Wulf Helle, Hay-Schmidt Anders, Poulsen Asser Nyander, Klaerke Dan Arne, Olesen Jes, Jansen-Olesen Inger
Department of Neurology & Danish Headache Center, Glostrup Research Institute, Glostrup Hospital, Faculty of Health Sciences, University of Copenhagen, 2600 Glostrup, Denmark.
J Mol Histol. 2009 Apr;40(2):87-97. doi: 10.1007/s10735-009-9216-3. Epub 2009 Apr 1.
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.
大电导钙激活钾（BK(Ca)）通道在脑血管基础张力的调节中起着基础性作用。我们使用逆转录聚合酶链反应（RT-PCR）和定量实时PCR，研究了猪基底动脉和大脑中动脉中BK(Ca)通道及其调节性β亚基（β1-β4）的mRNA转录本表达情况。采用蛋白质免疫印迹法检测猪BK(Ca)通道α亚基和β亚基蛋白的免疫反应性。分别使用原位杂交和免疫荧光研究观察BK(Ca)通道α亚基RNA和蛋白质的分布模式。该研究证实，BK(Ca)通道α亚基定位于猪基底动脉和大脑中动脉的平滑肌细胞。RT-PCR显示猪基底动脉和大脑中动脉中有β1、β2和β4亚基的mRNA转录本。然而，在蛋白质水平上，蛋白质免疫印迹法仅检测到β1亚基蛋白。
