The morphologic characteristics of corneal nerves in advanced keratoconus as evaluated by acetylcholinesterase technique.

PURPOSE: To study the morphologic characteristics of corneal nerves in patients with advanced keratoconus using the acetylcholinesterase technique in corneal whole mounts. DESIGN: Prospective, observational case series. METHODS: Fourteen corneal buttons from 14 keratoconic patients (9 males and 5 females; mean age, 34.3 years) who had undergone keratoplasty for advanced keratoconus and 6 corneal buttons from 6 normal corneas were included. Whole mounts were stained for acetylcholinesterase and were scanned with a novel digital pathology scanning microscope. RESULTS: Seventy-one percent of keratoconic corneas demonstrated central stromal nerve changes, which included thickening, tortuosity, nerve spouting, and overgrowth. The nerve changes ranged from early to extensive and could be separated into 3 different grades. The central stromal nerves were abnormally thicker (18.9 ± 14.7 µm) than in controls (8.11 ± 3.31 µm; P < .001). The thickness of peripheral stromal nerves (12.6 ± 3.1 µm) was similar to that of controls (14.86 ± 5.60 µm; P = .072). Subbasal nerves showed changes in the form of loss of radial orientation and increased tortuosity, especially at the cone apex. At the cone base, a concentric arrangement of subbasal nerves was found in 43% of cases. Localized thickenings of subbasal nerves also were observed at their origin from the bulbous terminations of sub-Bowman nerves. The terminal bulbs, too, were enlarged. The mean diameter of the subbasal nerves in keratoconus (4.11 ± 0.60 µm) did not differ from that of the controls (4.0 ± 0.61 µm; P = .422). CONCLUSIONS: This study provides additional histologic evidence of the involvement of corneal nerves in keratoconus and suggests further that they may play a role in the pathophysiologic factors and progression of the disease.

Immunohistochemical determination of the sympathetic pathway in the orbit via the cranial nerves in humans.

OBJECT: The present study was undertaken to elucidate the extent and precise distribution of the postganglionic sympathetic fibers in the cranial nerves projecting to the orbit and to reconstruct sympathetic routes in the orbit in humans. For this purpose, the authors made an immunohistochemical determination of the sympathetic fibers by using an antibody against norepinephrine-synthetic enzyme, tyrosine hydroxylase (TH). METHODS: Specimens containing the orbit and the cavernous sinus were obtained from formalin-fixed human cadavers. First, it was confirmed that the superior cervical ganglion contained strongly immunostained TH-positive neuronal cell bodies and fibers. After careful dissection of the cranial nerves projecting to the orbit, different segments of each cranial nerve were processed for immunohistochemical analysis for TH. All of the intraorbital cranial nerves contained TH-positive sympathetic fibers, although the amounts were very different in each cranial nerve. At the proximal site of the common tendinous ring, TH-positive fibers were found mainly in the abducent and trochlear nerves. At the distal site of this ring, TH-positive fibers were lost or markedly reduced in number in the abducent and trochlear nerves and were distributed mostly in the ophthalmic and oculomotor nerves. Among the cranial nerves projecting to the orbit, the ophthalmic nerve and its bifurcated nerves--frontal, lacrimal, and nasociliary--contained numerous TH-positive fibers. CONCLUSIONS: The authors conclude that the postganglionic sympathetic fibers are distributed to all cranial nerves projecting to the orbit and that the ophthalmic nerve provides a major sympathetic route in the orbital cavity in humans.

Immunohistochemical analysis of neuronal nitric oxide synthase in the trigeminal ganglia of the crotaline snake Trimeresurus flavoviridis.

The expression of neuronal nitric oxide synthase (nNOS) in the trigeminal ganglia (TG) of the infrared-sensitive crotaline snake Trimeresurus flavoviridis was studied immunohistochemically. The percentage of nNOS-positive (+) neurons in the TG was significantly higher (about 3.5-fold, P<0.001) in the mandibular division than in the infrared-sensory processing area (the maxillary division and ophthalmic ganglion). nNOS was found in varying sizes of TG neurons. However, nNOS (+) neurons were more abundant in small and large neurons than in medium-sized neurons, which include most of the infrared-sensitive neurons of the TG. These findings suggest that nNOS may be involved in normal physiological functions, such as the transmission of tactile, vibrotactile, and nociceptive sensations in the TG, rather than in infrared sensory processing in this species.