A distrofina é uma das proteínas que formam o complexo glicoproteico necessário para a integridade da fibra muscular e sua disfunção causa uma doença genética letal para os seres humanos, a distrofia muscular de Duchenne (DMD). Além do papel fundamental no tecido muscular, a distrofina é necessária para a fisiologia da retina e, portanto, para o processamento da informação visual. Estudos anteriores mostraram prejuízo assimétrico no eletrorretinograma (ERG), maior para aumento da luminância (via ON) que para diminuição (via OFF). Além disso, prejuízos na visão de cores e contrastes eram mais frequentes e severos em pacientes com alterações genéticas que comprometem a expressão da isoforma Dp260. O objetivo do presente estudo foi verificar através de protocolos eletrofisiológicos e psicofísicos se existiam diferenças nas respostas mediadas pelas vias visuais ON e OFF em jovens com DMD e como estas se relacionavam com o genótipo. Foram avaliados 19 jovens com DMD (idade média = 15,2 ± 3,4 anos) cujos resultados foram comparados com os de sujeitos controles pareados por idade. Os métodos utilizados foram o ERG de campo total e medidas psicofísicas de sensibilidade ao contraste (SC) espacial e temporal de luminância. Protocolos tradicionalmente empregados foram associados a protocolos cujos estímulos visuais ativam, preferencialmente, a via ON ou a via OFF. Para o ERG de campo total foram utilizados seis protocolos: 1. ERG escotópico, 2. ERG fotópico, 3 e 4. ERG mesópico ON e OFF, 5 e 6. ERG fotópico ON e OFF. Para os quatro últimos foram utilizados estímulos intermitentes com modulação da luminância em dente de serra, com aumento rápido de luminância e diminuição gradual (ON) e o contrário (OFF). Para a avaliação psicofísica foi determinada: 1. SC para grades senoidais e SC temporal, e 2. SC a estímulos de tabuleiro de xadrez com aumento (ON) ou diminuição (OFF) da luminância média relativa ao fundo, apresentados com duração curta (sistema magnocelular) ou longa (sistema parvocelular). Os resultados mostraram redução da amplitude da onda-b dos ERGs escotópico e fotópico e prejuízos na SC espacial e temporal de luminância, concordando com a literatura. A contribuição inédita do presente estudo foi mostrar alteração nos ERGs ON e OFF para atividade dos bastonetes e no ERG ON para atividade exclusiva dos cones. Na avaliação psicofísica, houve redução da SC para os protocolos ON sem diferença entre magnocelular e parvocelular. Em conclusão, as alterações encontradas estão principalmente relacionadas com a atividade ON da retina. A alteração psicofísica da SC espacial de luminância de jovens com DMD deve estar relacionada, ao menos em parte, com prejuízos retinianos devidos à ausência da Dp260 ou da própria distrofina total (Dp427). Estudos futuros devem aprofundar a investigação utilizando protocolos do ERG que estimulam, preferencialmente, as vias magnocelular e parvocelular, e ampliar o número de pacientes avaliados para se obter as correlações entre as alterações genéticas e os prejuízos visuais / The dystrophin is one of the proteins that form the glycoprotein complex necessary for the integrity of the muscular fibers. Its dysfunction causes a genetic disease called the Duchenne Muscular Dystrophy (DMD) which is lethal for humans. Besides its fundamental role in the muscle tissues the dystrophin is also necessary in the physiology of the retina, in the processing of visual information. Previous studies have demonstrated asymmetric deviations in the electroretinogram (ERG) of DMDs with bigger changes to stimuli with increasing (ON pathway) than to decreasing (OFF pathway) luminance. Moreover deficiencies in color vision and contrast sensitivity have been more frequent and severe in patients having genetic alterations related to the expression of the Dp260 isoform. The aim of the present study was to apply electrophysiological and psychophysical protocols to verify the suspected alterations in DMD patients regarding the ON and OFF visual pathways and relate the results to their genotypes. 19 DMD patients (mean age = 15.2 ± 3.4 years) were tested and their results were compared to that of an age-matched control group. Fullfield ERG and spatial and temporal luminance contrast sensitivity tests were used during the examinations. Classical protocols were applied together with the ones preferentially stimulating the ON and the OFF visual pathways. The full-field ERG test consisted of six protocols: 1. Scotopic ERG, 2. Photopic ERG, 3. and 4. Mesopic ON and OFF ERG, 5. and 6. Photopic ON and OFF ERG. For the latter four protocols flicker stimuli were used with sawtooth modulation of rapid increase and slower decrease in luminance (ON) and rapid decrease and slower increase in luminance (OFF). The psychophysical evaluation comprised 1. Spatial contrast sensitivity test with sinusoidal gratings and temporal contrast sensitivity test, and 2. Contrast sensitivity tests with checkerboard stimuli with increasing (ON) and decreasing (OFF) luminance relative to the background. These latter were presented for both short (magnocellular system) and long (parvocellular system) durations. In agreement with the literature the results show reduced amplitudes in the ERGs scotopic and photopic b-waves and also impairment in the spatial and temporal contrast sensitivities. This studys novel contribution was the presentation of the alterations in both rod driven ON and OFF ERGs and solely in the cone driven ON ERG. The psychophysical analysis showed reduced contrast sensitivity in the ON protocol similar for both magno- and parvocellular oriented stimuli. In summary the encountered alterations suggest damages in the ON mechanism of the retina. The changes in spatial luminance contrast sensitivity of DMD patients are related, at least partially, to the lack of Dp260 or to the loss of the entire dystrophin (Dp427). Future studies shall investigate this in more details applying ERG protocols to stimulate magno- and parvocellular activities, and increase the number of patients to be able to determine correlations between visual dysfunctions and genetic mutations

Functional MRI or Functional Magnetic Resonance Imaging (fMRI) is a type of specialized MRI scan which measures the hemodynamic response related to neural activity in the brain or spinal cord of humans and animals. Due to its relatively low invasiveness, absence of radiation exposure, and relatively wide availability, functional MRI has come to dominate the brain mapping field since the early 1990s. The objective of this thesis work is to develop and apply functional MRI methods at 7 Tesla, for in vivo investigation of rodent visual and auditory system. Firstly, the development of the rat visual pathway was studied by blood oxygenation level–dependent (BOLD) contrast from the time of eyelid opening (P14) to adulthood (P60) in normal rat brain. By studying BOLD-fMRI measurements in the normal brain superior colliculus (SC), we determined that the regional BOLD response undergoes a systematic increase in amplitude especially over the third postnatal week. Secondly, the potential for plasticity of the rodent superior colliculus (SC) was studied using BOLD fMRI. By studying BOLD-fMRI measurements in the SC of three groups of rats (normal, HI-injured with left SC partially damaged and HI-injured with left SC completely damaged), we can evaluate the extent of plastic changes, compensatory and transneuronal plasticity after varying degrees of SC injury. We also applied BOLD-fMRI using very short repetition time (TR) of 0.2s on rats to measure the difference in response temporal dynamics between the SC and LGN, which has not been measured conclusively or with high temporal resolution. The primary finding in this study is that there is an approximately 0.8s difference between the BOLD responses of the rat contralateral SC and LGN to the visual stimuli. In addition, the amplitude of the SC response is larger than that of the LGN. Thirdly, BOLD-fMRI is used to measure the SC hemodynamic responses, in normal adult Sprague-Dawley (SD) rats, during a dynamic visual stimulus similar to those used in long-range apparent motion studies. The stimulation paradigm mimic effective speeds of motion between 7 and 164?/s, the results suggest that the SC is sensitive to slow moving visual stimuli but the hemodynamic response is reduced at higher speeds. Finally, BOLD-fMRI is used to study hemodynamic response temporal dynamics in the superior colliculus (SC) and inferior colliculus (IC) following visual and auditory associated stimulation. Our results show the baselines of SC BOLD signal (in two sides) increase during the ON period of auditory stimulation, which demonstrate that auditory stimulation can increase ROI activation signal intensity in superior colliculus (SC). The previous dominant theory is that individual senses each have separate areas of the brain dedicated to processing each sense, while the individual sense perceptions are integrated together to produce a multi-sensory experience. As a result of new research over the past several years, however, this view has been challenged by studies showing that processing in the visual area of the brain can be directly influenced by hearing and touch. All these discoveries represent a new view of how the brain is actually organized. / published_or_final_version / Electrical and Electronic Engineering / Master / Master of Philosophy

Retinal ganglion cell (RGC) axon arbors within the optic tectum are refined in development through a dynamic process of activity-dependent remodeling. The synaptic adhesion molecule N-cadherin is a candidate for mediating selective stabilization and elaboration of RGC axons due to its localization to perisynaptic sites and its modifiability by neural activity. RGCs of Xenopus tadpoles were co-transfected with plasmids encoding a dominant negative N-cadherin (N-cadDeltaE) and eGFP or eYFP. Using two-photon in vivo time-lapse imaging, we found that axons expressing N-cadDeltaE became less elaborate than controls over three days of daily live imaging. Shorter interval time-lapse imaging of axons expressing synaptophysin-GFP to visualize putative synaptic sites revealed that N-cadDeltaE expressing axons form fewer stable branches than controls and that stabilization of axonal branches at synaptic sites is altered. We conclude that N-cadherin participates in the stabilization of axonal branches in the Xenopus retinotectal system.

[Truncated abstract] Developmentally, the rodent retinocollicular projection is often thought of as a homogenous projection of retinal ganglion cell (RGC) axons, however the extensive period of RGC neurogenesis and sequential arrival of their axons into central targets such as the superior colliulus (SC) suggests otherwise. RGC axons are already present in the developing SC at embryonic (E) day 16.5-17. RGCs born on E15 have innervated the SC by birth, whereas axons derived from RGCs that are born last (E19) do not grow into the SC until postnatal (P) days 4-6 (Dallimore et al., 2002). These observations may go someway to explaining why, after SC lesions in rats at P2, there is greater growth distal to the lesion site compared to lesions made at P6 (Tan and Harvey, 1997b). It may be that the post lesion growth is simply de novo growth of axons from late-born RGCs rather than regeneration of pre-existing, injured axons. Early and late cohorts of growing RGC axons presumably encounter different developmental terrains as they grow from retina to central targets, possibly resulting in differences in developmental milestones and growth potentials. There may also be differences in guidance cues, further suggesting that gene expression in early vs late born RGCs may differ. To examine differences between early (E15) and late (E19) born RGCs during development, the time-course and extent of programmed RGC death in normal rat pups, and RGC death following the removal of target-derived trophic factors, was assessed. ... On the other hand, LCM captured GCL analysed for gene expression at P0 and P7 revealed decreases in AKT, Math5, Notch1, c-jun, DCC, Arginase-1 mRNA levels and a considerable decrease in GAP-43 expression. It is not surprising to see differences in gene expression between whole eye and the more specific GCL samples, as the cells in all layers of the retina have very different functions and different developmental profiles. It is important to note decreases in mRNA expression in the GCL for a number of the genes analysed at P0 and P7, reflecting cessation of RGC death and completion of axonal growth into central visual targets. I also examined at the protein level expression of DCC, Arginase1, c-Jun and Bcl-2 at birth (P0) in BrdU labeled RGCs born on E15 or E19. When comparing the percentage of double labelled cells compared to the total number of cells expressing each protein, Bcl-2, c-Jun and Arg1 were expressed more in E15 RGCs (22.90%, 72.71%, and 16.44% respectively in E15 RGCs, compared with 0.52%, 13.17% and 3.59% in E19 RGCs). In contrast, DCC was expressed more at birth in E19 RGCs (18.05% in E19 RGCs compared with 9.23% in E15 RGCs). This shows there is clearly a difference in the expression of proteins in the two cohorts of RGCs, which is consistent with PCR data and with their growth state as their axons encounter the changes in the newborn brain. The overall findings of this research suggest that seemingly homogenous populations of neurons are quite different in their developmental profile and in their response to injury. This work may provide new ways of determining better strategies for CNS repair and the most effective way of targeting cells for regeneration and survival.

Until recently induced gamma-band activity (GBA) was considered a neural marker of cortical object representation. However, induced GBA in the electroencephalogram (EEG) is susceptible to artifacts caused by miniature fixational saccades. Recent studies have demonstrated that fixational saccades also reflect high-level representational processes. Do high-level as opposed to low-level factors influence fixational saccades? What is the effect of these factors on artifact-free GBA? To investigate this, we conducted separate eye tracking and EEG experiments using identical designs. Participants classified line drawings as objects or non-objects. To introduce low-level differences, contours were defined along different directions in cardinal color space: S-cone-isolating, intermediate isoluminant, or a full-color stimulus, the latter containing an additional achromatic component. Prior to the classification task, object discrimination thresholds were measured and stimuli were scaled to matching suprathreshold levels for each participant. In both experiments, behavioral performance was best for full-color stimuli and worst for S-cone isolating stimuli. Saccade rates 200–700 ms after stimulus onset were modulated independently by low and high-level factors, being higher for full-color stimuli than for S-cone isolating stimuli and higher for objects. Low-amplitude evoked GBA and total GBA were observed in very few conditions, showing that paradigms with isoluminant stimuli may not be ideal for eliciting such responses. We conclude that cortical loops involved in the processing of objects are preferentially excited by stimuli that contain achromatic information. Their activation can lead to relatively early exploratory eye movements even for foveally-presented stimuli.

No description available.

No / We identified human visual field maps, LO1 and LO2, in object-selective lateral occipital cortex. Using transcranial magnetic stimulation (TMS), we assessed the functions of these maps in the perception of orientation and shape. TMS of LO1 disrupted orientation, but not shape, discrimination, whereas TMS of LO2 disrupted shape, but not orientation, discrimination. This double dissociation suggests that specialized and independent processing of different visual attributes occurs in LO1 and LO2.

No description available.

In this report, we evaluate the role of visual areas responsive to motion in the human brain in the perception of stimulus speed. We first identified and localized V1, V3A, and V5/MT+ in individual participants on the basis of blood oxygenation level-dependent responses obtained in retinotopic mapping experiments and responses to moving gratings. Repetitive transcranial magnetic stimulation (rTMS) was then used to disrupt the normal functioning of the previously localized visual areas in each participant. During the rTMS application, participants were required to perform delayed discrimination of the speed of drifting or spatial frequency of static gratings. The application of rTMS to areas V5/MT and V3A induced a subjective slowing of visual stimuli and (often) caused increases in speed discrimination thresholds. Deficits in spatial frequency discrimination were not observed for applications of rTMS to V3A or V5/MT+. The induced deficits in speed perception were also specific to the cortical site of TMS delivery. The application of TMS to regions of the cortex adjacent to V5/MT and V3A, as well as to area V1, produced no deficits in speed perception. These results suggest that, in addition to area V5/MT+, V3A plays an important role in a cortical network that underpins the perception of stimulus speed in the human brain.