Molecular investigations of BK(Ca) channels and the modulatory beta-subunits in porcine basilar and middle cerebral arteries

J Mol Histol. 2009 Apr;40(2):87-97. doi: 10.1007/s10735-009-9216-3. Epub 2009 Apr 1.

Abstract

Large conductance calcium-activated potassium (BK(Ca)) channels are fundamental in the regulation of cerebral vascular basal tone. We investigated the expression of the mRNA transcripts for the BK(Ca) channel and its modulatory beta-subunits (beta1-beta4) in porcine basilar and middle cerebral arteries using reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time PCR. Western blotting was used to detect immunoreactivity for the porcine BK(Ca) channel alpha-subunit and beta-subunit proteins. The BK(Ca) channel alpha-subunit RNA and protein distribution patterns were visualized using in situ hybridization and immunofluorescence studies, respectively. The study verified that the BK(Ca) channel alpha-subunit is located to smooth muscle cells of porcine basilar and middle cerebral arteries. The mRNA transcript for beta1-, beta2- and beta4-subunit were shown by RT-PCR in porcine basilar and middle cerebral arteries. However, at the protein level, only, the beta1-subunit protein was found by western blotting.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Basilar Artery / metabolism*
  • Blotting, Western
  • In Situ Hybridization
  • In Vitro Techniques
  • Large-Conductance Calcium-Activated Potassium Channel alpha Subunits / genetics
  • Large-Conductance Calcium-Activated Potassium Channel alpha Subunits / metabolism
  • Large-Conductance Calcium-Activated Potassium Channel beta Subunits / genetics
  • Large-Conductance Calcium-Activated Potassium Channel beta Subunits / metabolism
  • Large-Conductance Calcium-Activated Potassium Channels / genetics
  • Large-Conductance Calcium-Activated Potassium Channels / metabolism*
  • Middle Cerebral Artery / metabolism*
  • Oocytes / metabolism
  • Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Swine
  • Xenopus / metabolism

Substances

  • Large-Conductance Calcium-Activated Potassium Channel alpha Subunits
  • Large-Conductance Calcium-Activated Potassium Channel beta Subunits
  • Large-Conductance Calcium-Activated Potassium Channels