RESUMO
OBJECTIVE: To evaluate the morphology, distribution and function of palisade endings (PE) in human extraocular muscles (EOM), and observe the alterations in eyes with congenital nystagmus (CN). The etiology and pathogenesis of CN were also investigated. METHODS: It was a experimental study. The distal myotendinous junctions of the EOM were obtained during operation for CN (CN group) and concomitant strabismus (control group). The samples from patients with similar age and same extraction sites in the two groups were compared. The muscles cut during operation were immediately put into 4% glutaraldehyde fixative solution. And 1-2 transverse bands of tissue were cut every 1 mm from tendon insertion for specimens processing. The ultrastructure of EOM and PE in the two groups was observed by transmission electron microscopy. The distal parts of EOM cut during operation were put into 4% paraformaldehyde promptly. Myotendinous junction region whole mounts were labeled with antibodies against choline acetyltransferase (ChAT). Muscle fibers were counterstained with phalloidin. And longitudinal and transverse cryostat serial sections were cut at 25 µm and analyzed by confocal laser scanning microscopy. The ChAT expression, morphology and distribution of PE were observed. The same fragment of myotendinous junction in the two groups was selected. After the total protein was extracted, ChAT was detected by western blot. The expression level of ChAT was analyzed. RESULTS: Compared with the controls, the ultrastructure in the CN group had considerable variations. The axon of PE was swelled and deformed partly. The electron density was increased and presented as addicted to osmic acid. In the muscle cells, mitochondria was swelled, and sarcoplasmic reticulum was dilated. All PE exhibited ChAT immunoreactivity in human EOM. In the longitudinal section, nerve fibers extended from the muscle into the tendon, looped back and divided into several terminal arborizations (palisade endings) around the muscle fiber tip. The PE of medial rectus were richest at the location 3 - 4 mm from tendon insertion. In the cross section, the amount of PE in the CN group was higher than the control group (t = -5.613, P < 0.05). The expression level of ChAT in the CN group was higher than the control group (t = -3.730, P < 0.05). CONCLUSIONS: Palisade endings in myotendinous junction of human EOM are cholinergic nerves, which might innervate the contraction of EOM. Significant changes of palisade endings in the EOM of the CN subjects may affect eye movement.
