The Expression Profiles of Zebra Mussel Byssus Genes along Byssogenesis

To identify the expression profiles of zebra mussel byssus unique genes with the regeneration of the byssal threads, a time-course study was performed. The RNA samples from the feet of fully attached zebra mussels (maintained attachment for more than 4 weeks) were extracted as common reference. The detached mussels were allowed to re-attach on underwater surface. Then the RNA samples from feet of the mussels at different time points post detachment were taken, namely, 12 hours, 1 day, 2 days, 3 days, 4 days, 7 days, and 21 days post detachment. The comparisons of the genes expression profiles were made between different time points and reference, as well as the any two connected time points (expect for 7 days and 21 days). The dual-channel microarray hybridizations were performed as following: 12h→R, 12h←R, 1D→R, 1D←R, 2D→R, 2D←R, 3D→R, 3D←R, 4D→R 4D←R, 7D→R, 7D←R, 21D→R, 21D←R, 12h→1D, 12h←1D, 1D→2D, 1D←2D, 2D→3D, 2D←3D, 3D→4D, 3D←4D, 4D→7D, 4D←7D, 7D→21D, 7D←21D. The direction of the arrows stand for the way of labeling: Alexa555→Alexa647. Six biological replicates of 1D, 2D, 3D, 4D, and 7D were used; four biological repllicates of 12h and 21D were taken; fourteen biological replicates of reference were applied. The whole experiment includes 26 microarray slide hybridizations.