Figure 4.

Detection of different brevican fragments. (a) Immunoreaction with pan-specific brevican antibodies (BD Bioscience, FL) clearly surrounds DCN neurons in wt but not in pcd mice. As internal control nontarget region of PC axons, the cochlear nucleus shows no alterations in immunoreactions with FL. (b) The 50 kDa isoform of brevican seems to be nearly absent around DCN neurons of pcd mice, whereas the not affected region (CN) revealed brevican-bearing neurons. (c) The B756 antibodies detect mainly the 80/90 kDa and full length isoforms of brevican. These cleavage products aggregate around the neurons in DCN of wt mice and seem to be integrated in PNs. In pcd, PNs appear with lower intensity, but with potential higher parenchymatic reaction. PN-detection with all three antibodies in the internal nontarget control region (CN) is unchanged. Scale bar: 20 μm. (d) Biochemical detection of brevican with SDS-PAGE with pan-specific antibodies revealed most known isoforms at 50, 80, 90, and 145 kDa. Quantification of the 50 and 145 kDa brevican isoform showed a significant increased protein expression in pcd (p < 0.001), respectively, for the different isoforms. Therefore, the diagram is supposed to display the optical density (OD) values of pan-brevican chemiluminescent signal summed values (OD 50 kDa + 145 kDa brevican/actin). Data are given as mean ± SEM.