Adaptation in the isolated rat retina.
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PURPOSE: Laser Doppler measurements performed immediately after the transition from dark adaptation (DA) to light led to the hypothesis that retinal blood flow, Fret, is increased during DA, but the use of visible lasers had prevented measurements during DA. Our aim was to test this hypothesis by measuring Fret during and after DA. MATERIAL AND METHODS: Fret in retinal vessels at the optic disk surface was recorded quasi-continuously in one eye of 6 normal subjects (age 27 to 60 years) using a laser Doppler flowmeter in the near-infrared (810 nm). Measurements were performed during light (baseline), various periods of DA and again during light. DA lasted between 2 and 32 min. RESULTS: Average Fret for the 6 subjects did not change significantly (-2.7 +/- 8% sd, p > 0.05) during the various periods of DA, as determined from linear regressions of the flux versus time. Following the transition from DA to light, there was, in most cases, a rapid transient increase of the flux, which reached an average value of 37 +/- 10% above the pre-transition value and peaked at 30-60 sec after the transition. CONCLUSIONS: These results do not support the hypothesis that Fret in normal volunteers is increased during DA. Rather, they strongly suggest that the transient increase in flux observed after DA is induced by the transition from dark to light (FNSRS #3200-043157 et CNR, It. #95.01715.CT04).
Phosducin is a photoreceptor-specific protein known to interact with the beta gamma subunits of G proteins. In pursuit of the function of phosducin, we tested the hypothesis that it regulates the light-driven translocation of G protein transducin from the outer segments of rod photoreceptors to other compartments of the rod cell. Transducin translocation has been previously shown to contribute to rod adaptation to bright illumination, yet the molecular mechanisms underlying the translocation phenomenon remain unknown. In this study we provide two major lines of evidence in support of the role of phosducin in transducin translocation. First, we have demonstrated that transducin beta gamma subunits interact with phosducin along their entire intracellular translocation route, as evident from their co-precipitation in serial tangential sections from light-adapted but not dark-adapted retinas. Second, we generated a phosducin knockout mouse and found that the degree of light-driven transducin translocation in the rods of these mice was significantly reduced as compared with that observed in the rods of wild type animals. In knockout animals the translocation of transducin beta gamma subunits was affected to a larger degree than the translocation of the alpha subunit. We also found that the amount of phosducin in rods is sufficient to interact with practically all of the transducin present in these cells and that the subcellular distribution of phosducin is consistent with that of a soluble protein evenly distributed throughout the entire rod cytoplasm. Together, these data indicate that phosducin binding to transducin beta gamma subunits facilitates transducin translocation. We suggest that the mechanism of phosducin action is based on the reduction of transducin affinity to the membranes of rod outer segments, achieved by keeping the transducin beta gamma subunits apart from the alpha subunit. This increased solubility of transducin would make it more susceptible to translocation from the outer segments.
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The S-potentials recorded intracellularly from the all-rod retina of the skate probably arise from the large horizontal cells situated directly below the layer of receptors. These cells hyperpolarize in response to light, irrespective of stimulus wavelength, and the responses in photopic as well as scotopic conditions were found to be subserved by a single photopigment with lambda(max) = 500 nm. The process of adaptation was studied by recording simultaneously the threshold responses and membrane potentials of S-units during both light and dark adaptation. The findings indicate that the sensitivity of S-units, whether measured upon steady background fields or in the course of dark adaptation, exhibits changes similar to those demonstrated previously for the ERG b-wave and ganglion cell discharge. However, the membrane potential level of the S-unit and its sensitivity to photic stimulation varied independently for all the adapting conditions tested. It appears, therefore, that visual adaptation in the skate retina occurs before the S-unit is reached, i.e., at the receptors themselves.
Rods and cones in Necturus respond with graded hyperpolarization to test flashes spanning about 3.5 log units of intensity. Steady background levels hyperpolarize the rods, and the rod responses become progressively smaller as background level is increased. In cones, higher background levels reduce the effectiveness of test flashes, so higher ranges of test intensities are required to elicit the full range of graded responses. When backgrounds are terminated, cones return rapidly, but rods return slowly to the dark potential level. The effects of backgrounds on both rods and cones can be observed at intensities that cause negligible bleaching as determined by retinal densitometry. During dark adaptation, changes are observed in the rods and cones that are similar to those produced by backgrounds. Receptor sensitivities, derived from these results, show that rods saturate, cones obey Weber's law, and sensitization during dark adaptation follows a two-phase time-course.
The proximal negative response (PNR), a complex extracellular potential derived mainly from amacrine cell activity, was studied in the all-rod retina of the skate. Tetrodotoxin (10(-6) mg/ml) did not affect either the waveform or the latency of the response, indicating that the PNR reflects the graded, nonregenerative components of the amacrine cell potential. As regards its adaptive properties, the PNR exhibited both the extreme sensitivity to weak background light and the slow time course of light and dark adaptation that are characteristic of other responses from the proximal retina. Thus, the PNR, like the b-wave and ganglion cell discharge, appears to reflect adaptive processes located within the neural network of the inner retina.
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At threshold the eye is known to sum quanta as though the total catch were important and not the distribution on the retina. In humans, spatial summation is larger under dark adapted conditions. The extent of this spatial summation is studied in goldfish retinal ganglion cells which are known to receive both rod and cone inputs. The area of summation in single ganglion cells is found to be much larger for rod input compared to cone input. It seems that in single ganglion cells, the spatial summating properties increase markedly in the dark due largely to the neural organization of rod and cone input.
The patterned visually evoked potential (pVEP) and patterned electroretinogram (pERG) after light and dark adaptation were investigated in 10 visually normal human observers. The influence of retinal preadaptation on the pVEP appeared to be dependent on the stimulus field size. With a 6 degree field size retinal adaptation effects were not apparent, but with a 12 degree field size the amplitudes of the dark-adapted steady-state pVEP (ssVEP) and transient pVEP (tVEP) were significantly larger than the light-adapted amplitudes. Similarly pERG's elicited by a 12 degree field size produced larger amplitude dark-adapted results. The implications of these findings are discussed. Monitoring the temporal changes in the pVEP subsequent to different retinal preadaptation levels proved to be inconclusive for the conditions studied.
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1. Low-frequency light-sensitive membrane current noise in isolated rod photoreceptors of the larval tiger salamander was recorded using suction electrodes, in the dark, and during light adaptation by backgrounds or by bleaching visual pigment. 2. In background light, noise variance increases and then decreases. For rods desensitized to similar levels by bleaching visual pigment, the noise variance either does not change (weak adaptation) or decreases (with stronger adaptation). 3. The power spectral density of the current noise in dark-adapted rods shows a component with half-power cut-off frequency at about 0.1 Hz, attributed to spontaneous single events and continuous noise from dark phosphodiesterase activity. A second component, with half-power cut-off frequency at about 1 Hz, may be due to slow components in the light-sensitive channel gating. 4. The power spectral density of the noise in background light is dominated by noise generated by the background. Background light adapts at least the first component of the noise seen in dark-adapted cells. For cells desensitized by bleaching, light adaptation of both components of the dark-adapted noise is observed. 5. The results confirm that the low-frequency noise in dark-adapted cells arises from the transduction mechanism of the rod, in that both components can be light adapted, and show that, for rods permanently desensitized by bleaching, the desensitization is not due to the presence of active visual pigment molecules similar to those produced by background light.
1. We have examined the responses of neurones in the suprachiasmatic nuclei (SCN) of the rat to retinal illumination under photopic and scotopic conditions to identify the types of photoreceptor input to these nuclei. 2. The majority of visually responsive SCN neurones studied under dark adaptation received rod input (48 of 52, 92 %). The action spectrum conformed to the sensitivity of rhodopsin, with maximal sensitivity at around 505 nm. 3. When also studied under light adaptation, most visually responsive SCN neurones (20 out of 26, 77 %) responded to input from cones. The action spectra conformed to the spectrum of green cone opsin, with a main sensitivity peak at 510 nm and a significant secondary peak in the near-ultraviolet region of the spectrum. 4. The frequency of spontaneous activity was typically low under scotopic conditions (range 0.2-17.2 Hz) and higher under photopic conditions (range 0.6-40 Hz) for any given neurone. The most common response under scotopic conditions was an 'on-excitation' (32 of 48, 62.5 %), which changed under photopic conditions to an on-excitation followed by a more prominent off-inhibition. 5. Responses also changed due to endogenous ultradian cycles. Depending on the phase, responses could be altogether absent and even reverted from excitation to inhibition on opposite phases of a cycle. Ultradian cycles had a circadian dependence and were most common at around the light phase:dark phase (L:D) and D:L transition points of the circadian cycle. 6. Under photopic conditions, SCN neurones showed rhythmic electrical activity, with a preferred firing interval that had a value between 18 and 39 ms. This rhythmic activity was probably the result of endogenous subthreshold membrane potential oscillations. 7. In conclusion, light acting either via rod or cone pathways could have powerful, opposing actions on SCN neurones. These actions were state dependent. The presence of these neuronal responses suggests a role for rod and cone photoreceptors in SCN function.
The scotopic ERG recorded in response to a focal stimulus has a double b-wave. The first wave results from direct focal stimulation of the retina, and the second originates in the peripheral retina in response to scattered light. The aim of this study was to assess two possible protocols for the isolation of the focal rod response, namely 'Subtraction' and 'Background Adaptation' techniques. The Subtraction technique involved the recording of a full-field response, which matched the b-wave elicited by intraocular light scatter, and the subtraction of this full-field response from the initial trace to isolate the focal component. In the Background Adaptation technique an adapting surround was used to suppress the response from the peripheral retina. Focal rod responses were isolated with both techniques. However, the Background Adaptation technique was found to more reliably elicit a focal response with a measurable a-wave, and was also considerably less time consuming than the Subtraction technique.
The spatial and temporal summation of light by the receptive field centre of frog retinal ganglion cells were studied by extracellular recording in the eyecup preparation. The purpose was to quantify how summation changes with the state of light and dark adaptation and to clarify whether changes are due to the transition between rod and cone vision. Spatial summation was found to decrease by 30-50% as the cell was light-adapted to a threshold some 4 log units above the dark-adapted one. Temporal summation for threshold responses fell as the power -0.17 of the intensity of an adapting steady background. Neither change was bound to the rod-cone transition but occurred in the ranges of both receptor types; at equal sensitivities the summation of both receptor systems was matched.
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The spatial characteristics of the oscillatory potentials (OPs) and the a- and b-waves of the frog electroretinogram (ERG) were studied during different states of adaptation induced by repetitive light stimulus given at 1 min or 15 sec interval. The spatial integration area of the OPs was found to be reduced during relatively more light-adapted conditions compared to relatively more dark-adapted ones. No major adaptive effect on the spatial summation area of the a- and b-waves observed. The results indicate that the spatial changes of the OPs observed may be signalled by the amacrine cells which would thus be indirectly involved in the generation of the OPs. The spatial properties of the OPs differed from that of the a- and b-waves suggesting that the OPs have a different origin to that of the a- and b-waves. Finally, the present data indicate a clear and sensitive spatial organization in the frog ERG.