High-Throughput Binocular Pattern Electroretinograms in the Mouse.

The pattern electroretinogram (PERG) reflects the electrical activity of retinal ganglion cells (RGC) and has become the most used technology to assess RGC function in experimental models of glaucoma and optic neuropathies. We describe a novel method for obtaining user-friendly, robust PERG simultaneously from each eye using asynchronous binocular stimulation and one-channel acquisition of signals recorded from a subcutaneous needle in the snout.

Adaptation of retinal ganglion cell function during flickering light in the mouse.

Rapid dilation of retinal vessels in response to flickering light (functional hyperemia) is a well-known autoregulatory response driven by increased neural activity in the inner retina. Little is known about flicker-induced changes of activity of retinal neurons themselves. We non-invasively investigated flicker-induced changes of retinal ganglion cell (RGC) function in common inbred mouse strains using the pattern electroretinogram (PERG), a sensitive measure of RGC function. Flicker was superimposed on the pattern stimulus at frequencies that did not generate measurable flicker-ERG and alter the PERG response. Transition from flicker at 101 Hz (control) to flicker at 11 Hz (test) at constant mean luminance induced a slow reduction of PERG amplitude to a minimum (39% loss in C57BL/6J mice and 52% loss in DBA/2J mice) 4-5 minutes after 11 Hz flicker onset, followed by a slow recovery to baseline over 20 minutes. Results demonstrate that the magnitude and temporal dynamics of RGC response induced by flicker at 11 Hz can be non-invasively assessed with PERG in the mouse. This allows investigating the functional phenotype of different mouse strains as well as pathological changes in glaucoma and optic nerve disease. The non-contact flicker-PERG method opens the possibility of combined assessment of neural and vascular response dynamics.

High-Throughput Binocular Pattern Electroretinograms in the Mouse.

We describe a new method for obtaining user-friendly, robust pattern electroretinograms (PERG) simultaneously from each eye using asynchronous binocular stimulation and one-channel acquisition of signals recorded from a subcutaneous needle in the snout.

Next Generation PERG Method: Expanding the Response Dynamic Range and Capturing Response Adaptation.

PURPOSE: To compare a new method for steady-state pattern electroretinogram (PERGx) with a validated method (PERGLA) in normal controls and in patients with optic neuropathy. METHODS: PERGx and PERGLA were recorded in a mixed population (n = 33, 66 eyes) of younger controls (C1; n = 10, age 38 ± 8.3 years), older controls (C2; n = 11, 57.9 ± 8.09 years), patients with early manifest glaucoma (G; n = 7, 65.7 ±11.6 years), and patients with nonarteritic ischemic optic neuropathy (N; n = 5, mean age 59.4 ± 8.6 years). The PERGx stimulus was a black-white horizontal grating generated on a 14 × 14 cm LED display (1.6 cycles/deg, 15.63 reversals/s, 98% contrast, 800 cd/m2 mean luminance, 25° field). PERGx signal and noise were averaged over 1024 epochs (∼2 minutes) and Fourier analyzed to retrieve amplitude and phase. Partial averages (16 successive samples of 64 epochs each) were also analyzed to quantify progressive changes over recording time (adaptation). RESULTS: PERGLA and PERGx amplitudes and latencies were correlated (Amplitude R2 = 0.59, Latency R2 = 0.39, both P < 0.0001) and were similarly altered in disease. Compared to PERGLA, however, PERGx had shorter (16 ms) latency, higher (1.39×) amplitude, lower (0.37×) noise, and higher (4.2×) signal-to-noise ratio. PERGx displayed marked amplitude adaptation in C1 and C2 groups and no significant adaptation in G and N groups. CONCLUSIONS: The PERGx high signal-to-noise ratio may allow meaningful recording in advanced stages of optic nerve disorders. In addition, it quantifies response adaptation, which may be selectively altered in glaucoma and optic neuropathy. TRANSLATIONAL RELEVANCE: A new PERG method with increased dynamic range allows recording of retinal ganglion cell function in advanced stages of optic nerve disorders. It also quantifies the response decline during the test, an autoregulatory adaptation to metabolic challenge that decreases with age and presence of disease.

Relationship between transient and steady-state pattern electroretinograms: theoretical and experimental assessment.

PURPOSE: We determined if the overlap of transient (tr) pattern electroretinograms (PERG(tr)) can explain the generation of the steady-state (SS) pattern electroretinogram (PERG(SS)), and investigated the relationship between the two types of responses. METHODS: Slightly jittered pattern reversals were used to generate quasi SS (QSS) PERG(SS) responses from eight normal subjects, recorded using lower eyelid skin electrodes, at rates between 6.9 and 26.5 reversals per second (rps). Jittered quasi PERG(SS) were deconvolved using the frequency domain continuous loop averaging deconvolution method. Additionally, conventional PERG(tr) at 2.2 rps and PERG(SS) at each of the QSS stimulation rates were obtained from all subjects. Two synthetic PERG(SS) responses were constructed at each stimulation rate, one using the PERG(tr) obtained at that rate, and the other using the conventional 2.2 rps PERG(tr). Synthetic responses then were compared to the recorded PERG(SS) using amplitude, latency, and spectral measurements. RESULTS: Findings indicate that the PERG(SS) obtained at SS rates can be predicted using the superposition of deconvolved tr PERGs at each particular rate. Although conventional PERG(tr) can explain PERG(SS) obtained at rates below 15.4 rps (≥ 97% correlation), for higher reversal rates only deconvolved responses obtained at that rate can produce the recorded SS responses (96% vs. 65% correlation at 26.5 rps). CONCLUSIONS: The study shows that PERG(SS) results from the overlapping of tr PERG(tr) waveforms generated at that reversal rate. The first two peaks (N(SS) and P(SS)) of the PERG(SS) reflect N35 and P50 waves of the tr PERG(tr). The N95 amplitude is reduced at conventional (16 rps) SS rates, but contributes to the overall PERG(SS) amplitude.

Unwrapping of transient responses from high rate overlapping pattern electroretinograms by deconvolution.

OBJECTIVE: Due to overlapping, temporal information is mostly lost in high rate steady-state pattern electroretinograms (PERGSS). This study develops a deconvolution method and a display/recording system to "unwrap" PERGSS and obtain a transient, "per stimulus" response (PERGtr) regardless of reversal rate. METHODS: Processing and instrumentation, including high temporal resolution display and acquisition were developed for deconvolving PERGs acquired at high rates by slight jittering of reversal onsets at a given mean rate. RESULTS: The system was successfully tested at eight rates from 2.2 to 78.1rps. At medium rates (17.4-41.2rps) recordings with conventional morphology (N35-P50-N95) but earlier peaks and higher amplitudes were extracted up to 40rps. At higher rates, smaller triphasic responses were obtained, exhibiting similar peak latencies, but reversed polarity. Oscillating potentials (OPs) were also recorded at all rates after deconvolution. CONCLUSIONS: Transient PERGs and OPs can be extracted from quasi steady-state PERG recordings obtained at high rates with a deconvolution algorithm using high temporal resolution display and acquisition systems. SIGNIFICANCE: The methodology to extract transient and oscillatory responses from steady-state PERGs could be useful in understanding high rate responses and diagnosis of various retinal diseases by revealing temporal information on waveform components which cannot be normally observed.

Robust mouse pattern electroretinograms derived simultaneously from each eye using a common snout electrode.

PURPOSE: We recorded pattern electroretinograms (PERGs) simultaneously from each eye in mice using binocular stimulation and a common noncorneal electrode. METHODS: The PERG was derived simultaneously from each eye in 71 ketamine/xylazine anesthetized mice (C57BL/6J, 4 months old) from subcutaneous needles (active, snout; reference, back of the head; ground, root of the tail) in response to contrast-reversal of gratings (0.05 cycles/deg, >95% contrast) generated on two custom-made light-emitting diode (LED) tablets alternating at slight different frequencies (OD, 0.984 Hz; OS, 0.992 Hz). Independent PERG signals from each eye were retrieved using one channel continuous acquisition and phase-locking average (OD, 369 epochs of 492 ms; OS, 372 epochs of 496 ms). The PERG was the average of three consecutive repetitions. RESULTS: Binocular snout PERGs had high amplitude (mean, 25.3 µV, SD 6.6) and no measurable interocular cross-talk. Responses were reliable (test-retest variability within-session, 14%, SD 7; between sessions, 25%, SD 9; interocular asymmetry within-session, 9%, SD 7; between sessions, 13%, SD 5). Retinal ganglion cells (RGCs) were the main source of the binocular snout PERG, as optic nerve crush in three mice abolished the signal. CONCLUSIONS: The PERG, a sensitive measure of RGC function, is used increasingly in mouse models of glaucoma and optic nerve disease. Compared to current methods, the binocular snout PERG represents a substantial improvement in terms of simplicity and speed. It also overcomes limitations of corneal electrodes that interfere with invasive procedures of the eye and facilitates experiments based on comparison between the responses of the two eyes.