ABSTRACT
Cation-chloride cotransporters, including the Na-K-Cl cotransporter, play an important role in epithelial ion transport in insects. We have determined the tissue distribution of Manduca sexta bumetanide sensitive cotransporter (masBSC), a putative Na-K-Cl cotransporter that was originally cloned from M. sexta Malpighian tubules. We developed a polyclonal antibody (M6) against a C-terminal fragment of masBSC. masBSC protein was detected by M6 at an apparent molecular mass of approximately 220kDa in M. sexta foregut, midgut, hindgut, Malpighian tubule, salivary gland, fat body, trachea, and nerve cord. Higher expression was observed in the foregut than in other tissues. M6 stained the apical membrane of midgut epithelial cells in cross-sections of third instar larvae. The transcript of masBSC was detected by RT-PCR in midgut, Malpighian tubule, hindgut, trachea, nerve cord, and fat bodies. Taken together, these findings demonstrate that masBSC is widely expressed in M. sexta. While the specific function of masBSC remains unknown, the wide distribution indicates a role of masBSC in a broad range of tissues.
