Dark adaptation is faster in pigmented than albino rats

Purpose: Previous reports have raised the possibility that, compared to pigmented rats, albino rats might be night blind. The purpose of this study was to reinvestigate this issue by comparing the dark-adaptation process of the pigmented Long-Evans (LE) and albino Sprague Dawley (SD) rats. Methods: Scotopic ERGs obtained from LE and SD rats were recorded following periods of dark adaptation 0.5, 3 and 12 h. Intensity response functions were generated with flashes of white light spanning over a 7 log-unit range with a maximal intensity of 8 cd.s.m^–2 in energy. Results: SD rats showed a gradual increase in the amplitude of the scotopic b-wave Vmax (maximal `saturated' rod b-wave amplitude) and retinal sensitivity (k) as the duration of the dark-adaptation period increased. In contrast, LE rats did not demonstrate any further significant gain in retinal function (Vmax or k) beyond 30 min of dark-adaptation. Thus for periods of dark-adaptation of 30 min or less, the rod function of the LE rats is superior to that of the SD rats while both strains have comparable retinal functions following 3 h or more of dark-adaptation. Conclusions: Our results indicate that LE and SD rats differ in their rapidity to dark-adapt, a finding that could explain the previous claim that SD rats were night blind. The reduced bio-availability of calcium ions in eyes lacking melanin could explain this difference. Calcium was previously shown to play a key role in retinal adaptation processes.     

Tapetum lucidum in the pigmented and albino ferret

Light and electron microscopy showed that the tapetum lucidum in the pigmented ferret is morphologically indistinguishable from that in the albino ferret. The matrix of the rods of the tapetal cells was strongly osmiophilic, but glutaraldehyde fixation before osmium tetroxide treatment caused a dissolution of the matrix material. It has been proposed that the tapetal cells are modified melanocytes and that the tapetal rods are composed of melanin, but it can be concluded from our data that the matrix of the tapetal rods is not melanin. Further studies by plasma-atomic emission spectrometry showed that the tapetal cells are very rich in zinc, with similar levels in pigmented and albino ferrets. Excessive concentrations of other metals were not observed. Histochemical demonstration of heavy metal showed that the zinc is present in the tapetal rods and indicated a localization mainly in the rod membranes.     

ERG changes in albino and pigmented mice after optic nerve transection

Optic nerve transection (ONT) triggers retinal ganglion cell (RGC) death. By using this paradigm, we have analyzed for the first time in adult albino and pigmented mice, the effects of ONT in the scotopic threshold response (STR) components (negative and positive) of the full-field electroretinogram. Two weeks after ONT, when in pigmented mice approximately 18% of the RGC population survive, the STR-implicit time decreased and the p and nSTR waves diminished approximately to 40% or 55%, in albino or pigmented, respectively, with respect to the values recorded from the non-operated contralateral eyes. These changes were maintained up to 12 weeks post-ONT, demonstrating that the ERG-STR is a useful parameter to monitor RGC functionality in adult mice.     

Effects of APB,PDA, and TTX on ERG responses recorded using both multifocal and conventional methods in monkey. Effects of APB,PDA, and TTX on monkey ERG responses

Results from studies of human subjects suggest that the multifocal ERG technique developed by Erich Sutter and colleagues has considerable potential for assessment of retinal function in both the clinic and laboratory. While the utility of this measure depends to a large extent upon an understanding of the physiological origin for the different response components, relatively little is known in this regard. For the experiments described in this report, we made ERG recordings using both multifocal and conventional methods. Intravitreal injections of APB, PDA, and TTX were used to identify contributions from activity in ON pathway, OFF pathway, and third order retinal neurons, respectively. The results show that photoreceptor activity makes a small direct contribution to 1st and 2nd order multifocal photopic luminance responses. TTX-sensitive activity in third order retinal neurons contributes to both 1st and 2nd order responses with relatively greater contribution to the 2nd order response. Blockade of TTX-sensitive activity in third order cells produces effects on the 2nd order response which are very similar to changes observed in eyes suffering selective loss of retinal ganglion cells resulting from experimental glaucoma. Effects of these intravitreally injected test agents were also determined, in the same recording session, for flash, 30 Hz flicker, and oscillatory potential responses.     

Development of postreceptoral function in pigmented and albino guinea pigs

Retinal neurons are generated in overlapping growth spurts with ganglion cell and cone populations peaking sooner than rod and bipolar cell numbers. As such the functional development of the inner and outer retinal components and elements within these strata (rods vs. cones) may differ. We considered the postnatal development of the postreceptoral components of the ERG (P2, oscillatory potentials) in the guinea pig. ERGs were also evaluated across albino and pigmented strains in order to consider the role that pigmentation has for functional development. Electroretinograms were collected on postnatal days PDI to PD60 (n = 4-7 per time point). The postreceptoral P2 amplitude and implicit time was extracted (digital subtraction of modelled P3 and filtering, 0.5-49 Hz). Intensity-response relationships were described using Naka-Rushton functions whose parameters were compared using a nonparametric bootstrap. Oscillatory potentials (OPs) were extracted following signal conditioning and filtering to remove the a- and b-waves and were described using a Gabor function. OP response parameters were compared using repeated measures ANOVA. Postreceptoral P2 amplitudes mature soon after birth (PD10-PD12). Oscillatory potentials show a similar postnatal amplitude development (PD10-PD12) but a later maturation in timing (PD20) compared with the postreceptoral waveform. All components (P3, P2, and OPs) declined at the same relative rate with age after PD12. Albino animals gave larger, faster, and more sensitive waveforms at all ages but showed the same age-related trends as did pigmented animals. Early development of inner retinal synapses in guinea pigs may underlie the rapid postnatal maturation of their postreceptoral response. These appear to be constrained by the development of receptoral responses. All components declined at the same rate suggesting either a change in the photoreceptoral response or changes to ocular impedance with age.     

The ERG of guinea pig (Cavis porcellus): comparison with I-type monkey and E-type rat

We have been in search of an alternate species for the monkey to study the effects of drugs on the I-type photopic electroretinogram (ERG) response that is typically seen in the cone-rich retina of the primate. The guinea pig has two types of cones, one of which contains a middle-wavelength sensitive pigment otherwise found only in Old World primates. We studied the Ganzfeld electroretinogram (ERG) of the guinea pig in relation to monkey and rat ERGs to learn whether the guinea pig might be a good animal model to study the `primate-like' cone ERG. The guinea pig scotopic ERG was similar to other mammal ERGs and was not electronegative when fully dark-adapted. We saw no evidence of a negative-going scotopic threshold response (STR). The guinea pig photopic ERG a-wave is larger than that of the rat but much smaller than the primate a-wave, and it lacked a phasic d-wave. PDA eliminated guinea pig photopic a-wave and caused the OFF-response to long stimuli to invert polarity, as seen in monkey but not in rat. The guinea pig overall shows a weak I-type response and may be a useful substitute for primate in some studies of the photopic ERG.     

球内铅异物伤时ERG与视网膜形态变化

研究了兔眼球内铅异物伤后的视网膜电图(ERG)和视网膜形态学变化。伤后2个月,ERG a 波降低75%,b 波降低66%;伤后3个月,视网膜各层细胞广泛出现肿胀、变性、甚至坏死。并对实验结果进行了分析和讨论。提出球内铅异物伤后,应在4周内手术取出异物。     

Two distinct processes are evident in rat cone flicker ERG responses at low and high temporal frequencies

The electroretinogram (ERG) provides a noninvasive, objective measure of retinal function, and is one of the most widely used diagnostic tools in the study of visual disorders. Although rodents are often used in the study of retinal disease, the properties of the flicker ERG of the rodent retina have not been fully characterized. Here, we show that the fundamental response of the rat ERG to sine-wave flicker exhibited a low-pass pattern in the frequency range from 2 to 30 Hz, whereas the second harmonic (F2) showed a more complex frequency-response relation. The F2 component represented only a small fraction of the ERG response at low temporal frequencies (below 12 Hz), but it made a substantial contribution to responses at high frequencies. The contrast-response relation was linear when tested with a low-frequency (6 Hz) stimulus, but saturated in response to a high-frequency (20 Hz) stimulus. After intravitreal injection of L-AP4, a specific blocker of the retinal ON pathway, the flicker responses elicited by either 6- or 20-Hz stimuli were greatly reduced in amplitude, whereas only a very slight enhancement was seen after the application of PDA, a drug that blocks retinal OFF-pathway activity. Based on the observed differences in the degree of nonlinearity, and contrast-response properties of the rat flicker ERG at low and high frequencies, as well as the pharmacological results, we postulate that sustained and transient ON bipolar cells generate the flicker ERG responses elicited at low and high temporal frequencies, respectively.     

A comparison of oscillatory potential and pattern electroretinogram measures in diabetic retinopathy

Both basic and clinical electrophysiological investigations have established that the oscillatory potentials (OP) and pattern electroretinogram (PERG) appear to originate from retinal sites that are in proximity. The OPs, subcomponents of the flash ERG, have been shown to reflect disturbances in retinal circulation, and OP amplitude attenuation or loss may be a distinctive feature of diabetic retinopathy. The PERG has been shown to be abnormal in diseases of the optic nerve and ganglion cell body. Thus its relative sensitivity for detection of electroretinal abnormalities in diabetic retinopathy is in question. This study assessed the sensitivity of ERG and OP measures in their detection of abnormalities of electroretinal function in diabetic patients referred to our laboratory. Thirty-five adult Type I patients were studied: 21 with background retinopathy (BR group), 14 with no evidence of background retinopathy (No BR group), and 25 normal control subjects.Monocular OPs were recorded to full-field ganzfeld stimulation at four stimulus intensities. PERGs were obtained from checkerboard pattern reversal stimulation (checksize = 30 arc). Peak-to-peak amplitude and peak implicit time measures of PERGs and OPs were obtained. Subsequent multivariate analysis demonstrated significant differences between normals and diabetic patients, including diabetics with no clinical evidence of retinopathy. In addition, the OP and PERG implicit times appear to be unaffected while OP and PERG amplitudes were diminished in patients with background retinopathy. Only OP amplitudes were found to be significantly diminished in diabetic patients with no photographic evidence of background retinopathy. The PERGs were normal in these patients. Overall, the OP amplitude measures were more sensitive than PERG measures in detecting abnormalities in patients with no retinal photographic evidence of background retinopathy.     

Development of ERG responses: The ISCEV rod, maximal and cone responses in normal subjects

PURPOSE: Summarize ISCEV ERG responses from normal infants and children. METHODS: The amplitudes and implicit times of the ISCEV rod, maximal dark-adapted and cone responses from a total of 409 normal infants (n = 128), children and adult controls were compiled. The subjects, aged 1 week to 52 years, were divided into seven age groups, including four in infancy (< 52 weeks). The response parameters for each age group were summarized as percentiles. RESULTS: In each ISCEV condition, the youngest infants (1-5 weeks) had significantly smaller amplitudes and longer implicit times than adults. Amplitude increased and implicit time decreased systematically with age. CONCLUSIONS: The developmental changes in ERG responses are significant. The medians and ranges herein provide provisional norms against which the ERG responses from pediatric patients can be compared.     

Asymmetry of focal macular photopic negative responses (PhNRs) in monkeys

The photopic negative response (PhNR) is a slow, negative-going wave of the photopic electroretinogram (ERG) that appears after the b-wave. Recent studies have shown that the PhNR originates from the spiking activities of inner retinal neurons including the ganglion cells and their axons. The aim of this study was to determine whether there is any asymmetry in the amplitude of the PhNR elicited from the upper and lower macular areas, and between the nasal and temporal macular areas in rhesus monkeys. To accomplish this, we recorded focal macular PhNRs that were elicited by red hemi-circular stimuli presented on a blue background. We show that the PhNR from the upper macular area was significantly larger than that of the lower macular area, and the PhNR of the nasal macula was significantly larger than that of the temporal macula. These asymmetries were present in the focal PhNR elicited by both brief and long duration stimuli, and the asymmetries were completely eliminated by an intravitreal injection of tetrodotoxin (TTX). These results suggest that the upper-lower and nasal-temporal asymmetries of PhNR in the primate retina are mainly caused by TTX-sensitive spiking activities of inner retinal neurons.     

Investigating structural and biochemical correlates of ganglion cell dysfunction in streptozotocin-induced diabetic rats

The aim of this study was to determine whether inner retinal dysfunction in diabetic rats is correlated with structural and/or biochemical changes in the retina and optic nerve. Using the electroretinogram (ERG; −5.83 to 1.28 log cd.s.m⁻²) retinal function (photoreceptor, bipolar, amacrine and ganglion cell components) was measured in control (n = 13; citrate buffer) and diabetic (n = 13; streptozotocin, STZ, 50 mg kg⁻¹) rats, 12 weeks following treatment. Retinae and optic nerves were analyzed for structural changes and retinae were assessed for alterations in growth factor/cytokine expression using quantitative real-time PCR. We found that phototransduction efficiency was reduced 12 weeks after STZ-induced diabetes (−30%), leading to reduced amplitude of ON-bipolar (−18%) and amacrine cell (−29%) dominated responses; ganglion cell dysfunction (−84%) was more profound. In the optic nerve, nerve fascicle area and myelin sheath thickness were reduced (p < 0.05), whereas the ratio of blood vessels and connective tissue to total nerve cross-sectional area was increased (p < 0.05) in diabetic compared to control rats. In the retina, connective tissue growth factor (CTGF), transforming growth factor beta, type 2 receptor (TGFβ-r2) mRNA and platelet-derived growth factor B (PDGF-B) mRNA were increased (p < 0.035). Reduced ganglion cell function was correlated with increased CTGF and TGFβ-r2, but not PDGF-B mRNA. In summary, the ganglion cell component exhibited the greatest level of dysfunction within the ERG components examined after 12 weeks of STZ-induced diabetes; the level correlated with increased CTGF and TGFβ-r2 mRNA, but not with gross morphological changes in the retina or optic nerve.     

Electrophysiological findings in a family with congenital arteriohepatic dysplasia (Alagille syndrome)

Arteriohepatic dysplasia (Alagille syndrome) is a congenital cholestatic disease associated with ocular abnormalities. Three Japanese siblings, a 14-year-old girl, an 11 year-old boy, and a 9-year-old girl with this syndrome were studied. All three patients showed neonatal jaundice, hepatic dysfunction, characteristic facies, and psychomental retardation. The two sisters had cardiac murmurs. Ophthalmological examinations revealed that they had posterior embryotoxon, refractive error, retinochoroidal degener ation, and electrophysiological abnormalities. The two sisters showed retinochoroidal degeneration and unilateral high myopia while the brother showed marked retino choroidal degeneration with extensive pigment clumps. Visual fields showed moderate concentric contraction in the two sisters and marked concentric contraction in the brother. Amplitudes of the single flash electroretinogram were moderately reduced in the sisters, the test was nonrecordable in one eye and extensively reduced in the other eye of the brother. The electrooculogram was borderline in the elder sister and abnormal in the brother and younger sister. Visual evoked cortical potential (VECP) were abnormal in the high myopic eye in each of the two sisters. Ophthalmological findings including electrophysiological examinations may help to confirm the diagnosis of this multisystem familial disorder.     

Neuronal ceroid lipofuscinosis in the Polish Owczarek Nizinny (PON) dog

Visual dysfunction and neurological symptoms were found in Polish Owczarek Nizinny (PON) dogs. Two dogs were examined, one at 2 years of age and the other one at 4 years. The oldest dog was totally blind. The 2-year-old dog developed mental disturbances and the 4-year-old dog became severely ataxic. Ophthalmoscopical findings were retinal hyper-reflectivity, attenuation of the retinal vessels and the presence of greyish to brown spots in the fundus. Electrophysiological and ultrastructural studies were performed in the 2-year-old dog. Scotopic ERG responses were absent, whereas 30 Hz cone flicker responses were recordable, although with an amplitude reduced to about 30% of the normal level. A slow negative potential replaced the c-wave, indicating a dysfunction of the RPE. Intracellular inclusions with a granular appearance or containing membranous fingerprint-like or curvilinear profiles, resembling ceroid, were found in different retinal cells. The RPE cells in the central areas were charged with autofluorescent material having similar structure, Photoreceptor degeneration was most severe in the central areas, corresponding to the RPE changes. It appears than the PON dog may provide a new animal model for neuronal ceroid lipofuscinosis.     

Cataract and Glaucoma Development in Juvenile Neuronal Ceroid Lipofuscinosis (Batten Disease)

Background: Ophthalmologic studies of Juvenile Neuronal Ceroid Lipofuscinosis (JNCL) have focused mainly on retinal involvement, and so far no anterior segment abnormalities have been described. In the present study, we report the findings of pre-senile cataract in five patients with JNCL.

Material and Methods: Our sample consisted of 35 patients (19 males, 16 females) with JNCL associated to the Centre for Rare Disease, Aarhus University Hospital. They represent all patients with JNCL born in Denmark in the period 1971–2003. At the half-yearly routine outpatient visits, the anterior section was examined by ordinary penlight without instillation of a mydriatic, and if abnormalities such as cataracts were detected or suspected, the patients were referred for an ophthalmologic examination including slit lamp examination. Follow up was obtained on all patients referred for ophthalmologic examination.

Results: During the study period (1996–2012), five patients were identified with cataract. The patients’ average age at detection of cataract was 20.1?+?1.6 years (mean?+?2SD). Two of the five patients developed acute glaucoma, and in one case prophylactic cataract surgery was performed.

Conclusions: Cataract formation and a secondary acute glaucoma are complications in JNCL which do occur. We recommend that a complete ophthalmological examination of the anterior segment should be performed routinely in patients with JNCL beyond the age of 16 years of age in order to prevent a painful and harmful acute glaucoma which may occur due to mature cataract formation.