Freshly isolated bovine coronary endothelial cells do not express the BKca channel gene

Recent reports have suggested that different types of Ca(2+)-activated K(+) channels may be selectively expressed either in the vascular endothelial cells (ECs) or smooth muscle cells (SMCs) of a single artery. In this study, we directly compared mRNA, protein and functional expression of the high-conductance Ca(2+)-activated K(+) (BK(Ca)) channel between freshly isolated ECs and SMCs from bovine coronary arteries. Fresh ECs and SMCs were enzymatically isolated, and their separation verified by immunofluorescent detection of alpha-actin and platelet/endothelium cell adhesion molecule (PECAM) proteins, respectively. Subsequently, studies using a sequence-specific antibody directed against the pore-forming alpha-subunit of the BK(Ca) channel only detected its expression in the SMCs, whereas PECAM-positive ECs were devoid of the alpha-subunit protein. Additionally, multicell RT-PCR performed using cDNA derived from either SMCs or ECs only detected mRNA encoding the BK(Ca) alpha-subunit in the SMCs. Finally, whole-cell recordings of outward K(+) current detected a prominent iberiotoxin-sensitive BK(Ca) current in SMCs that was absent in ECs, and the BK(Ca) channel opener NS 1619 only enhanced K(+) current in the SMCs. Thus, bovine coronary SMCs densely express BK(Ca) channels whereas adjacent ECs in the same artery appear to lack the expression of the BK(Ca) channel gene. These findings indicate a cell-specific distribution of Ca(2+)-activated K(+) channels in SMCs and ECs from a single arterial site.