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.

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 Nyxnob 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 (RmP3) 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 Müller 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 α-ENaC and functional amiloride-sensitive currents in rabbit Müller cells, we expected changes in Müller 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 Müller cell response (slow PIII). These results suggest that ASSCs play an important role in retinal function including neuronal and Müller cell physiology.

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.

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+]0 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+]0 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.

M-wave of the toad electroretinogram

1. In the retina, two distinct, light-evoked releases of K+ have been described. One takes place in the outer plexiform layer (OPL) and is termed the "distal K+ increase." The other takes place in the inner plexiform layer (IPL) and is termed the "proximal K+ increase." Although the distal K+ increase generates the electroretinogram (ERG) b-wave, the contribution of the much larger proximal K+ increase to the ERG is less well understood. In this paper we detail our investigation of the proximal K+ increase and its contribution to the ERG. We describe an ERG component, the M-wave, which had not heretofore been observed in the diffuse-flash, vitreal ERG. 2. We studied the proximal K+ increase and the ERG M-wave in the isolated retina preparation of the toad, Bufo marinus. We used K(+)-sensitive microelectrodes, as well as conventional intra- and extracellular microelectrodes, to record K+ changes, the local (or intraretinal) ERG, the vitreal ERG, and Muller cell responses. 3. As in earlier studies of the amphibian and cat M-wave, we readily observed an M-wave in the intraretinal, or local, ERG (LERG). The M-wave we studied had characteristics similar to those of M-waves that were previously described. Specifically, we found that the M-wave was generated by a Muller cell response to the proximal K+ increase and that both the proximal K+ increase and the LERG M-wave were spatially tuned. 4. We used the aspartate receptor agonist, N-methyl-DL-aspartate (NMA), to reveal that an M-wave is present in the vitreal ERG. Researchers who previously investigated the M-wave were unable to identify an M-wave in the vitreal ERG. We found that the toad ERG M-wave was a small, positive potential that was partially obscured by the much larger b-wave and slow PIII components. 5. We observed that picrotoxin (PTX) had an excitatory effect on inner retina, as evidenced by an enhanced proximal K+ increase and an enhanced M-wave. This result indicates that it is likely that GABAergic inhibition in inner retina plays an important role in retinal processing in the toad. 6. At threshold, we found that the ERG consisted mainly of an M-wave, indicating that the amphibian threshold ERG is driven by proximal retina. This result is analogous to previous observations of the threshold ERG in cat. However, in cat, the M-wave and threshold response have been described as distinct ERG components.(ABSTRACT TRUNCATED AT 400 WORDS)

Origin of negative potentials in the light-adapted ERG of cat retina

1. The light-adapted diffuse-flash electroretinogram (ERG) in the cat exhibits two prominent negative components: a sustained negative potential during illumination and a negative-going OFF response. We investigated their intraretinal origins and found that the sustained component originates from the rod photoreceptors, whereas the OFF response represents a combination of the return to base line of the rod-receptor potential, the offset of PII (rod and cone), and a cone-dependent OFF response originating proximal to the photoreceptors at very high background levels. 2. The ERG, evoked in response to diffuse illumination of the light- and dark-adapted cat retina, was recorded between a chlorided silver wire in the vitreous and a plate behind the eye. Extracellular field potentials were recorded simultaneously with a microelectrode placed intraretinally at different retinal depths. 3. The sustained negative potential and the negative OFF response were not the M-wave ON and OFF responses of proximal retina, despite an overall resemblance in form and time course: 1) the M-wave was spatially tuned, whereas the ERG components were not; 2) tetrodotoxin (TTX) (3.8-microM vitreal concentration) did not alter the M-wave, but it reduced the ERG OFF response; 3) picrotoxin (0.14 mM, after TTX) enhanced the M-wave but did not affect the negative ERG; and 4) 2-amino-4-phosphonobutyric acid (APB; 0.95 mM) removed the M-wave ON response, and aspartate (43 mM) removed the M-wave OFF response, in addition, while the sustained negative potential persisted. 4. The sustained negative potential was not slow PIII, the neural retinal component of the c-wave, a Muller cell response to the photoreceptor-dependent light-evoked decrease in subretinal extracellular K+ concentration [( K+]o). Although Ba2+ (repeated injections of 4-5 mM), a K+ conductance blocker, eliminated slow PIII, it did not remove the sustained negative potential. We concluded that the sustained negative potential was a photoreceptor potential, and the spectral sensitivity of the response indicated that it arose from rods. 5. The contribution of the rod-receptor potential to the ERG depended on background illumination. It was a sustained potential for a range of backgrounds near and 1 or 2 log units above the illumination that saturates rod-driven responses in cat (8.2 log quanta.deg-2.s-1). At lower background intensities, it appeared only as a dip between the b- and c-waves, the b-wave trough, which also was present in fully dark-adapted responses.(ABSTRACT TRUNCATED AT 400 WORDS)