Continued lateral mobility of the flanking paranodal domains ultimately results in axonal domain disorganization, including disrupted paranodal axo-glial junctions, as well as pinching of the nodal axolemma. Thus, NF186 acts to delineate the nodal region by coordinating the organization and assembly of several transmembrane Dolutegravir order and cytoskeletal proteins into a unique molecular

domain, which is then further stabilized by interactions with the glial processes overlying the nodal region. These mechanisms point to an independent, and not interdependent, assembly of the nodal complex in vivo, which, once established, serves as a molecular fence to maintain distinct boundaries to facilitate saltatory conduction along myelinated fibers. All procedures involving mice were carried out under UNC-IACUC approved guidelines for the ethical treatment of laboratory animals. The Nefl-Cre mice were generously provided by Dr. Michael Sendtner (University of Cilengitide chemical structure Wurzburg, Germany) and were previously described ( Schweizer et al., 2002). The Cnp-Cre;NfascFlox and the Caspr −/− mice were previously

described ( Pillai et al., 2009 and Bhat et al., 2001). The TaumGFP/LacZ mice ( Hippenmeyer et al., 2005) were provided by Dr. William Snider (University of North Carolina). The Rosa26RLacZ (R26RLacZ) mice were generously provided by Dr. Victoria Bautch (University of North Carolina). The β-Actin-Cre (Act-Cre) mice were obtained from Jackson Labs (ME). The NfascFlox mice

used in this study were MTMR9 previously generated and described ( Pillai et al., 2009). For extraction of genomic DNA from tail and spinal cord samples, the REDExtract-N-Amp Tissue PCR kit was used according to the manufacturer’s directions (Sigma-Aldrich, USA). Primers used include: Nfasc primer forward primer 1,5′-TTTCTGACTGTTCTGGGTGAC-3′ and reverse primer 2,5′-GCTACGATGTATCATTTGGCAG-3′; the Null forward primer 3,5′-TTTACGGTATCGCCGCTCCCGATT-3′; and the Null reverse primer 4,5′-CCCTGTTCTGCTCCTGGTTCAGTC-3′. For Cre, we used the primers previously described ( Pillai et al., 2009). The following antisera were previously described: guinea pig and rabbit anti-Caspr (Bhat et al., 2001), guinea pig anti-NF186 (recognizing the mucin domain), and rat anti-pan Neurofascin (NFct, recognizing the C terminus) (Pillai et al., 2009); rabbit anti-Caspr2; and rat anti- AnkG (Thaxton et al., 2010). Additional primary antibodies used include mouse anti-pan-Nav), mouse anti-Dystrophin (Dp116, MANDRA1), mouse anti-Act, and mouse anti-potassium channel (Kv1.1) from Sigma; and rabbit anti-NrCAM, rabbit anti-EBP50, rabbit anti-Gldn, and mouse anti-myelin basic protein (MBP) from Abcam. Mouse anti-β-Tubulin (Tub) was obtained from Cell Signaling. Rabbit anti-FIGQY was generously provided by Dr. Matt Rasband (Baylor College of Medicine).