Pycnopodia helianthoides wasting from the field were blended together using a Tissue Tearor (link; Tissue Tearor 985370-07 Homogenizer; 120 VAC, 1.2 A). The sample was then spun down to pellet larger bits of tissue. The supernatant was then used as the inoculum for experiments.
The following inoculate types were made:
- Heat-killed (control): supernatant was placed in a falcon tube and held in boiling water with a stir bar for 10 minutes
- 0.45um filtered (exposed): supernatant was filtered through a vacuum bottle unit - flow-through is larger viruses and bacteria
- unfiltered (exposed): supernatant was unfiltered, so it would contain anything from a large protist to a small virus
Due to this summer being in the very early stages of our group's work, aspects of this work was a bit more on the exploratory side and there were several different inoculation dates.
- Control: n=13 stars injected with heat-killed inoculum
- Exposed: n=12 stars injected with 0.45um filtered live inoculum
- Exposed: n=4 stars injected with unfiltered live inoculum
- Control: n=9 stars injected with heat-killed inoculum
- Exposed: n=9 stars injected with unfiltered live inoculum