BK-type calcium-activated potassium channels are large conductance channels that respond to changes in intracellular calcium and membrane potential. These channels are used in a wide variety of cell types and have recently been linked to drug sensitivity and tolerance. In both Drosophila and mammals, BK channels are encoded by the slowpoke gene. The Drosophila slowpoke gene includes 14 alternative exons distributed among five sites of alternative splicing. Presumably, the purpose of alternative processing is to provide transcripts tailored to the needs of the cell. The slowpoke gene is expressed in nervous, muscle and epithelial tissues. To determine whether splicing is controlled in a tissue- and/or developmental-specific manner, we built tissue- and developmental-specific cDNA libraries that preserved the relative frequency of various slowpoke splice variants. These libraries were screened by colony hybridization using alternative exon-specific DNA probes to document the frequency of individual alternative exons in different developmental stages and distinct tissue types. We demonstrate that slowpoke transcripts undergo tissue- and developmental-specific splicing in Drosophila and some exons are diagnostic for specific tissues.