The presence of intact paranodal junctions on myelinated axons in the CNS and PNS is crucial for both myelin sheath attachment and saltatory impulse conduction. The axonal glycoprotein contactin-associated protein (Caspr) is expressed in the paranodal region and plays an important role in the creation and maintenance of these adhesive junctions. In the present study, antibodies to Caspr were used to assess the integrity of paranodal junctions on myelinated axons in brain and spinal cord tissue from subjects with longstanding multiple sclerosis, a neurological disorder that affects both myelin and axons. Triple immunofluorescence combined with confocal laser scanning microscopy showed that axons in the demyelinated centre of the 36 brain and 16 spinal cord multiple sclerosis lesions studied were devoid of Caspr immunoreactivity, suggesting that axons down regulate the expression of Caspr following demyelination. Additional data indicated that Caspr reappears in the paranodal region with the formation of new myelin sheaths. Immuno labelling further revealed that Caspr on myelinated axons in border regions was often no longer concentrated in the paranodal region, but was also present in the internodal region-a phenomenon particularly common in the borders of the more chronic lesions in the collection. Myelinated axons with long Caspr-positive stretches were often present at a considerable distance from the lesion edges. These findings raise the possibility that the aberrant location of Caspr is an early sign of impending myelin loss. This would imply that demyelination continues at a slow rate in established lesions. The diameters of Caspr-positive structures on some myelinated axons near the lesion edges were also increased. Moreover, the gap between individual myelin sheaths on these apparently swollen axons was widened occasionally and a very small myelin sheath plus additional Caspr-positive structures had sometimes formed in the enlarged space. This finding thus suggests that the formation of new myelin in multiple sclerosis is not only induced following the loss of complete internodes but also in response to broadening of the nodal region. Interestingly, alterations in the expression and localization of Caspr were observed in tissue from both subjects with the primary and secondary progressive form of multiple sclerosis. In summary, the present study provides immunohistochemical evidence that paranodal junctions on some myelinated axons in the borders of lesions of patients with chronic progressive multiple sclerosis are no longer intact. This may impair saltatory impulse conduction and lead to further myelin loss, thereby contributing to disease progression in multiple sclerosis.