Journal: Biomedicines
Article Title: Compromised Myelin and Axonal Molecular Organization Following Adult-Onset Sulfatide Depletion
doi: 10.3390/biomedicines11051431
Figure Lengend Snippet: Mislocalization of axonal domain proteins following adult-onset sulfatide depletion. ( A ) An axon from a CTL mouse demonstrates proper ion channel and axonal domain protein organization in the spinal cord with Nav1.6 (green) localized to the node of Ranvier, Caspr1 (blue) localized to the paranode and Kv1.1 (red) localized to the juxtaparanode. Common, but not exclusive, pathologies observed included ( B ) Kv1.1 invading the paranodal region (white arrows); ( C ) elongated sodium channel domain (white star) and heminode (white bracket) (note: z-stack collected encompassed entirety of fluorescence and absent paranode was not the result of exclusion of z-stack); ( D ) elongated Caspr1 localization (white brackets); ( E ) combination of several pathologies including binodal sodium channels clusters (yellow stars) and intermixing of Kv1.1 channels and Caspr1 (white star). ( F ) Quantification of protein organization revealed preservation of protein domains at 3 months PI but a significant loss of axonal domain organization at 6 and 11 months PI. Data are presented as mean ± standard error; ** p < 0.01 **** p < 0.0001. Scale bars = 5 µm.
Article Snippet: Cervical spinal cords were cryopreserved in 30% sucrose in 1× phosphate buffered saline (PBS), frozen in Tissue Tek Optimal Cutting Temperature media (Sakura, Torrance, CA, USA) [ ], sectioned at 40 µm and triple immunolabeled with antibodies directed against voltage-gated sodium channel type 1.6 (Nav1.6; 1:250, mouse monoclonal IgG1, Antibodies Inc., Davis CA, USA; cat# 75-026), Caspr1 (1:500, rabbit polyclonal, Abcam cat# ab34151) and voltage-gated potassium channel type 1.1 (Kv1.1; 1:750; mouse monoclonal IgG2b, Antibodies Inc., cat# 75-105).
Techniques: Fluorescence, Preserving
