Effects of light and darkness on pH outside rod photoreceptors in the cat retina.

We recorded pH in the extracellular space surrounding rod photoreceptors in the dark-adapted eye of the cat and during illumination with double-barreled H(+)-selective microelectrodes. A pH of 7.17 was recorded in the vitreous at the retinal surface of the dark-adapted eye and this became more alkaline during light adaptation. In dark adaptation, a pH close to 7.00 was recorded in a region of maximal acidity in the extracellular space surrounding rods in the outer nuclear layer (ONL). pH steeply alkalinized as the microelectrode was moved more distally towards the retinal pigment epithelium (RPE), and almost reached the pH of the arterial blood at the apical surface of the RPE. Illumination produced an intraretinal alkalinization that was largest (up to 0.2 pH units) in the ONL, maximal in amplitude at rod-saturating intensities, and that was sustained during steady background illumination. The light-evoked alkalinization was relatively slow in onset, having a time constant (1/e) of 64 sec, and took 8-12.5 min to return to the dark-adapted level after the offset of maintained illumination. These results show that acid production by cat rods is highest in the dark, reflecting a high rate of energy metabolism, and suggest that glycolysis is required to support the dark current. Illumination, by suppressing both glycolysis and respiration, alkalinizes the extracellular space surrounding rods. The substantial change in pH outside rods from dark to light could alter pH dependent properties of the interphotoreceptor matrix.

Light-evoked changes in extracellular pH in frog retina.

Light-induced changes in extracellular H+ concentration (delta pH0) were studied with intraretinal H(+)-sensitive double-barreled microelectrodes in frog eyecup and isolated retina preparations. The most prominent delta pH0 were found in the inner plexiform layer, as pH increases (alkalinizations) at light onset and offset. With a small-spot stimulus (0.3 mm dia.), 30 sec in duration, the delta pH0 were relatively small (0.03 pH units), and long lasting (peak at 25-30 sec). They were enhanced by flicker (0.3 Hz). Depth profiles paralleled those of the field potentials (PNR/M-wave), the ON delta pH0 peaking 40 microns more proximal than the OFF response. The delta pH0 exhibited surround antagonism, which was blocked by tetrodotoxin (TTX), indicating an independence from action potentials. The mechanism for these pH increases in proximal retina is not yet understood. In the subretinal space diffuse retinal illumination produced a small pH increase, consistent with a presumed decrease in photoreceptor lactate production. Inhibition of carbonic anhydrase (CA) with acetazolamide or methazolamide increased both the proximal and distal retinal delta pH0, suggesting that CA is involved in buffering retinal pH.

Retinal detachment in the cat: the pigment epithelial-photoreceptor interface.

Twenty-six cat retinae were surgically detached by injecting fluid into the subretinal space (SRS). The retinae were then studied by light and electron microscopy at detachment intervals ranging from 1/2 hr to 14 months. Degenerative and proliferative changes occur at the retinal pigment epithelial (RPE)-photoreceptor interface very soon after detachment, and the severity of these changes depends upon both the duration and height of the detachment. The specialized apical RPE processes that ensheath the outer segments are replaced by a uniform fringe of short, undifferentiated processes. The apical RPE surface becomes mounded, and this mounding becomes more pronounced at longer detachment durations. Labeling experiments with 3H-thymidine showed that some cat RPE cells enter a phase of stimulated DNA synthesis 12-24 hrs after detachment; RPE mitotic figures are first apparent 48 hrs after detachment. In the cat, discrete regions of proliferated RPE cells usually appear in one of several configurations. A number of different cell types, including polymorphonuclear neutrophils, monocytes at various maturational stages, photoreceptor cells, Müller cells, and RPE cells, appear in the expanded SRS of detached retinae. Rod and cone outer segments degenerate rapidly and become membrane bound sacs by 3 days postdetachment; the assembly of new outer segment membrane apparently does not stop completely even at moderately long detachment intervals (ie, 2 months). Degenerative changes in the inner segments do not take place with the same rapidity as those in the outer segments. The changes that occur at the RPE-photoreceptor interface are rapid, progressive, and sometimes irreversible events that have significant implications for photoreceptor recovery following retinal reattachment surgery.

Retinal detachment in the cat: the outer nuclear and outer plexiform layers.

The retinae of cats were surgically detached for 1/2 hr to 14 months, and the outer nuclear (ONL) and outer plexiform layers (OPL) were studied by light and electron microscopy. The longer the duration or the greater the height of detachment the more likely was the occurrence of cell death. Histologic signs of degeneration were present 1 hr after detachment. The number of photoreceptor nuclei in the ONL decreased significantly by 1 month. Loss of cells in the ONL occurred by necrosis and by the migration of photoreceptor cell bodies into the subretinal space. The OPL degenerated by the necrosis of cell processes and synaptic terminals and by the retraction of the synaptic terminals. By 2 weeks most synaptic terminals were necrotic or in the process of retracting. Photoreceptor synaptic contact with second order neurons was diminished by 30 days and was essentially absent by 50 days. Müller cells proliferated and hypertrophied; their nuclei and cell processes filled the intraretinal spaces left by the degenerating photoreceptors. In addition, Müller cells protruded into the subretinal space and formed multiple layers of cell bodies and processes between the retina and retinal pigment epithelium. By 14 months these subretinal Müller cell processes covered the entire detached retina, and appeared morphologically like an astroglial scar. Similar changes in human retinal detachments may significantly influence the degree of visual recovery after retinal reattachment, especially in retinae detached for more than a few days.

Epiretinal membrane formation after vitrectomy.

Our experimental model of epiretinal membrane formation in the rabbit eye after lensectomy and vitrectomy provides a way of studying pharmacologic and surgical approaches to inhibiting epiretinal cellular proliferation and contraction in the eye that has undergone vitrectomy. We injected 400,000 tissue-cultured retinal pigment epithelial cells onto the retinal surface of rabbit eyes that had undergone lensectomy, vitrectomy, and fluid-gas exchange. By one week, a funnel-shaped detachment of the medullary rays had occurred in 100% of the injected eyes. Histologically, the cells formed an epiretinal membrane by six hours after injection and caused major wrinkling of the inner retina after 24 to 48 hours. The percentage of tritiated-thymidine-labeled epiretinal cells increased dramatically 24 hours after injection and then declined. Cellular membranes bridging the optic nerve, followed by growth and contraction of the epiretinal cells on the detached internal limiting membrane, were responsible for the closed funnel appearance of the medullary rays.

The onset of pigment epithelial proliferation after retinal detachment.

The adult mammalian retinal pigment epithelium (RPE) is mitotically inactive, yet retains the capacity to proliferate under certain conditions. To determine the onset of RPE proliferation after retinal detachment, we examined experimentally detached cat retinas of 12, 24, 48, and 72 hr duration. An additional animal served as a nondetached, sham-operated control. 3H-thymidine was injected into the vitreous chamber and the eyes were processed for light microscopic autoradiography. Autoradiograms from both the control and the 12 hr detachment showed no evidence of labeled RPE nuclei; however, labeled nuclei were present at both 24 and 48 hr after detachment. Labeled nuclei per millimeter of RPE at 24 hr were 55% of the 48 hr rate. Mitotic figures were noted only at 48 and 72 hr after detachment. No labeled RPE nuclei appeared in autoradiograms that bordered the detachment zone. Electron micrographs showed that proliferating RPE cells assume several configurations, some of which have been reported in other species. The proliferative response of the RPE occurs much sooner than had previously been thought. It appears to be a local effect that does not involve retinal regions beyond the detachment boundaries, and it may have potentially adverse effects when the retina and RPE are reapposed after retinal reattachment surgery.

Anatomical recovery following retinal detachment: clinicopathological correlations.

The histological findings in experimental retinal detachment and reattachment of the cat retina are described. Loss of photoreceptor outer segments, mounding of the apical surface of the retinal pigment epithelium, and loss of the apical villi of the retinal pigment epithelium are prominent features of the detached cat retina. Reattachment within the first week of retinal detachment is followed by good photoreceptor regeneration and orientation. Longer term detachment of 2 to 6 weeks results in poorer photoreceptor outer segment regeneration and orientation 4 weeks following reattachment surgery. These experimental results are consistent with clinical data concerning visual recovery following retinal reattachment surgery.