The spatiotemporal development of innervation in spinal ligaments of chickens

The development of the innervation of both central and lateral (intertransverse) spinal ligaments was investigated in chickens between the time of hatching and 13 wk of age. A total of 36 White Leghorn chickens in 4 groups of 9 at ages 0, 2, 7, and 13 wk were used. The spinal ligaments were dissected, serially sectioned and labelled with a monoclonal antibody against neurofilament protein and observed using either conventional fluorescence or confocal microscopy. Only a few nerve elements were found in the ligaments at hatching and these consisted simply of single nerve fibres and small nerve bundles. The number of nerve elements increased rapidly up to 7 wk of age when large nerve bundles and Ruffini corpuscles were also found. The number of nerve elements decreased between 7 and 13 wk birds when the ligaments had begun to ossify and the amount of collagenous ligamentous tissue was significantly reduced. The fluctuation in numbers of neural elements was due to changes in numbers of single nerve fibres and small nerve bundles rather than large nerve bundles and Ruffini corpuscles which remained constant. In contrast to this significant increase in numbers of nerve elements, the innervation density of the ligaments decreased because of the overwhelming increase of the ligament volume due to growth. There were no differences between ligaments on the left and right sides of the body, but there was an unequal distribution of the neural elements within the ligaments; most were found in the cranial third of the intertransverse ligaments. These results show that significant changes in innervation of spinal ligaments occur during development and reflect the possibility that damage to the ligaments during this time could produce significant and permanent damage, especially in relation to the maintenance of an erect spine.