Innervation of extraocular pulley smooth muscle in monkeys and humans.

PURPOSE

Soft pulleys stabilize paths and determine pulling directions of the extraocular muscles (EOMs). This study was conducted to characterize innervation of smooth muscles (SMs) supporting these pulleys.

METHODS

Cadaveric human and monkey orbits were step and serially sectioned for histochemical and immunohistochemical staining. Before perfusion, the superior cervical ganglia of one monkey had been injected with the anterograde tracer Phaseolus vulgaris leukoagglutinin (PHA-L). Immunoperoxidase staining to human SM alpha-actin confirmed pulley SM. Monoclonal and polyclonal antibodies were used to demonstrate PHA-L, tyrosine hydroxylase, dopamine beta-hydroxylase, phenylethanolamine-N-methyltransferase, neuronal nitric oxide synthase (NOS), and synaptophysin. The NADPH-diaphorase reaction was also used as a marker for NOS and the acetylcholinesterase (AChE) reaction for acetylcholine.

RESULTS

Pulleys, consisting of collagen and elastin sleeves supported by connective tissue containing SM, were observed around rectus muscles of humans and monkeys. The human and monkey SM was richly innervated. Axons terminating in motor end plates within SM bundles were immunoreactive to PHA-L, tyrosine hydroxylase, and dopamine beta-hydroxylase, but not phenylethanolamine-N-methyltransferase, indicating innervation of pulley SM from the superior cervical ganglion by projections using norepinephrine. Smaller axons and motor end plates were also demonstrated in SM, using NADPH-diaphorase and NOS immunoreactivity, indicating nitroxidergic innervation, and using AchE, indicating cholinergic parasympathetic innervation. The pterygopalatine and, to a lesser extent, the ciliary ganglia, but not the Edinger-Westphal nucleus, contained cells immunoreactive to NOS, suggesting that nitroxidergic innervation to pulley SM is mainly from the pterygopalatine ganglion.

CONCLUSIONS

The SM suspensions of human and monkey EOM pulleys are similar and receive rich innervation involving multiple neurotransmitters. These complex projections suggest excitatory and inhibitory control of EOM pulley SM, and support their dynamic role in ocular motility.