Slow PIII Research Articles

Hydrogen Peroxide Affects the Electroretinogram of Isolated Perfused Rat Retina

Purpose To evaluate the effects of H2O2 as an oxidant on the electroretinogram (ERG) in isolated rat retina. Methods Retinas were isolated from rat eyes and perfused with a nutrient solution. ERGs were recorded every 3 min. Once the signal was at a steady state, H2O2 was added to the perfusion solution. Results H2O2 caused instantaneous and transient changes in amplitudes and implicit times of the ERG, followed by changes in retinal survival curves. H2O2 0.2 mM produced a rapid increase in b-wave amplitude, followed by a return to the initial value and a survival curve above the control (without H2O2). A slight increase in a-wave was observed, followed by a decrease and a recovery above the control. The slow PIII decreased and then recovered to the initial value. H2O2 0.6 mM induced a small increase in b-wave amplitude, followed by a rapid decrease without recovery. The a-wave and slow PIII decreased rapidly without recovery. The implicit times of the a-wave and b-wave increased moderately with a low dose of H2O2, whereas they significantly increased with a high dose. Whatever the dose, the slow PIII implicit time increased significantly, followed by a return to the initial value. Barium increased the a-wave and b-wave, and then H2O2 reduced the two waves with a stronger effect on the a-wave. Aspartate and barium isolated the fast PIII, which decreased after H2O2 application. Conclusions H2O2 affects retinal function as shown by ERGs in isolated rat retina. The response differs with the dose of H2O2, suggesting that mechanisms underlying the action at low doses might be different from those at high doses. Our results also suggest an effect of H2O2 on ionic currents and/or neurotransmitter releases involved in the generation of the ERG and indicate a more pronounced effect on photoreceptors than on postsynaptic cells.

K+-dependent Müller cell-generated components of the electroretinogram.

The electroretinogram (ERG) has been employed for years to collect information about retinal function and pathology. The usefulness of this noninvasive test depends on our understanding of the cell sources that generate the ERG. Important contributors to the ERG are glial Müller cells (MCs), which are capable of generating substantial transretinal potentials in response to light-induced changes in extracellular K+ concentration ([K+]o). For instance, the MCs generate the slow PIII (sPIII) component of the ERG as a reaction to a photoreceptor-induced [K+]o decrease in the subretinal space. Similarly, an increase of [K+]o related to activity of postreceptor retinal neurons also produces transretinal glial currents, which can potentially influence the amplitude and shape of the b-wave, one of the most frequently analyzed ERG components. Although it is well documented that the majority of the b-wave originates from On-bipolar cells, some contribution from MCs was suggested many years ago and has never been experimentally rejected. In this work, detailed information about light-evoked [K+]o changes in the isolated mouse retina was collected and then analyzed with a relatively simple linear electrical model of MCs. The results demonstrate that the cornea-positive potential generated by MCs is too small to contribute noticeably to the b-wave. The analysis also explains why MCs produce the large cornea-negative sPIII subcomponent of the ERG, but no substantial cornea-positive potential.

Electroretinographic Assessment of Inner Retinal Signaling in the Isolated and Superfused Murine Retina

ABSTRACTPurpose: Longer-lasting electroretinographic recordings of the isolated murine retina were initially achieved by modification of a phosphate-buffered nutrient solution originally developed for the bovine retina. During experiments with a more sensitive mouse retina, apparent model-specific limitations were addressed and improvements were analyzed for their contribution to an optimized full electroretinogram (ERG).Material and methods: Retinas were isolated from dark-adapted mice, transferred to a recording chamber and superfused with different solutions. Scotopic and photopic ERGs were recorded with white flashes every 3 minutes. The phosphate buffer (Sickel-medium) originally used was replaced by a carbonate-based system (Ames-medium), the pH of which was adjusted to 7.7–7.8. Moreover, addition of 0.1 mM BaCl2 was investigated to reduce b-wave contamination by the slow PIII component typically present in the murine ERG.Results: B-wave amplitudes were increased by the pH-shift (pH 7.4 to pH 7.7) from 22.9 ± 1.9 µV to 37.5 ± 2.5 µV. Improved b-wave responses were also achieved by adding small amounts of Ba2+ (100 µM), which selectively suppressed slow PIII components, thereby unmasking more of the true b-wave amplitude (100.0% with vs. 22.2 ± 10.7% without Ba2+). Ames medium lacking amino acids and vitamins was unable to maintain retinal signaling, as evident in a reversible decrease of the b-wave to 31.8 ± 3.9% of its amplitude in complete Ames medium.Conclusions: Our findings provide optimized conditions for ex vivo ERGs from the murine retina and suggest that careful application of Ba2+ supports reliable isolation of b-wave responses in mice. Under our recording conditions, murine retinas show reproducible ERGs for up to six hours.

Early Functional and Morphologic Abnormalities in the Diabetic Nyxnob Mouse Retina.

PurposeThe electroretinogram c-wave is generated by the summation of the positive polarity hyperpolarization of the apical RPE membrane and a negative polarity slow PIII response of Müller glia cells. Therefore, the c-wave reduction noted in prior studies of mouse models of diabetes could reflect a reduction in the RPE component or an increase in slow PIII. The present study used a genetic approach to distinguish between these two alternatives.MethodsNyxnob mice lack the ERG b-wave, revealing the early phase of slow PIII. To visualize changes in slow PIII due to diabetes, Nyxnob mice were given streptozotocin (STZ) injections to induce diabetes or received vehicle as a control. After 1, 2, and 4 weeks of sustained hyperglycemia (>250 mg/dL), standard strobe flash ERG and dc-ERG testing were conducted. Histological analysis of the retina was performed.ResultsA reduced c-wave was noted at the 1 week time point, and persisted at later time points. In comparison, slow PIII amplitudes were unaffected after 1 week of hyperglycemia, but were significantly reduced in STZ mice at the 2-week time point. The decrease in amplitude occurred before any identifiable decrease to the a-wave. At the later time point, the a-wave became involved, although the slow PIII reductions were more pronounced. Morphological abnormalities in the RPE, including increased thickness and altered melanosome distribution, were identified in diabetic animals.ConclusionsBecause the c-wave and slow PIII were both reduced, these results demonstrated that diabetes-induced reductions to the c-wave cannot be attributed to an early increase in the Müller glia-derived potassium conductance. Furthermore, because the a-wave, slow PIII and c-wave reductions were not equivalent, and varied in their onset, the reductions cannot reflect the same mechanism, such as a change in membrane resistance. The presence of small changes to RPE architecture indicate that the c-wave reductions present in diabetic mice likely represents a primary change in the RPE induced by hyperglycemia.

Response properties of slow PIII in the Large vls mutant

Mouse mutants for proteins expressed in the dystrophin-glycoprotein complex at the photoreceptor terminal have electroretinogram (ERG) b-waves with a delayed onset and time course. The b-wave is defined by the sum of PII generated by depolarizing bipolar cells and slow PIII generated by Müller glial cells. In this study, we evaluated the hypothesis that the abnormalities observed in one of these mutants, Large (vls) , are caused by abnormal response properties of slow PIII. To isolate slow PIII, we crossed the Large (vls) mutant to a mouse line (Gpr179 (nob5) ) that lacks the ERG b-wave but maintains normal photoreceptor function and in which retinal degeneration does not occur. ERGs were recorded to strobe flash stimuli after overnight dark adaptation. In comparison with control responses, the a-wave and slow PIII had comparable waveforms but were reduced in amplitude in Large (vls) mice. The magnitude of this reduction was comparable for these components, and across stimulus luminance. There was no stimulus condition where the amplitude of slow PIII was larger than control. The data obtained are inconsistent with the idea that the b-wave abnormalities noted in Large (vls) mutant mice are caused by abnormal response properties of slow PIII.

Light-Evoked Responses of the Retinal Pigment Epithelium: Changes Accompanying Photoreceptor Loss in the Mouse

Mutations in genes expressed in the retinal pigment epithelium (RPE) underlie a number of human inherited retinal disorders that manifest with photoreceptor degeneration. Because light-evoked responses of the RPE are generated secondary to rod photoreceptor activity, RPE response reductions observed in human patients or animal models may simply reflect decreased photoreceptor input. The purpose of this study was to define how the electrophysiological characteristics of the RPE change when the complement of rod photoreceptors is decreased. To measure RPE function, we used an electroretinogram (dc-ERG)-based technique. We studied a slowly progressive mouse model of photoreceptor degeneration (Prph(Rd2/+)), which was crossed onto a Nyx(nob) background to eliminate the b-wave and most other postreceptoral ERG components. On this background, Prph(Rd2/+) mice display characteristic reductions in a-wave amplitude, which parallel those in slow PIII amplitude and the loss of rod photoreceptors. At 2 and 4 mo of age, the amplitude of each dc-ERG component (c-wave, fast oscillation, light peak, and off response) was larger in Prph(Rd2/+) mice than predicted by rod photoreceptor activity (Rm(P3)) or anatomical analysis. At 4 mo of age, the RPE in Prph(Rd2/+) mice showed several structural abnormalities including vacuoles and swollen, hypertrophic cells. These data demonstrate that insights into RPE function can be gained despite a loss of photoreceptors and structural changes in RPE cells and, moreover, that RPE function can be evaluated in a broader range of mouse models of human retinal disease.

Blockade of amiloride-sensitive sodium channels alters multiple components of the mammalian electroretinogram

Retinal neurons and Muller cells express amiloride-sensitive Na+ channels (ASSCs). Although all major subunits of these channels are expressed, their physiological role is relatively unknown in this system. In the present study, we used the electroretinogram (ERG) recorded from anesthetized rabbits and isolated rat and rabbit retina preparations to investigate the physiological significance of ASSCs in the retina. Based upon our previous study showing expression of alpha-ENaC and functional amiloride-sensitive currents in rabbit Muller cells, we expected changes in Muller cell components of the ERG. However, we observed changes in other components of the ERG as well. The presence of amiloride elicited changes in all major components of the ERG; the a-wave, b-wave, and d-wave (off response) were enhanced, while there was a reduction in the amplitude of the Muller cell response (slow PIII). These results suggest that ASSCs play an important role in retinal function including neuronal and Muller cell physiology.

Genetic Inactivation of an Inwardly Rectifying Potassium Channel (Kir4.1 Subunit) in Mice: Phenotypic Impact in Retina

The inwardly rectifying potassium channel Kir4.1 has been suggested to underlie the principal K(+) conductance of mammalian Müller cells and to participate in the generation of field potentials and regulation of extracellular K(+) in the retina. To further assess the role of Kir4.1 in the retina, we generated a mouse line with targeted disruption of the Kir4.1 gene (Kir4.1 -/-). Müller cells from Kir4.1 -/- mice were not labeled with an anti-Kir4.1 antibody, although they appeared morphologically normal when stained with an anti-glutamine synthetase antibody. In contrast, in Müller cells from wild-type littermate (Kir4.1 +/+) mice, Kir4.1 was present and localized to the proximal endfeet and perivascular processes. In situ whole-cell patch-clamp recordings showed a 10-fold increase in the input resistance and a large depolarization of Kir4.1 -/- Müller cells compared with Kir4.1 +/+ cells. The slow PIII response of the light-evoked electroretinogram (ERG), which is generated by K(+) fluxes through Müller cells, was totally absent in retinas from Kir4.1 -/- mice. The b-wave of the ERG, in contrast, was spared in the null mice. Overall, these results indicate that Kir4.1 is the principal K(+) channel subunit expressed in mouse Müller glial cells. The highly regulated localization and the functional properties of Kir4.1 in Müller cells suggest the involvement of this channel in the regulation of extracellular K(+) in the mouse retina.

Current source-density analysis of light-evoked field potentials in rabbit retina.

The technique of current source-density analysis was applied to several components of the light-evoked field potentials (electroretinogram) from the retina of the superfused eyecup of rabbit. The depth distributions of the major current sources and sinks were: b-wave--sink at outer plexiform layer, source at inner plexiform layer; M-wave--sink at inner plexiform layer, source at retinal surface; and slow PIII--source near outer plexiform layer, sink at retinal surface. These distributions, along with the sensitivities of these responses to certain pharmacological agents, support earlier studies that Müller cells generate the M-wave and slow PIII, but that depolarizing bipolar cells directly generate the b-wave.

Effects of bicarbonate ion on chick retinal pigment epithelium: membrane potentials and light-evoked responses.

The purpose of this study was to determine how changes in [HCO3-] alter the electrical properties of the retinal pigment epithelium (RPE). Experiments were conducted on the isolated chick retina-RPE-choroid preparation. The chamber holding the preparation allowed independent perfusion of the retinal and the choroidal surfaces. The light-evoked trans-tissue potential (TTP), the trans-epithelial potential (TEP), the trans-retinal potentials, and the intracellularly-recorded apical and basal membrane potentials were studied. Increasing the [HCO3-]0 in the choroidal bath from 25 to 40 mEq/1 led to an increase in the TTP and TEP. The same change in the retinal bath decreased the TTP because of a biphasic change of the RPE membrane potentials. There was also an increase in the amplitudes of the TEP, the c-wave and the slow PIII. The light-evoked subretinal K+ decrease was greater which is consistent with an increase in the photoreceptor light response. These observations indicated that the decrease of TTP resulted from a basal membrane hyperpolarization followed by an apical membrane depolarization induced by an increase in retinal [HCO3-]0. The relationship of these potential changes to the human bicarbonate responses is discussed.

Effects of Pituitary Adenylate-Cyclase-Activating Polypeptide on the Direct-Current Electroretinogram of the Rabbit Eye

Pituitary adenylate-cyclase-activating polypeptide (PACAP) is a recently discovered neuropeptide present in two different forms, PACAP-27 and PACAP-38. Both peptides stimulate the catalytic enzyme adenylate cyclase in pituitary cells. This enzyme is important also regarding the function of the retinal pigment epithelium (RPE). The purpose of the study was to investigate possible influences of PACAP on the rabbit retina and the RPE as reflected in the direct-current (d.c.) electroretinogram (ERG) and the standing potential of the eye (SP). After unilateral sector vitrectomy, a continuous intraocular perfusion with a reference solution alternated with a test solution was established. The corneal d.c. ERG and the SP were recorded from both eyes with the contralateral eye as a control. Both PACAP-27 (0.1 and 1 µM) and PACAP-38 (1 µM) increased the c-wave amplitude significantly (p = 0.028, p = 0.013 and p = 0.024, respectively, n = 4) while neither PACAP-27 (0.1 and 1 µM, p > 0.05, n = 4) nor PACAP-38 (1 µM, p > 0.05, n = 4) produced any significant effects on the a- and b-wave amplitudes of the d.c. ERG. The SP response to the two substances differed with a significant elevation of the SP level with PACAP-27 (1 µM, p = 0.017, n = 4), while PACAP-38 induced a small, nonsignificant SP elevation (1 µM, p > 0.05, n = 4). Retinal penetrations during PACAP-27 (10 µM) perfusion showed an increase in transepithelial potential (TEP) c-wave (p = 0.003) as well as in slow PIII (p = 0.011, n = 3) amplitude level. The results support the presence of PACAP receptors both on the RPE and in the neural retina.

Serum-induced changes in the physiology of mammalian retinal glial cells: role of lysophosphatidic acid.

1. With a breakdown of the vascular-CNS barrier, serum enters the nervous system. Although this is a frequent pathophysiological event, knowledge of the effects of serum on the function of the nervous system is limited. In this study, we examined the effects of serum on the activity of ion channels in Müller cells: the principal glia of the retina. 2. Freshly dissociated Müller cells from the bovine and human retina were studied with the perforated-patch configuration of the patch clamp technique. In other experiments, electroretinograms (ERGs) were recorded from isolated rat retinas. 3. Perforated-patch recordings revealed that serum induced a calcium-permeable, non-specific cation (NSC) current. Approximately 40 s after induction of this current, an outwardly rectifying K+ current was also detected. Sensitivity to charybdotoxin and margatoxin indicated that this K+ current was due to the activation of Kv1.3 channels. This increase in the Kv1.3 current was dependent on extracellular calcium. 4. The NSC and Kv1.3 currents were activated by serum in 100% and 95% of the sampled Müller cells, respectively. Also, in a minority (21%) of the cells, the inwardly rectifying K+ current was inhibited slightly. These changes in ion channel activity were associated with depolarization of the Müller cells. 5. We hypothesized that activation of NSC channels would reduce the siphoning of K+ via the Müller cells. Consistent with this idea, ERGs from isolated retinas showed serum-induced reductions in the slow PIII component, which is generated by Müller cells responding to light-evoked changes in the extracellular K+ concentration. 6. Lysophosphatidic acid (LPA), a component of serum, had effects on Müller cells that were qualitatively similar to those induced by serum. 7. Our observations demonstrate that exposure to serum alters the activity of multiple types of ion channels in Müller glial cells of the mammalian retina. When there is a breakdown of the blood-retina barrier, LPA may be one of the serum-derived molecules which regulates the physiology of Müller cells.

Neural retinal contribution to the slow negative potential of the canine electroretinogram.

To establish the normal waveform of the electrical responses from canine eyes, electroretinograms and the light peak were recorded in hybrid and beagle dogs under general anesthesia and artificial ventilation. The neural retinal and retinal pigment epithelial components were pharmacologically isolated by intravitreal glutamate injection and systemic sodium iodate administration, respectively. The a- and b-waves elicited by either flash or rectangular stimuli, the oscillatory potentials elicited by flash stimuli and the light peak elicited either by a single maintained illumination or by repetitive stimuli were almost identical with those of other vertebrates thus far studied. In contrast, in response to rectangular (several-second duration) stimuli, the c-wave was usually absent and was replaced by a slow cornea-negative potential that had a time course similar to that of the c-wave in other species. This slow negative potential was elicited at such low stimulus intensities that the a-wave was absent, was deepened by an intravenous administration of sodium iodate, was not affected by an intravitreal injection of sodium glutamate and was shallowed during the light peak. These results suggest that the slow negative potential that replaces the c-wave consists of large slow PIII, small retinal pigment epithelial c-wave and negligible contribution from the late photoreceptor potential and the dc component of the PII.

The origin of slow PIII in frog retina: current source density analysis in the eyecup and isolated retina.

The objective of this research was to determine the sources and sinks of current underlying the slow PIII component of the electroretinogram. Current source density analysis of the ERG evoked by diffuse light flashes was performed in eyecup and isolated retinas of frog. Blockade of synaptic transmission with aminophosphonobutyric + kynurenic acids simplified the CSD profiles through the retina. In addition to the photoreceptor source/sink pair, there was evidence for a major slow PIII source near the outer limiting membrane, a major sink near the inner limiting membrane, and a small source near the inner plexiform layer. Addition of Ba2+ abolished the slow PIII source/sinks, and it left only the photoreceptor source and sink. The results support the idea that slow PIII originates through K+ spatial buffering by Müller cells. Specifically, the light-evoked decrease in [K+]o in the subretinal space causes a primary K+ efflux from Müller cells (current source) and a primary K+ influx at the Müller cell endfeet (current sink). A decrease in [K+]zero in the proximal retina, caused by diffusion of K+ to the subretinal space, results in K+ efflux (the current source) at the inner plexiform layer.

Serotonin elevates the c-wave of the electroretinogram of the rabbit eye by increasing the transepithelial potential

The influence of serotonin (5-hydroxytryptamine, 5-HT) and serotonin analogues on the direct current electroretinogram (d.c. ERG) and the standing potential of the albino rabbit eye (SP) was studied. After unilateral vitrectomy, corneal recordings were obtained during simultaneous intravitreal perfusion with a control solution alternating with 5-HT at concentrations of 25, 120 and 200 μM. The c-wave increased at 25 and 120 μM when changing from control solution to test solution (P < 0.05) but did not decrease significantly when changing back to control solution (P > 0.05). The c-wave was reversibly elevated at 200 μM (PHS-5-HT, P < 0.01; 5-HT-PHS, P < 0.05). To analyse further the influence on the c-wave, in vivo intraretinal microelectrode recordings were obtained during intravitreal perfusion with 5-HT. The transepithelial potential (TEP) increased (P < 0.01), while the slow PIII was not significantly affected (P > 0.05). The serotonin receptor agonists 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane, 5-methoxytryptamine, α-methyl-5-hydroxytryptamine and 2-methyl-5-hydroxytryptamine, caused a significant reversible elevation of the c-wave, whereas 5-carboxyamidotryptamine did not. Tropisetron did not block the serotonin effect and LY53857 had an effect of its own on the c-wave. The results seem to indicate that the influence of serotonin on the c-wave is mainly due to an effect on the retinal pigment epithelium (RPE) and that more than one type of serotonin receptor may be involved.

Current-source density analysis of the electroretinogram of the frog: methodological issues and origin of components

The technique of current-source density (CSD) analysis for extracellular potentials is reviewed, along with some methodological features that are important for performing CSD analysis of the electroretinogram. In addition, three formulas for computing CSD's are examined on model circuits of resistors and current generators. Finally, CSD results from frog retina that bear on the origins of the b, d, and M waves, along with slow PIII, are presented. It is concluded that the b and d waves are generated primarily and directly by bipolar cells, whereas the M wave and the slow PIII are generated by Müller (glial) cells through the K+ spatial buffer mechanism.

A cyclic adenosine monophosphate agonist elevates the b- and c-waves of the rabbit direct-current electroretinogram.

The effects of the stable cyclic adenosine monophosphate analogue adenosine 3',5'-cyclic monophosphorothioate Sp-isomer (Sp-cAMPS) on the direct-current electroretinogram and the standing potential of the eye were studied. Corneal recordings were obtained from unilaterally vitrectomized albino rabbit eyes during alternating intravitreal perfusions with Sp-cAMPS and a control solution (Pharmacia eye irrigating solution). The contralateral eye was used as a control. To evaluate further the effects on the c-wave, in vivo intraretinal microelectrode measurements were made during simultaneous intravitreal perfusion of Sp-cAMPS and irrigating solution, respectively. Sp-cAMPS in concentrations of 1, 10 and 100 microM was tested by corneal direct-current electroretinography. There was no significant effect on the a-wave amplitude. The b-wave amplitude was reversibly elevated at an Sp-cAMPS concentration of 100 microM was tested by corneal direct-current electroretinography. There was no significant effect on the a-wave amplitude. The b-wave amplitude was reversibly elevated at an Sp-cAMPS concentration of 100 microM (p < 0.01, n = 7). The c-wave amplitude was reversibly elevated at an Sp-cAMPS concentration of 100 microM (p < 0.01, n = 7). The c-wave amplitude was reversibly elevated at a concentration of 10 microM (p < 0.001, n = 8), and this effect was more pronounced at 100 microM (p < 0.001, n = 7). The SP increased reversibly at a concentration of 100 microM (p < 0.001, n = 7). Microelectrode recordings were performed with Sp-cAMPS at a concentration of 100 microM. The recordings showed significant increases in both the transepithelial potential (p < 0.01, n = 3) and the slow PIII (p < 0.01, n = 3). The effects of Sp-cAMPS on the b-wave as well as the two components of the c-wave suggest influences on both the inner retina and the retinal pigment epithelium of the rabbit eye.

Adrenergic effects on the corneal and intraretinal direct-current electroretinogram and on the standing potential of albino rabbit eyes.

This study was undertaken to investigate further the responsiveness of the albino rabbit retinal pigment epithelium and the inner retina to adrenergic agents as reflected in changes of the direct-current electroretinogram and of the standing potential of the eye. After unilateral vitrectomy on albino rabbits, a continuous intraocular perfusion with a reference solution was established. The reference solution was then alternated with the test solution. The direct-current electroretinogram and the standing potential were recorded from both eyes with a scleral contact lens and a reference electrode connected to matched calomel half-cells. An in vivo experimental technique that allows intraocular perfusion of a test substance and simultaneous intraretinal microelectrode measurements was also used. The alpha-adrenergic agonist phenylephrine (0.04 microM, n = 8) produced a reversible increase in c-wave amplitude (48%, p < 0.001) and also a small increase in b-wave amplitude (12%, p < 0.002). There was no significant influence on the a-wave amplitude. The standing potential was elevated at 1694 +/- 362 microV (mean +/- SEM) (p < 0.002). The alpha 2-adrenergic agonist clonidine caused similar effects on the electroretinogram, although at a higher concentration (40 microM, n = 5), with an elevation of the c-wave (25%, p < 0.001) and a small b-wave increase (12% p < 0.002). No significant influence on the a-wave or on the standing potential was found. Intraretinal direct-current electroretinogram-recordings during intraocular perfusion with phenylephrine showed an increase in transepithelial potential (p < 0.004; n = 6), accompanied by a reduction of the slow PIII (p < 0.0035; n = 6). The c-wave increase resulting from alpha-adrenergic stimulation seems to be generated partly across the retinal pigment epithelium, with an increase in transepithelial potential, combined with a reduction of the slow PIII. The elevation of the b-wave amplitude, together with the influence on the slow PIII, suggests alpha-adrenergic effects also on the inner retina. The experimental technique used in this study with intraocular perfusion after vitrectomy and simultaneous intraretinal direct-current recordings seems to be a practicable method for studies of the influence of pharmacologic agents on the retina and the retinal pigment epithelium.

Effects of PhXA41, a prostaglandin analogue, and PGF2 alpha on the corneal and intraretinal d.c. electroretinogram (ERG) of the albino rabbit eye.

PhXA41, a prostaglandin F2 alpha analogue, is a new drug alternative for medical treatment of glaucoma. From earlier studies, it is known that certain prostaglandins (pgs) exert an influence on the d.c. electroretinogram (ERG). In the present study, possible effects of intraocular perfusion with PhXA41 (free acid) or PGF2 alpha on the rabbit retina and retinal pigment epithelium (RPE) were evaluated. Unilateral vitrectomy was performed and a continuous intraocular perfusion was established, in which a reference solution was alternated with the test solution. In part one of the experiments, the corneal d.c. ERG and the standing potential of the eye (SP) were recorded from both eyes. In part two, vitreal and intraretinal d.c. ERG were recorded in the vitrectomized eye. PhXA41 and PGF2 alpha produced similar effects, i.e. a significant and reversible c-wave increase at concentrations of 1-10 micrograms/ml. No influence was found in the concentration interval 10-100 ng/ml. There were no changes in a- and b-wave amplitudes at any of the concentrations tested. At a concentration of 10 micrograms/ml, the SP increased significantly. The intraretinal d.c. ERG showed a large reduction of slow PIII accompanied by a small change in transepithelial potential (TEP). Theoretically, a vitreous concentration of less than 1 ng/ml may be reached after topical application. This concentration level does not influence the d.c. ERG in this animal model. At higher concentrations, a significant and reversible c-wave increase seemed to be generated mainly in the inner retina by a reduction of slow PIII generated in the Müller cells. The effect of PhXA41 appears to be mediated by an FP receptor.

The c-wave of the electroretinogram possesses a third component from the proximal retina

ERG and light-induced extracellular potassium ([K +] o) changes have been measured in isolated retinas of both Rana esculenta and Rana temporaria. The conditions of the preparations have been varied. Isolated frog retinas kept receptor side-upward in a moist chamber without perfusion showed the well-known slow PIII in the ERG. Retinas superfused from the receptor side, with O 2 enrichment at their vitreal surface, however, exhibit a slow cornea-positive potential in the ERG. The slow ERG-potentials relate to different light-induced potassium changes in the proximal retina. There was a long lasting and larger proximal potassium increase in adequately maintained retinas but a smaller and shorter one in preparations lacking superfusion and oxygen. There was no significant difference between the size of potassium decrease around receptors of retinas supervised from their vitreal side and those supervised from receptor side. A reduction of slow PIII should therefore not be responsible for the slow cornea-positive potential. The long lasting and larger (by 59%) potassium increase in the proximal retina may counteract the potential in the Mu¨ller cells caused by the potassium decrease around receptors and thereby cancel slow PIII and generate a third component of the electroretinogram c-wave.