Biochemical Assessments of Cerebral Vasospasm: Measurement of cGMP, PKC, and PTK in Cerebral Arteries - Animal Models of Acute Neurological Injuries: Injury and Mechanistic Assessments - page 519

Fig. 1. Expression of PKC isoforms in native tissues. PKC isoforms expressed in rat brain (RB), canine brain (CB), and canine
basilar artery (BA) were examined by Western blotting. Samples were separated by SDS-PAGE (10% acrylamide gel) and
immunoblotted with each PKC isoform-specifi c antibody. To examine the antibody specifi city, antigen peptides were incu-
bated with corresponding primary antibodies, shown in the right of each blott (shown as “Peptide (+)”). Arrows indicate the
bands corresponding to each PKC isoform. Numbers on the left side of blot represent molecular weights (kDa). The follow-
ing PKC isoforms were detected; a , b 1, b 2, g , d , and e isoforms in rat brain, a , b 1, g , d , and e isoforms in canine brain, a
and d isoforms in canine basilar artery. From Nishizawa et al. ( 19 ) with permission.
PKC substrate. One way to examine the activity of specifi c PKC
isoform(s) is to measure the intracellular distribution as PKC trans-
locates from the cytosol to membrane fraction ( 23 ). Here, we
outline a method used to examine the intracellular distribution of
PKC isoforms using SDS-PAGE and Western blotting.
Recently, it has been reported that different stimuli activate
specifi c PKC isoform(s) causing translocation to distinct subcellular
compartments, which contributes to different physiological
phenomena ( 24-26 ). In the future, detailed fractionation of cell
membrane may provide precise information regarding the role of
PKC isoform(s) in cerebral vasospasm after SAH.