Contractile markers distinguish structures of the mouse aqueous drainage tract.

PURPOSE: Structures of the aqueous humor drainage tract are contractile, although the tract is not entirely composed of muscle. We characterized the mouse aqueous drainage tract by immunolabeling contractile markers and determined whether profiling these markers within the tract distinguished its key structures of the trabecular meshwork (TM) and ciliary muscle (CM). METHODS: Enucleated eyes from pigmented C57BL/6 (n=8 mice) and albino BALB/c (n=6 mice) mice were processed for cryo- and formalin-fixed paraffin-embedded sectioning. Immunofluorescence labeling was performed for the following: (a) filamentous actin (using fluorescence-conjugated phalloidin), representing a global contractile marker; (b) α-smooth muscle actin (α-SMA), caldesmon, and calponin, representing classic smooth muscle epitopes; and (c) nonmuscle myosin heavy chain, representing a nonmuscle contractile protein. Tissue labeling was identified by confocal microscopy and analyzed quantitatively. Hematoxylin and eosin staining provided structural orientation. RESULTS: A small portion of the TM faced the anterior chamber; the rest extended posteriorly alongside Schlemm's canal (SC) within the inner sclera. Within the drainage tract, filamentous actin labeling was positive in TM and CM. α-SMA and caldesmon labeling was seen primarily along the CM, which extended from the anterior chamber angle to its posterior termination beyond the SC near the retina. Low intensity, patchy α-SMA and caldesmon labeling was seen in the TM. Myosin heavy chain immunoreactivity was primarily found in the TM and calponin was primarily observed in the CM. C57BL/6 and BALB/c comparison showed that pigment obscured fluorescence in the ciliary body. CONCLUSIONS: Our strategy of profiling contractile markers distinguished mouse aqueous drainage tract structures that were otherwise indistinguishable by hematoxylin and eosin staining. The mouse TM was seen as an intervening structure between SC, a part of the conventional drainage tract, and CM, a part of the unconventional drainage tract. Our findings provide important insights into the structural and functional organization of the mouse aqueous drainage tract and a basis for exploring the role of contractility in modulating aqueous outflow.

Reversal of disc cupping after intraocular pressure reduction in topographic image series.

PURPOSE: To identify and characterize 'reversal' of optic nerve cupping following intraocular pressure (IOP) lowering in scanning laser tomography (SLT) longitudinal image series. METHODS: Modification was made to a previously described analytical approach to longitudinally study putatively increased rim area following IOP lowering. Sustained IOP reduction of 25% was by topical medication. Forty SLT image series with equivalent follow up were assessed: 10 with ocular hypertension (OHT), 10 with primary open angle glaucoma (POAG), and as controls, 20 normal. Reproducible rim area reversal was identified by sector and its time-course over 1 year examined. RESULTS: By a 2-of-3 reproducibility criterion, reversal following IOP lowering was confirmed in about a third of treated eyes (POAG and OHT) but not in any controls. Rim sectors showing reversal were mostly nasal, with a few occurring superotemporally. Reversal in a fifth of treated eyes persisted for at least 1 year; all these were in the nasal half of the disc. The number of sectors with persisting reversal affected less than 6% of all treated eyes' rim sectors. CONCLUSION: Rim area is not uncommonly increased after IOP lowering and this 'reversal' may persist for at least a year. Within topically treated eyes having IOP lowering of at least 25%, the proportion of rim sectors with persistent reversal appears small. Nevertheless, the effects of IOP reduction on topography, especially in the short term, should be considered when longitudinally assessing progressive rim loss in SLT images.