Sensory innervation of the cat knee articular capsule and cruciate ligament visualised using anterogradely transported wheat germ agglutinin–horseradish peroxidase

Abstract

Wheat germ agglutinin–horseradish peroxidase conjugate (WGA–HRP) was injected into the dorsal root ganglia (L5–S1) of the cat and used as an anterograde tracer substance for intra-axonal labelling of peripheral nerve endings in joint capsule and cranial (anterior) cruciate ligament (CCL). We believed that the high specificity of WGA–HRP for neural tissue along with the high visibility of its reaction product could help resolve controversies concerning the sensory innervation of the cruciate ligaments. Substantial amounts of WGA–HRP were transported in tibial nerve axons to the level of the knee. However, using standard HRP histochemistry we found that the capsular tissue and ligament synovia disintegrated during the incubation reaction. This problem was avoided by air drying the tissue slices on glass slides prior to reaction. Abundant labelling occurred in the posterior capsule with dense filling of axons and terminal endings. Sensory endings displayed features consistent with Ruffini endings and pacinian corpuscles. Sensory endings were located throughout the CCL in its sagittal plane, in the subsynovial layers and between collagen fascicles. In each CCL we observed 5–17 ovoid and elongated endings with dense terminal arborisations. These endings were between 100 and 150 μm long, were encapsulated, and gave rise to 1 or 2 axons. Large (up to 1.5 mm in maximum extent) elongated regions of dense, inhomogeneous labelling were found in the body of several CCLs. These resembled Golgi tendon-like endings, with the exception of their large size. We conclude that anterograde transport of HRP to the knee is a useful technique for labelling mechanoreceptors and axons in knee tissue. However, recently developed immunohistochemical analysis of peripheral tissue using protein gene product 9.5 appears to be the method of choice and should be employed for further study of human and animal cruciate ligament innervation.