title: Identification and quantification of full-length BK channel variants in the developing mouse cochlea.
abstract: Maxi-K(+) (BK) channel diversity is attributed to alternative splicing in the kcnma1 gene. The resultant variants manifest themselves in different cell types, tissues, and functions, such as excitation, metabolism, and signaling. Immunoelectron microscopy revealed immunogold particle labeling of BK in apical and basal regions of inner and outer hair cells, respectively. Additional labeling occurs in Deiters' cells and the inner mitochondrial membrane. Identification of full-length sequences reveals 27 BK variants from embryonic and postnatal mouse inner ear, per classification by tail motif, VYR, DEC, and ERL, and by exon usage. Three predicted start codons are found encoding MAN, MSS, and MDA, of which MDA shows the greatest expression through all stages in development, whereas MAN is undetectable. Complex splice sites occur between exons 9 and 10 and between 21 and 23. Spliced-in/out exons between 8 and 10 reveal a short fragment composed of exons 8 + 10, detectable on postnatal day (PD) 14 and PD30, and a longer fragment composed of exons 8 + 9 + 10 that is upregulated on embryonic day (ED) 14. Spliced-in exons 22 or 23 are expressed on ED14 but decrease over time; however, exon 22 increases again on PD34. Using tail-specific primers, qRT-PCR from ED14, PD4, -14, and -30 shows that BK-VYR and -ERL dominate expression on ED14, whereas DEC dominates after birth in all cochlear regions. The localization of BK and the changes in expression of its exons and tail types, by alternative splicing during development, may contribute to cochlear organization, acquisition of hearing, and intracellular signaling.
authors.affiliation: Department of Otolaryngology-Head and Neck Surgery, University of South Florida, Tampa, Florida 33612, USA.
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Sakai
authors.forename: Margaret
authors.initials: M
authors.lastname: Harvey
authors.forename: Bernd
authors.initials: B
authors.lastname: Sokolowski
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D019976:Cloning, Organism; D003051:Cochlea / Q000196:embryology / Q000254:growth & development / Q000378:metabolism*; D005091:Exons; D005260:Female; D051036:Large-Conductance Calcium-Activated Potassium Channels / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism*
chemical_list: D051036:Large-Conductance Calcium-Activated Potassium Channels; D020033:Protein Isoforms