Effects of acute high intraocular pressure on red-green and blue-yellow cortical color responses in non-human primates.

Glaucoma is a leading cause of irreversible blindness worldwide, and intraocular pressure (IOP) is an established and modifiable risk factor for both chronic and acute glaucoma. The relationship between color vision deficits and chronic glaucoma has been described previously. However, the effects of acute glaucoma or acute primary angle closure, which has high prevalence in China, on color vision remains unclear. To address the above question, red-green or blue-yellow color responses in V1, V2, and V4 of seven rhesus macaques were monitored using intrinsic-signal optical imaging while monocular anterior chamber perfusions were performed to reversibly elevate IOP acutely over a clinically observed range of 30 to 90 mmHg. We found that the cortical population responses to both red-green and blue-yellow grating stimuli, systematically decreased as IOP increased from 30 to 90 mmHg. Although a similar decrement in magnitude was noted in V1, V2, and V4, blue-yellow responses were consistently more impaired than red-green responses at all levels of acute IOP elevation and in all monitored visual areas. This physiological study in non-human primates demonstrates that acute IOP elevations substantially depress the ability of the visual cortex to register color information. This effect is more severe for blue-yellow responses than for red-green responses, suggesting selective impairment of the koniocellular pathways compared with the parvocellular pathways. Together, we infer that blue-yellow color vision might be the most vulnerable visual function in acute glaucoma patients.

Transduction catalysis: Doxorubicin amplifies rAAV-mediated gene expression in the cortex of higher-order vertebrates.

Rapid and efficient gene transduction via recombinant adeno-associated viruses (rAAVs) is highly desirable across many basic and clinical research domains. Here, we report that vector co-infusion with doxorubicin, a clinical anti-cancer drug, markedly enhanced rAAV-mediated transgene expression in the cerebral cortex across mammalian species (cat, mouse, and macaque), acting throughout the time period examined and detectable at just three days after transfection. This enhancement showed serotype generality, being common to all rAAV serotypes tested (2, 8, 9, and PHP.eB) and was observed both locally and at remote locations consistent with doxorubicin undergoing retrograde axonal transport. All these effects were observed at doses matching human blood plasma levels in clinical therapy and lacked detectable cytotoxicity as assessed by cell morphology, activity, apoptosis, and behavioral testing. Altogether, this study identifies an effective means to improve the capability and scope of in vivo rAAV applications, amplifying cell transduction at doxorubicin concentrations paralleling medical practice.

Antitumor Effects of the Novel Quinazolinone Holu-12: Induction of Mitotic Arrest and Apoptosis in Human Oral Squamous Cell Carcinoma CAL27 Cells.

BACKGROUND/AIM: Quinazolinone is a privileged chemical structure employed for targeting various types of cancer. This study aimed to demonstrate the antitumor activity of synthesized 6,7-disubstituted-2-(3-fluorophenyl) quinazolines (HoLu-11 to HoLu-14). MATERIALS AND METHODS: The cytotoxicity was assessed by the sulforhodamine B (SRB) assay. The cell cycle was examined by flow cytometry. The expression levels of cell cycle- and apoptosis-related proteins were estimated by western blotting. A xenograft animal model was used to explore the antitumor effects of HoLu-12. RESULTS: Among four synthetic quinazolinone derivatives, HoLu-12 significantly reduced the viability of oral squamous cell carcinoma (OSCC) cells. HoLu-12 induced G2/M arrest and increased the expression of cyclin B, histone H3 (Ser10) phosphorylation, and cleaved PARP, indicating that HoLu-12 could induce mitotic arrest and then apoptosis. Moreover, the combination of HoLu-12 and 5-fluorouracil (5-FU) displayed synergistic toxic effect on OSCC cells. HoLu-12 significantly inhibited tumor growth in vivo. CONCLUSION: HoLu-12 induces mitotic arrest and leads to apoptosis of OSCC cells. Furthermore, HoLu-12 alone or in combination with 5-FU is a potential therapeutic agent for OSCC.

Hierarchical Representation for Chromatic Processing across Macaque V1, V2, and V4.

The perception of color is an internal label for the inferred spectral reflectance of visible surfaces. To study how spectral representation is transformed through modular subsystems of successive cortical areas, we undertook simultaneous optical imaging of intrinsic signals in macaque V1, V2, and V4, supplemented by higher-resolution electrophysiology and two-photon imaging in awake macaques. We find a progressive evolution in the scale and precision of chromotopic maps, expressed by a uniform blob-like architecture of hue responses within each area. Two-photon imaging reveals enhanced hue-specific cell clustering in V2 compared with V1. A phenomenon of endspectral (red and blue) responses that is clear in V1, recedes in V2, and is virtually absent in V4. The increase in mid- and extra-spectral hue representations through V2 and V4 reflects the nature of hierarchical processing as higher areas read out locations in chromatic space from progressive integration of signals relayed by V1.

Immediate Impact of Acute Elevation of Intraocular Pressure on Cortical Visual Motion Processing.

Purpose: To physiologically examine the impairment of cortical sensitivity to visual motion during acute elevation of intraocular pressure (IOP). Methods: Motion processing in the cat brain is well characterized, its X and Y cell visual pathways being functionally analogous to parvocellular and magnocellular pathways in primates. Using this model, we performed ocular anterior chamber perfusion to reversibly elevate IOP over a range from 30 to 90 mm Hg while monitoring cortical activity with intrinsic signal optical imaging. Drifting random-dot fields and gratings were used to characterize cortical population responses to motion direction and orientation in early visual areas 17 and 18. Results: We found that acute IOP elevations at 50 mm Hg and above, which is often observed in acute glaucoma, suppressed cortical motion direction responses. This suppression was more profound in area 17 than in area 18, and more profound in central than peripheral visual field (eccentricities 0°-4° vs. 4°-8°) within area 17. In addition, orientation responses were more suppressed than motion direction responses for the same IOP modulation. Conclusions: In contrast to human chronic glaucoma that may cause greater dysfunction in large-cell magnocellular than in small-cell parvocellular visual pathways, our direct measurement of cortical processing networks implies that the small X-cell pathway shows greater vulnerability to acute IOP elevation than the large Y-cell pathway in visual motion processing. The results demonstrate that fine discrimination mechanisms for motion in the central visual field are particularly impacted by acute IOP attacks, suggesting a neural basis for immediate visual deficits in the fine motion perception of acute glaucoma patients.

The Serpentine Illusion: A Visual Motion Illusion Induced by Phase-Shifted Line Gratings.

In a pattern of horizontal lines containing ± 45° zigzagging phase-shifted strips, vivid illusory motion is perceived when the pattern is translated up or down at a moderate speed. Two forms of illusory motion are seen: [i] a motion "racing" along the diagonal interface between the strips and [ii] lateral (sideways) motion of the strip sections. We found the relative salience of these two illusory motions to be strongly influenced by the vertical spacing and length of the line gratings, and the period length of the zigzag strips. Both illusory motions are abolished when the abutting strips are interleaved, separated by a gap or when a real line is superimposed at the interface. Illusory motion is also severely weakened when equiluminant colored grating lines are used. Illusory motion perception is fully restored at < 20% luminance contrast. Using adaptation, we find that line-ends alone are insufficient for illusory motion perception, and that both physical carrier motion and line orientation are required. We finally test a classical spatiotemporal energy model of V1 cells that exhibit direction tuning changes that are consistent with the direction of illusory motion. Taking this data together, we constructed a new visual illusion and surmise its origin to interactions of spatial and temporal energy of the lines and line-ends preferentially driving the magnocellular pathway.

Impact of acute intraocular pressure elevation on the visual acuity of non-human primates.

BACKGROUND: Glaucoma is the leading cause of irreversible blindness worldwide and elevated intraocular pressure (IOP) is an established risk factor. Visual acuity, the capacity for fine analysis of spatial frequency (SF) information, is relatively preserved in central vision until the later stages of chronic glaucoma. However, for acute glaucoma that is associated with sharp IOP elevation, how visual acuity is affected by acute IOP elevation remains unclear. METHODS: Using intrinsic-signal optical imaging of large areas of visual cortices V1 and V2 in seven rhesus macaques, visual acuity was directly examined during acute IOP elevation at 70 mmHg, a pressure often observed in acute angle-closure glaucoma. Acute IOP elevation was achieved by reversible monocular anterior chamber perfusions, and visual acuity was quantified by cortical population responses to various SFs ranging from 0.5-6 cycles/°. FINDINGS: Acute IOP elevation particularly depressed the ability of the visual cortex to register fine details (at high SFs referring to visual acuity), an effect that was progressively more severe toward the central visual field. These results completely contrast with long-term impairments present in chronic glaucoma. INTERPRETATION: Our results show that impairment of fine visual discrimination within the central visual field is the principal consequence of sharp IOP elevation, implicating relatively greater dysfunction in parvocellular pathways. This study provides direct cortical neural evidence for the immediate visual acuity impairment in acute glaucoma patients. FUND: National Natural Science Foundation of China, Chinese Academy of Sciences, Shanghai Committee of Science and Technology, and Shanghai Municipal Health Commission.

Revealing Detail along the Visual Hierarchy: Neural Clustering Preserves Acuity from V1 to V4.

How primates perceive objects along with their detailed features remains a mystery. This ability to make fine visual discriminations depends upon a high-acuity analysis of spatial frequency (SF) along the visual hierarchy from V1 to inferotemporal cortex. By studying the transformation of SF across macaque parafoveal V1, V2, and V4, we discovered SF-selective functional domains in V4 encoding higher SFs up to 12 cycles/°. These intermittent higher-SF-selective domains, surrounded by domains encoding lower SFs, violate the inverse relationship between SF preference and retinal eccentricity. The neural activities of higher- and lower-SF domains correspond to local and global features, respectively, of the same stimuli. Neural response latencies in high-SF domains are around 10 ms later than in low-SF domains, consistent with the coarse-to-fine nature of perception. Thus, our finding of preserved resolution from V1 into V4, separated both spatially and temporally, may serve as a connecting link for detailed object representation.

Focal Gain Control of Thalamic Visual Receptive Fields by Layer 6 Corticothalamic Feedback.

The projections between the thalamus and primary visual cortex (V1) are a key reciprocal neural circuit, relaying retinal signals to cortical layers 4 & 6 while being simultaneously regulated by massive layer 6 corticothalamic feedback. Effectively dissecting the influence of this corticothalamic feedback circuit in higher mammals remains a challenge for vision research. By pharmacologically increasing the focal gain of visually driven layer 6 responses of cat V1 in a controlled fashion, we examined the effects of such focal cortical changes on the response amplitudes and spatial structure of the receptive fields (RFs) of individual dorsal lateral geniculate nucleus (dLGN) cells. We found that enhancing visually driven cortical feedback could facilitate or suppress the overall responses of dLGN cells, and such an effect was linked to the orientation preference of the cortical neuron. Related to these selective retinotopic gain changes, enhanced feedback induced the RFs of dLGN cells to expand, contract or shift their spatial focus. Our results provide further evidence for a functional mechanism through which the cortex can selectively gate visual information flow from the thalamus back to the visual cortex.

Phylogeny, Pathogenicity, and Transmission of H5N1 Avian Influenza Viruses in Chickens.

We analyzed five H5N1 avian influenza viruses (AIVs) isolated from different birds in 2012 in China. Based on whole-genome sequences, we divided the viruses into four genotypes. The DKE26, GSE43, and DKE53 viruses belonged to Genotypes 1-3, respectively. The CKE93 and CKE96 viruses were classified into Genotype 4. Genotypes 1-3 correspond to the viruses containing the HA gene of clade 2.3.2, and Genotype 4 is the virus that bears the HA gene of clade 7.2. To better understand the pathogenicity and transmission of the viruses, we infected chickens with 103 EID50/0.1 ml GSE43 (clade 2.3.2) or CKE93 (clade 7.2) virus. Our results revealed that 6 of 7 specific-pathogen-free (SPF) chickens inoculated with GSE43 virus were dead before 7-day post-infection, but all the SPF chickens inoculated with CKE93 virus survived the infection. Both the GSE43 and CKE93 viruses replicated systemically in chickens. The virus titers of GSE43 virus in tested organs were obviously higher than those of CKE93 virus. Our results revealed that the pathogenicity and replication of GSE43 in chickens was much higher than those of CKE93. The GSE43 virus could transmit between chickens, but the CKE93 could not transmit between chickens by naïve contact. Therefore, different clades of H5N1 AIVs possessed variable pathogenicities and transmission abilities among chickens. Our study contributes to knowledge of pathogenic variations of prevalent H5N1 viruses.

Asymmetries of Dark and Bright Negative Afterimages Are Paralleled by Subcortical ON and OFF Poststimulus Responses.

Humans are more sensitive to luminance decrements than increments, as evidenced by lower thresholds and shorter latencies for dark stimuli. This asymmetry is consistent with results of neurophysiological recordings in dorsal lateral geniculate nucleus (dLGN) and primary visual cortex (V1) of cat and monkey. Specifically, V1 population responses demonstrate that darks elicit higher levels of activation than brights, and the latency of OFF responses in dLGN and V1 is shorter than that of ON responses. The removal of a dark or bright disc often generates the perception of a negative afterimage, and here we ask whether there also exist asymmetries for negative afterimages elicited by dark and bright discs. If so, do the poststimulus responses of subcortical ON and OFF cells parallel such afterimage asymmetries? To test these hypotheses, we performed psychophysical experiments in humans and single-cell/S-potential recordings in cat dLGN. Psychophysically, we found that bright afterimages elicited by luminance decrements are stronger and last longer than dark afterimages elicited by luminance increments of equal sizes. Neurophysiologically, we found that ON cells responded to the removal of a dark disc with higher firing rates that were maintained for longer than OFF cells to the removal of a bright disc. The ON and OFF cell asymmetry was most pronounced at long stimulus durations in the dLGN. We conclude that subcortical response strength differences between ON and OFF channels parallel the asymmetries between bright and dark negative afterimages, further supporting a subcortical origin of bright and dark afterimage perception.SIGNIFICANCE STATEMENT Afterimages are physiological aftereffects following stimulation of the eye, the study of which helps us to understand how our visual brain generates visual perception in the absence of physical stimuli. We report, for the first time to our knowledge, asymmetries between bright and dark negative afterimages elicited by luminance decrements and increments, respectively. Bright afterimages are stronger and last longer than dark afterimages. Subcortical neuronal recordings of poststimulus responses of ON and OFF cells reveal similar asymmetries with respect to response strength and duration. Our results suggest that subcortical differences between ON and OFF channels help explain intensity and duration asymmetries between bright and dark afterimages, supporting the notion of a subcortical origin of bright and dark afterimages.

Electrically Evoked Responses in the Rabbit Cortex Induced by Current Steering With Penetrating Optic Nerve Electrodes.

Purpose: Current steering is a neural stimulation strategy that uses simultaneous stimulation of adjacent electrodes to produce additional intermediate stimulation sites and thus improves spatial resolution. We investigated the feasibility of current steering using electrophysiological and computational methods after implanting paired penetrating electrodes into the rabbit's optic nerve (ON). Methods: Penetrating electrodes at different interelectrode distances were implanted into the ON and electrically evoked cortical potentials (EEPs) in V1 recorded with a 6 × 8 array. The current thresholds, EEP amplitudes, and spatial distributions were analyzed during current steering. Computational simulation studies were performed based on finite element models to calculate the area and spatial distribution of recruited ON fibers using a current steering stimulation strategy. Results: Threshold reduction and EEP amplitude enhancement were found with simultaneous stimulation of closely spaced electrode pairs. Spatially shifted cortical responses were achieved using current steering, whereas the amplitudes and spatial spreads of the responses were similar to that elicited by a single electrode. Computational simulations suggested that the centroid of the ON recruitment area could be modulated by current steering while the total recruitment area did not show any appreciable variability at a fixed current intensity. Conclusions: Current steering is a useful strategy to enhance the spatial resolution of an ON prosthesis without increasing the number of physical electrodes. This study provides useful information for optimizing the design of stimulation strategies with a penetrating ON prosthesis.

Inverted optical intrinsic response accompanied by decreased cerebral blood flow are related to both neuronal inhibition and excitation.

Negative hemodynamic response has been widely reported in blood oxygenation level-dependent (BOLD) functional magnetic resonance imaging studies, however its origin is still controversial. Optical intrinsic signal (OIS) imaging can be used to study brain activity by simultaneously recording hemodynamic signals at different wavelengths with high spatial resolution. In this study, we found transcorneal electrical stimulation (TcES) could elicit both positive OIS response (POR) and negative OIS response (NOR) in cats' visual cortex. We then investigated the property of this negative response to TcES and its relationship with cerebral blood flow (CBF) and neuronal activity. Results from laser speckle contrast imaging showed decreased CBF in the NOR region while increased CBF in the POR region. Both planar and laminar electrophysiological recordings in the middle (500-700 µm) cortical layers demonstrated that decreased and increased neuronal activities were coexisted in the NOR region. Furthermore, decreased neuronal activity was also detected in the deep cortical layers in the NOR region. This work provides evidence that the negative OIS together with the decreased CBF should be explained by mechanisms of both neuronal inhibition and excitation within middle cortical layers. Our results would be important for interpreting neurophysiological mechanisms underlying the negative BOLD signals.

Breaking cover: neural responses to slow and fast camouflage-breaking motion.

Primates need to detect and recognize camouflaged animals in natural environments. Camouflage-breaking movements are often the only visual cue available to accomplish this. Specifically, sudden movements are often detected before full recognition of the camouflaged animal is made, suggesting that initial processing of motion precedes the recognition of motion-defined contours or shapes. What are the neuronal mechanisms underlying this initial processing of camouflaged motion in the primate visual brain? We investigated this question using intrinsic-signal optical imaging of macaque V1, V2 and V4, along with computer simulations of the neural population responses. We found that camouflaged motion at low speed was processed as a direction signal by both direction- and orientation-selective neurons, whereas at high-speed camouflaged motion was encoded as a motion-streak signal primarily by orientation-selective neurons. No population responses were found to be invariant to the camouflage contours. These results suggest that the initial processing of camouflaged motion at low and high speeds is encoded as direction and motion-streak signals in primate early visual cortices. These processes are consistent with a spatio-temporal filter mechanism that provides for fast processing of motion signals, prior to full recognition of camouflage-breaking animals.

Spatial characteristics of evoked potentials elicited by a MEMS microelectrode array for suprachoroidal-transretinal stimulation in a rabbit.

BACKGROUND: Suprachoroidal-transretinal stimulation (STS) can potentially restore vision. This study investigated the spatial characteristics of cortical electrical evoked potentials (EEPs) elicited by STS. METHODS: A 4 × 4 thin-film platinum microelectrode stimulating array (200 µm electrode diameter and 400 µm center-to-center distance) was fabricated by a micro-electro-mechanical systems (MEMS) techniques and implanted into the suprachoroidal space of albino rabbits. RESULTS: The current threshold to elicit reliable EEPs by a single electrode was 41.6 ± 12.6 µA, corresponding to a 66.2 ± 20.1 µC · cm(-2) charge density per phase, which was lower than the reported safety limits. Spatially differentiated cortical responses could be evoked by STS through different rows or columns of electrical stimulation; furthermore, shifts in the location of the maximum cortical activities were consistent with cortical visuotopic maps; increasing the number of simultaneously stimulating electrodes increased the response amplitudes of EEPs and expanded the spatial spread as well. In addition, long-term implantation and electrical stimulation of the MEMS electrode array in suprachoroidal space are necessary to evaluate systematically the safety and biocompatibility of this approach. CONCLUSIONS: This study indicates that the STS approach by a MEMS-based platinum electrode array is a feasible alternative for visual restoration, and relatively high spatial discrimination may be achieved.

Orientation-cue invariant population responses to contrast-modulated and phase-reversed contour stimuli in macaque V1 and V2.

Visual scenes can be readily decomposed into a variety of oriented components, the processing of which is vital for object segregation and recognition. In primate V1 and V2, most neurons have small spatio-temporal receptive fields responding selectively to oriented luminance contours (first order), while only a subgroup of neurons signal non-luminance defined contours (second order). So how is the orientation of second-order contours represented at the population level in macaque V1 and V2? Here we compared the population responses in macaque V1 and V2 to two types of second-order contour stimuli generated either by modulation of contrast or phase reversal with those to first-order contour stimuli. Using intrinsic signal optical imaging, we found that the orientation of second-order contour stimuli was represented invariantly in the orientation columns of both macaque V1 and V2. A physiologically constrained spatio-temporal energy model of V1 and V2 neuronal populations could reproduce all the recorded population responses. These findings suggest that, at the population level, the primate early visual system processes the orientation of second-order contours initially through a linear spatio-temporal filter mechanism. Our results of population responses to different second-order contour stimuli support the idea that the orientation maps in primate V1 and V2 can be described as a spatial-temporal energy map.

Asymmetrical color filling-in from the nasal to the temporal side of the blind spot.

The physiological blind spot, corresponding to the optic disk in the retina, is a relatively large (6 × 8°) area in the visual field that receives no retinal input. However, we rarely notice the existence of it in daily life. This is because the blind spot fills in with the brightness, color, texture, and motion of the surround. The study of filling-in enables us to better understand the creative nature of the visual system, which generates perceptual information where there is none. Is there any retinotopic rule in the color filling-in of the blind spot? To find out, we used mono-colored and bi-colored annuli hugging the boundary of the blind spot. We found that mono-colored annuli filled in the blind spot uniformly. By contrast, bi-colored annuli, where one half had a given color, while the other half had a different one, filled in the blind spot asymmetrically. Specifically, the color surrounding the nasal half typically filled in about 75% of the blind spot area, whereas the color surrounding the temporal half filled in only about 25%. This asymmetry was dependent on the relative size of the half rings, but not the two colors used, and was absent when the bi-colored annulus was rotated by 90°. Here, the two colors on the upper and lower sides of the blind spot filled in the enclosed area equally. These results suggest that the strength of filling-in decreases with distance from the fovea consistent with the decrease of the cortical magnification factor.

MJ-66 induces malignant glioma cells G2/M phase arrest and mitotic catastrophe through regulation of cyclin B1/Cdk1 complex.

Malignant gliomas are among the most devastating cancers as they are resistant to many kinds of treatment. Despite recent advances in the diagnosis and treatment, the prognosis of patients remains very poor and the development of new drug is urgently needed. Here, we report that a synthetic quinazolinone analog 2-(naphthalene-1-yl)-6-pyrrolidinyl-4-quinazolinone (MJ-66) induced glioma cell death. Immunofluorescence staining showed that MJ-66-induced cell death was associated with multinucleated phenotype and multipolar spindles that were typical characteristics of mitotic catastrophe. Flow cytometry analysis revealed that MJ-66 caused glioma cell cycle arrest at G2/M phase and increased the proportion of polyploidy cells. Western blotting indicated that the expression of cyclin B1, Cdk1 pY15 and Cdk1 increased after treatment with MJ-66. MJ-66 effectively inhibited tumor growth and induced apoptosis in the xenograft animal model of U87 human glioma cells. Together, these results suggest that MJ-66 inhibited malignant gliomas growth through inducing mitotic catastrophe by interference with G2/M cell cycle checkpoint which may open a new avenue for the treatment of malignant gliomas.

Optical imaging of visual cortical responses evoked by transcorneal electrical stimulation with different parameters.

PURPOSE: The use of phosphenes evoked by transcorneal electrical stimulation (TcES) has been proposed as a means of residual visual function evaluation and candidate selection before implantation of retinal prostheses. Compared to the subjective measures, measurement of neuronal activity in visual cortex can objectively and quantitatively explore their response properties to electrical stimulation. The purpose of this study was to investigate systematically the properties of cortical responses evoked by TcES. METHODS: The visual cortical responses were recorded using a multiwavelength optical imaging of intrinsic signals (OIS) combining with electrophysiological recording by a multichannel electrode array. The effects of different parameters of TcES on cortical responses, including the changes of hemoglobin oxygenation and cerebral blood volume, were examined. RESULTS: We found consistent OIS activation regions in visual cortex after TcES, which also showed strong evoked field potentials according to electrophysiological results. The OIS response regions were located mainly in cortical areas representing peripheral visual field. The extent of activation areas and strength of intrinsic signals were increased with higher current intensities and longer pulse widths, and the largest responses were acquired in the frequency range 10 to 20 Hz. CONCLUSIONS: Use of TcES through the ERG-jet corneal electrode may preferentially activate peripheral retina. Revealing the hemodynamic changes in visual cortex occurred after electrical stimulation can contribute to comprehension of neurophysiological underpinnings underlying prosthetic vision. This study provided an objective foundation for optimizing parameters of TcES and would bring considerable benefits in the application of TcES for assessment and screening in patients.

Electrical stimulation with a penetrating optic nerve electrode array elicits visuotopic cortical responses in cats.

OBJECTIVE: A visual prosthesis based on penetrating electrode stimulation within the optic nerve (ON) is a potential way to restore partial functional vision for blind patients. We investigated the retinotopic organization of ON stimulation and its spatial resolution. APPROACH: A five-electrode array was inserted perpendicularly into the ON or a single electrode was advanced to different depths within the ON (~1-2 mm behind the eyeball, 13 cats). A sparse noise method was used to map ON electrode position and the visual cortex. Cortical responses were recorded by a 5 × 6 array. The visuotopic correspondence between the retinotopic position of the ON electrode was compared with the visual evoked cortical map and the electrical evoked potentials elicited in response to ON stimulation. MAIN RESULTS: Electrical stimulation with penetrating ON electrodes elicited cortical responses in visuotopographically corresponding areas of the cortex. Stimulation of the temporal side of the ON elicited cortical responses corresponding to the central visual field. The visual field position shifted from the lower to central visual field as the electrode penetrated through the depth of the ON. A spatial resolution of ~ 2° to 3° within a limited cortical visuotopic representation could be obtained by this approach. SIGNIFICANCE: Visuotopic electrical stimulation with a relatively fine spatial resolution can be accomplished using penetrating electrodes implanted at multiple sites and at different depths within the ON just behind the globe. This study also provides useful experimental data for the design of electrode density and the distribution of penetrating ON electrodes for a visual prosthesis.