Immunolocalization of Ca2+ -activated potassium channel beta4 subunit in neuronal mitochondria

Marta E. Głąb ,  Grzegorz M. Wilczyński ,  Adam M. Szewczyk 

Instytut Biologii Doświadczalnej PAN (IBD), Pasteura 3, Warszawa 02-093, Poland

Abstract

Large conductance Ca2+-activated potassium channels (BKCa channels) are widely expressed in plasma membrane of various cell types. Interestingly, recent studies provided the evidence for the existence of BKCa channels also in mitochondria. However, the molecular composition of these channels as well as their cellular and tissue distribution is still unknown. Hence, the goal of the present study was to search for the potential candidate for the regulatory component of neuronal mitoBKCa channel. A combined approach of Western Blot analysis, immunofluorescence and electron microscopy with the use of antibodies directed against four distinct betasubunits demonstrated the presence of BKCa channel beta4 subunit (KCNMB4) in the inner membrane of neuronal mitochondria. Double-labeling immunocytochemistry in paraffin embedded rat brain sections as well as in primary culture of rat hippocampal dentate gyrus revealed the colocalization of BKCa channel beta4 subunit with mitochondrial staining. Furthermore, the distribution pattern of beta4 subunit indicated that its expression is restricted to the subpopulation of mitochondria within the cell. Electron microscopy data additionally confirmed mitochondrial residence of beta4 and proved its close association with inner mitochondrial membrane. These results therefore suggest that beta4 subunit could be a potential candidate for the regulatory component of mitochondrial BKCa channels in neurons.

This study was supported by the Ministry of Science and Higher Education grant No. PBZ-MIN-001/P05/11
 

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Presentation: Poster at Zjazd Polskiego Towarzystwa Biochemicznego, Sympozjum H, by Marta E. Głąb
See On-line Journal of Zjazd Polskiego Towarzystwa Biochemicznego

Submitted: 2007-04-26 18:20
Revised:   2009-06-07 00:44