Human S-cone electroretinograms obtained by silent substitution stimulation

YesWe used triple silent substitution stimuli to characterize human S-cone electroretinograms (ERGs) in normal trichromats. Short-wavelength-cone (S-cone) ERGs were found to have different morphological features and temporal frequency response characteristics compared to ERGs derived from L-cones, M-cones, and rod photoreceptors in normal participants. Furthermore, in two cases of retinal pathology, blue cone monochromatism (BCM) and enhanced S-cone syndrome (ESCS), S-cone ERGs elicited by our stimuli were preserved and enhanced, respectively. The results from both normal and pathological retinae demonstrate that triple silent substitution stimuli can be used to generate ERGs that provide an assay of human S-cone function.Deutsche Forschungsgemeinschaft (DFG) (KR1317/13-1); Bundesministerium für Bildung und Forschung (BMBF) (01DN14009

The Isolation of Human Rod and Cone Photoreceptor Activity combining Electroretinography and Silent Substitution Techniques

Aims: The electroretinogram (ERG) can be used to independently assess the function of rod and cone photoreceptors within the human retina. The work in this thesis sought to investigate an alternative method of recording the ERG, using the silent substitution paradigm (Estevez and Spekreijse 1982). The aims are separated into two parts, firstly, the isolation and characterisation of the non-dark adapted rod photoreceptor response, and secondly, characterising the ERG response from L-, M- and S-cones. Methods: Rod, L-, M- and S-cone isolating as well as non-isolating sinusoidal flicker and transient square-wave stimuli were generated on a 4 primary LED ganzfeld stimulator to elicit ERGs from non-dark adapted participants with normal and compromised rod or cone function. Results: The results from the rod experiments showed that ERGs elicited by rod isolating silent substitution stimuli exhibit low-pass temporal frequency response characteristics with an upper response limit of 30Hz and saturate beyond 1000ph Td. Responses are optimal between 5 – 8 Hz and between 10-100 photopic Td. There is a significant correlation between the response amplitudes obtained with the silent substitution method and current standard clinical protocols. The results from the cone experiments showed that the L-, M- and S-cone stimulation produced ERGs with very different morphologies. L- and M-cone stimulation is of limited use as an objective measure of colour vision deficiency. Conclusion: Silent substitution provides an effective method for the isolation of human rod and cone photoreceptor function in subjects when stimuli are used within appropriate parameter ranges

Rod Electroretinograms Elicited by Silent Substitution Stimuli from the Light-Adapted Human Eye

YesPurpose: To demonstrate that silent substitution stimuli can be used to generate electroretinograms (ERGs) that effectively isolate rod photoreceptor function in humans without the need for dark adaptation, and that this approach constitutes a viable alternative to current clinical standard testing protocols. Methods: Rod-isolating and non-isolating sinusoidal flicker stimuli were generated on a 4 primary light-emitting diode (LED) Ganzfeld stimulator to elicit ERGs from participants with normal and compromised rod function who had not undergone dark-adaptation. Responses were subjected to Fourier analysis, and the amplitude and phase of the fundamental were used to examine temporal frequency and retinal illuminance response characteristics. Results: Electroretinograms elicited by rod-isolating silent substitution stimuli exhibit low-pass temporal frequency response characteristics with an upper response limit of 30 Hz. Responses are optimal between 5 and 8 Hz and between 10 and 100 photopic trolands (Td). There is a significant correlation between the response amplitudes obtained with the silent substitution method and current standard clinical protocols. Analysis of signal-to-noise ratios reveals significant differences between subjects with normal and compromised rod function. Conclusions: Silent substitution provides an effective method for the isolation of human rod photoreceptor function in subjects with normal as well as compromised rod function when stimuli are used within appropriate parameter ranges. Translational Relevance: This method of generating rod-mediated ERGs can be achieved without time-consuming periods of dark adaptation, provides improved isolation of rod- from cone-based activity, and will lead to the development of faster clinical electrophysiologic testing protocols with improved selectivity

CNGB3 mutations cause severe rod dysfunction

YesCongenital achromatopsia or rod monochromatism is a rare autosomal recessive condition defined by a severe loss of cone photoreceptor function in which rods purportedly retain normal or near-to-normal function. This report describes the results of electroretinography in two siblings with CNGB3-associated achromatopsia. Full field light- and dark-adapted electroretinograms (ERGs) were recorded using standard protocols detailed by the International Society for Clinical Electrophysiology of Vision (ISCEV). We also examined rod-mediated ERGs using series of stimuli that varied over a 6 log unit range of retinal illuminances (−1.9–3.5 log scotopic trolands). Dark-adapted ERGs in achromatopsia patients exhibited severely reduced b-wave amplitudes with abnormal b:a ratios (1.3 and 0.6). In comparison, the reduction in a-wave amplitude was less marked. The rod-mediated ERG took on an electronegative appearance at high-stimulus illuminances. Although the defect that causes achromatopsia is primarily in the cone photoreceptors, our results reveal an accompanying disruption of rod function that is more severe than has previously been reported. The differential effects on the b-wave relative to the a-wave points to an inner-retinal locus for the disruption of rod function in these patients

The use of silent substitution in measuring isolated cone- & rod- human electroretinograms. An electrophysiological study of human rod- and cone- photoreceptor activity derived using silent substitution paradigm

After over a decade of its discovery, the Electroretinogram (ERG) still remains the objective tool that is conventionally used in assessment of retinal function in health and disease. Although there is ongoing research in developing ERG- recording techniques, interpretation and clinical applications, there is still a limited understanding on how each photoreceptor class contribute to the ERG waveform and their role and/or susceptibilities in various retinal diseases still remains unclear. Another limitation with currently used conventional testing protocols in a clinical setting is the requirement of an adaptation period which is time-consuming. Furthermore, the ERG responses derived in this manner are recorded under different stimulus conditions, thus, making comparison of these signals difficult. To address these issues and develop a new testing method, we employed silent substitution paradigm in obtaining cone- and rod- isolating ERGs using sine- and square- wave temporal profiles. The ERGs achieved in this manner were shown to be photoreceptor-selective. Furthermore, these responses did not only provide the functional index of photoreceptors but their contributions to their successive postreceptoral pathways. We believe that the substitution stimuli used in this thesis could be a valuable tool in functional assessment of individual photoreceptor classes in normal and pathological conditions. Furthermore, we speculate that this method of cone/rod activity isolation could possibly be used in developing faster and efficient photoreceptor-selective testing protocols without the need of adaptation.Bradford School of Optometry and Vision Science

The use of Silent Substitution in measuring isolated cone- and rod- Human ERGs

After over a decade of its discovery, the Electroretinogram (ERG) still remains the objective tool that is conventionally used in assessment of retinal function in health and disease. Although there is ongoing research in developing ERG recording techniques, interpretation and clinical applications, there is still a limited understanding on how each photoreceptor class contribute to the ERG waveform and their role and/or susceptibilities in various retinal diseases still remains unclear. Another limitation with currently used conventional testing protocols in a clinical setting is the requirement of an adaptation period which is time consuming. Furthermore, the ERG responses derived in this manner are recorded under different stimulus conditions, thus, making comparison of these signals difficult. To address these issues and develop a new testing method, we employed silent substitution paradigm in obtaining cone- and rod- isolating ERGs using sine- and square- wave temporal profiles. The ERGs achieved in this manner were shown to be photoreceptor-selective. Furthermore, these responses did not only provide the functional index of photoreceptors but their contributions to their successive postreceptoral pathways. We believe that the substitution stimuli used in this thesis could be a valuable tool in functional assessment of individual photoreceptor classes in normal and pathological conditions. Furthermore, we speculate that this method of cone/rod activity isolation could possibly be used in developing faster and efficient photoreceptor-selective testing protocols without the need of adaptation.Bradford School of Optometry and Vision Sciences scholarshi