Michael A. Sikora

Hippocampal Sclerosis in Temporal Lobe Epilepsy: Findings at 7 T

PURPOSE To determine if ultrahigh-field-strength magnetic resonance (MR) imaging can be used to detect subregional hippocampal alterations. MATERIALS AND METHODS Subjects provided written consent to participate in this prospective institutional review board-approved HIPAA-compliant study. T1- and T2-weighted 7-T brain MR images were acquired in 11 healthy subjects and eight patients with temporal lobe epilepsy (TLE). In all subjects, images were qualitatively examined for evidence of hippocampal atrophy, signal change, and malrotation with the Bernasconi definition, and digitations of the hippocampal heads were counted (agreement was measured with the κ statistic). Data were analyzed quantitatively with manual subregional hippocampal body segmentation. Subregional data in individual subjects with TLE were compared with data in control subjects to detect deviation from the control range for volume measures on each side and with asymmetry indexes. RESULTS All eight patients with TLE had hippocampal abnormalities on the epileptogenic side. Subregional analysis revealed selective lateral Ammon horn atrophy in six patients and diffuse Ammon horn and dentate gyrus atrophy in one patient. Paucity of hippocampal digitations occurred on the epileptogenic side in all patients with TLE and also on the contralateral side in three patients (interrater κ value, 0.80). Hippocampal malrotation was observed in three patients with TLE and four control subjects. CONCLUSION Ultrahigh-field-strength MR imaging permitted detection of selectively greater Ammon horn atrophy in patients with TLE and hippocampal sclerosis. Paucity of digitations is a deformity of the hippocampal head that was detected independent of hippocampal atrophy in patients with mesial TLE.

Responses of ganglion cells to contrast steps in the light-adapted retina of the tiger salamander

The impulse discharge of single ganglion cells was recorded extracellularly in superfused eyecup preparations of the tiger salamander (Ambystoma tigrinum). Contrast flashes (500 ms) were applied at the center of the receptive field while the retina was light adapted to a background field of 20 cd/m2. The incidence of cell types in a sample of 387 cells was: ON cells (4%), OFF cells (28%), and ON/OFF cells (68%). Quantitative contrast/response measurements were obtained for 83 cells. On the basis of C50, the contrast necessary to evoke a half-maximal response, ON/OFF cells fell into 3 groups: (1) Positive Dominant (26%), (2) Balanced (23%), and (3) Negative Dominant (51%). Positive Dominant cells tended to be relatively contrast insensitive. On the other hand, many Negative Dominant cells showed remarkably low C50 values and very steep contrast/response curves. Contrast gain to negative contrast averaged 8.5 impulses/s/% contrast, some four times greater than that evoked by positive contrast. In most ON/OFF cells, the latency of the first spike evoked by a negative contrast step was much shorter (40-100 ms) than that evoked by a positive contrast step of equal contrast. OFF cells typically showed higher C50 values, larger dynamic ranges, and longer latencies than those of Negative Dominant ON/OFF cells. Thus, different pathways or mechanism apparently mediate the off responses of OFF and ON/OFF cells. In sum, the light-adapted retina of the tiger salamander is strongly biased in favor of negative contrast, as shown by the remarkably high contrast sensitivity and faster response of Negative Dominant cells, the remarkably low incidence of ON cells, and the insensitivity of Positive Dominant cells. Some possible underlying influences of bipolar and amacrine cells are discussed.

Neurophysiologic Assessment of Endolymphatic Hydrops

An electrophysiological method for assessing endolymphatic hydrops of the cochlea in an animal model of Menieres disease is described. Eighth nerve gross action potentials (AP) were evoked by tone bursts presented at discrete phases of a simultaneously delivered 50-Hz pure tone. Hydropic cochleas showed significantly less modulation of AP amplitude and latency than normal cochleas. This method may be useful in objectively evaluating Menieres disease with electrocochleography.

Ototoxicity of ethanol in the tympanic cleft in animals.

This work was undertaken to study the ototoxicity of topically applied ethanol in quantitative terms. Using guinea pigs, ethanol was administered (1) on the round window for 10 min, (2) instilled in the middle ear cavity for 24 hours, and (3) perfused into the cochlear canal (into the scala tympani) at the rat of 10 microliter/min for 10 min. Cochlear microphonics from the electrode on the round window were recorded. The critical concentration (the maximum dilution ratio) of the ethanol that appears to be ototoxic in the experimental condition mentioned above was determined to be (1) 50%, (2) 10%, (3) 0.1% respectively. Using chinchillas, the effect of ethanol with round window application on the Endocochlear Potential (EP) was studied. Simultaneous recording of EP from the 1st and 3rd turn of the cochlea showed a more marked decline in EP in the 1st turn. 70% ethanol caused an irreversible, plateauing decline in EPO, while 35% ethanol caused a reversible decline in EP.

Measurement of action potential thresholds in experimental endolymphatic hydrops

Unilateral experimental endolymphatic hydrops was created by endolymphatic sac and duct obstruction in guinea pigs. Studies of action potential (AP) threshold were then performed and compared with histologic findings. Action potential thresholds were significantly increased in the hydrops ears. The close relationship between AP changes and hydrops was sufficient to allow the detection of hydrops by AP threshold measurements alone. The AP threshold changes were positively related to the degree of hydrops and postoperative interval in a manner consistent with hearing changes seen in Menieres disease. Based upon the foregoing, experimental endolymphatic hydrops appears to be a valid and usable model for the study of the cochlear aspects of Menieres disease.

Diet-induced Hyperlipidemia and Auditory Dysfunction

Chinchillas rendered hyperlipidemic by a 1% cholesterol diet or maintained on a normal diet were either exposed to a 2-octave bandpass noise (700-2,800 Hz for 220 min at 105 or 114 dB) or else not exposed to noise. The animals were assessed with tone-burst (2-16 kHz) elicited compound action potentials (CAP). Compared with normal diet animals, the hyperlipidemic animals: not exposed to noise exhibited elevated thresholds at 8 kHz and higher frequencies; exposed to 105-dB noise exhibited elevated thresholds at 16 kHz; and exposed to 114-dB noise exhibited elevated thresholds at 2-16 kH. Surface preparations were made of the left cochleae of all noise-exposed animals. There was essentially no difference in hair cell counts between hyperlipidemic animals exposed to the 105-dB noise and normal animals similarly exposed. The hyperlipidemic animals exposed to the 114-dB noise exhibited a greater hair cell loss in the first turn of the cochlea than did similarly exposed normal animals. We conclude that maintenance on a high-cholesterol diet can cause a high-frequency hearing loss, probably due to vascular pathology resulting from a hyperlipidemic state. Furthermore, maintenance on a high-cholesterol diet can increase susceptibility to noise-induced hearing losses.

A computational model of the ribbon synapse.

A model of the ribbon synapse was developed to replicate both pre- and postsynaptic functions of this glutamatergic juncture. The presynaptic portion of the model is rich in anatomical and physiological detail and includes multiple release sites for each ribbon based on anatomical studies of presynaptic terminals, presynaptic voltage at the terminal, the activation of voltage-gated calcium channels and a calcium-dependent release mechanism whose rate varies as a function of the calcium concentration that is monitored at two different sites which control both an ultrafast, docked pool of vesicles and a release ready pool of tethered vesicles. The postsynaptic portion of the program models diffusion of glutamate and the physiological properties of glutamatergic neurotransmission in target cells. We demonstrate the behavior of the model using the retinal bipolar cell to ganglion cell ribbon synapse. The model was constrained by the anatomy of salamander bipolar terminals based on the ultrastructure of these synapses and presynaptic contacts were placed onto realistic ganglion cell morphology activated by a range of ribbon synapses (46-138). These inputs could excite the cell in a manner consistent with physiological observations. This model is a comprehensive, first-generation attempt to assemble our present understanding of the ribbon synapse into a domain that permits testing our understanding of this important structure. We believe that with minor modifications of this model, it can be fine tuned for other ribbon synapses.

Experimental hypercholesterolemia and auditory function in the chinchilla.

Possible harmful effects of a high-cholesterol diet on auditory function were suggested by our previous work in rabbits, in which evoked potentials were measured from a chronic electrode inserted into the inferior colliculus. However, serum cholesterol levels in those rabbits tended to be extraordinarily high, ie, more than 1,500 mg/dL. Chinchillas were used in the present work as an animal model to study the relationship between hypercholesterolemia and auditory dysfunction. One percent cholesterol in standard Chinchow was fed to chinchillas for three months. The experimental groups showed a high mean cholesterol level of 437 ± 394 mg/dL (N = 9). Isopotential curve of the cochlear microphonics, threshold of action potentials (AP), and endocochlear DC potential did not differ from those in the control group. When moderately intense sound (12 kHz, 95 dB SPL) was given for ten minutes, however, the reduction in AP threshold was significantly greater (P=.036) in the cholesterol group. It is postulated that hypercholesterolemia may be one of the factors involved in differential susceptibility to noise.

Structure and functional connections of presynaptic terminals in the vertebrate retina revealed by activity-dependent dyes and confocal microscopy

The fluorescent dyes sulforhodamine 101 (SR 101) and FM1‐43 were used as activity‐dependent dyes (ADDs) to label presynaptic terminals in the retinas of a broad range of animals, including amphibians, mammals, fish, and turtles. The pattern of dye uptake was studied in live retinal preparations by using brightfield, fluorescence, and confocal microscopy. When bath‐applied to the retina‐eyecup, these dyes were avidly sequestered by the presynaptic terminals of virtually all rods, cones, and bipolar and amacrine cells; ganglion cell dendrites and horizontal cells lacked significant dye accumulation. Other structures stained with these dyes included pigment epithelial cells, cone outer segments, and Müller cell end‐feet. Studies of dye uptake in dark‐ and light‐adapted preparations showed significant differences in the dye accumulation pattern in the inner plexiform layer (IPL), suggesting a dynamic, light‐modulated control of endocytotic activity. Presynaptic terminals in the IPL could be segregated on the basis of volume: bipolar varicosities in the IPL were typically larger than those of amacrine cells. The combination of retrograde labeling of ganglion cells and presynaptic terminal labeling with ADDs served as the experimental preparation for three‐dimensional reconstruction of both structures, based on dual detector, confocal microscopy. Our results demonstrate a new approach for studying synaptic interactions in retinal function. These findings provide new insights into the likely number and position of functional connections from amacrine and bipolar cell terminals onto ganglion cell dendrites. J. Comp. Neurol. 437:129–155, 2001.

Natural images and contrast encoding in bipolar cells in the retina of the land- and aquatic-phase tiger salamander

Intracellular recordings were obtained from 57 cone-driven bipolar cells in the light-adapted retina of the land-phase (adult) tiger salamander (Ambystoma tigrinum). Responses to flashes of negative and positive contrast for centered spots of optimum spatial dimensions were analyzed as a function of contrast magnitude. On average, the contrast/response curves of depolarizing and hyperpolarizing bipolar cells in the land-phase animals were remarkably similar to those of aquatic-phase animals. Thus, the primary retinal mechanisms mediating contrast coding in the outer retina are conserved as the salamander evolves from the aquatic to the land phase. To evaluate contrast encoding in the context of natural environments, the distribution of contrasts in natural images was measured for 65 scenes. The results, in general agreement with other reports, show that the vast majority of contrasts in nature are very small. The efficient coding hypothesis of Laughlin was examined by comparing the average contrast/response curves of bipolar cells with the cumulative probability distribution of contrasts in natural images. Efficient coding was found at 20 cd/m2 but at lower levels of light adaptation, the contrast/response curves were much too shallow. Further experiments show that two fundamental physiological factors-light adaptation and the nonlinear transfer across the cone-bipolar synapse are essential for the emergence of efficient contrast coding. For both land- and aquatic-based animals, the extent and symmetry of the dynamic range of the contrast/response curves of both classes of bipolar cells varied greatly from cell to cell. This apparent substrate for distributed encoding is established at the bipolar cell level, since it is not found in cones. As a result, the dynamic range of the bipolar cell population brackets the distribution of contrasts found in natural images.