Differential expression of eight transcripts and their roles in the cuticle of the blue crab, Callinectes sapidus

Eight cuticle protein transcripts from Callinectes sapidus were sequenced and their expression determined across the molt cycle in both calcifying and arthrodial cuticle hypodermis using quantitative PCR, Northern blots, and in situ hybridization. Four transcripts, designated CsAMP, are found only in non-calcifying arthrodial membrane hypodermis. They all code for a Rebers-Riddiford-1 motif, known to bind chitin. CsAMP9.3 is most likely an exocuticle constituent since it is expressed only during pre-molt. The other three arthrodial transcripts are present both before and after ecdysis. One of these, CsAMP16.3, codes for a RGD cell-attachment motif that could be involved in anchoring chitin-protein fibers to pore canals, cellular extensions of the hypodermis in the cuticle. The other four transcripts, designated CsCP, were found only in calcifying hypodermis. CsCP14.1 contains an RR-1 motif, which is more commonly found in non-calcifying cuticle proteins. CsCP6.1 is expressed post-molt and contains a partial RR motif, suggesting that it could bind to chitin in the endocuticle. The other two transcripts from calcifying hypodermis do not code for RR proteins, but both contain three copies of a different insect cuticle motif. One of these, CsCP19.0, is expressed only post-molt while the other, CsCP15.0, is present both before and after ecdysis.