Connexin 30 locally controls actin cytoskeleton and mechanical remodeling in motile astrocytes.

During brain maturation, astrocytes establish complex morphologies unveiling intense structural plasticity. Connexin 30 (Cx30), a gap-junction channel-forming protein expressed postnatally, dynamically regulates during development astrocyte morphological properties by controlling ramification and extension of fine processes. However, the underlying mechanisms remain unexplored. Here, we found in vitro that Cx30 interacts with the actin cytoskeleton in astrocytes and inhibits its structural reorganization and dynamics during cell migration. This translates into an alteration of local physical surface properties, as assessed by correlative imaging using stimulated emission depletion (STED) super resolution imaging and atomic force microscopy (AFM). Specifically, Cx30 impaired astrocyte cell surface topology and cortical stiffness in motile astrocytes. As Cx30 alters actin organization, dynamics, and membrane physical properties, we assessed whether it controls astrocyte migration. We found that Cx30 reduced persistence and directionality of migrating astrocytes. Altogether, these data reveal Cx30 as a brake for astrocyte structural and mechanical plasticity.

Non-cell Autonomous OTX2 Homeoprotein Regulates Visual Cortex Plasticity Through Gadd45b/g.

The non-cell autonomous transfer of OTX2 homeoprotein transcription factor into juvenile mouse cerebral cortex regulates parvalbumin interneuron maturation and critical period timing. By analyzing gene expression in primary visual cortex of wild-type and Otx2+/GFP mice at plastic and nonplastic ages, we identified several putative genes implicated in Otx2-dependent visual cortex plasticity for ocular dominance. Cortical OTX2 infusion in juvenile mice induced Gadd45b/g expression through direct regulation of transcription. Intriguingly, a reverse effect was found in the adult, where reducing cortical OTX2 resulted in Gadd45b/g upregulation. Viral expression of Gadd45b in adult visual cortex directly induced ocular dominance plasticity with concomitant changes in MeCP2 foci within parvalbumin interneurons and in methylation states of several plasticity gene promoters, suggesting epigenetic regulation. This interaction provides a molecular mechanism for OTX2 to trigger critical period plasticity yet suppress adult plasticity.

A Mouse Model for Conditional Secretion of Specific Single-Chain Antibodies Provides Genetic Evidence for Regulation of Cortical Plasticity by a Non-cell Autonomous Homeoprotein Transcription Factor.

During postnatal life the cerebral cortex passes through critical periods of plasticity allowing its physiological adaptation to the environment. In the visual cortex, critical period onset and closure are influenced by the non-cell autonomous activity of the Otx2 homeoprotein transcription factor, which regulates the maturation of parvalbumin-expressing inhibitory interneurons (PV cells). In adult mice, the maintenance of a non-plastic adult state requires continuous Otx2 import by PV cells. An important source of extra-cortical Otx2 is the choroid plexus, which secretes Otx2 into the cerebrospinal fluid. Otx2 secretion and internalization requires two small peptidic domains that are part of the DNA-binding domain. Thus, mutating these "transfer" sequences also modifies cell autonomous transcription, precluding this approach to obtain a cell autonomous-only mouse. Here, we develop a mouse model with inducible secretion of an anti-Otx2 single-chain antibody to trap Otx2 in the extracellular milieu. Postnatal secretion of this single-chain antibody by PV cells delays PV maturation and reduces plasticity gene expression. Induced adult expression of this single-chain antibody in cerebrospinal fluid decreases Otx2 internalization by PV cells, strongly induces plasticity gene expression and reopens physiological plasticity. We provide the first mammalian genetic evidence for a signaling mechanism involving intercellular transfer of a homeoprotein transcription factor. Our single-chain antibody mouse model is a valid strategy for extracellular neutralization that could be applied to other homeoproteins and signaling molecules within and beyond the nervous system.

The hemodynamic signal as a first-order low-pass temporal filter: Evidence and implications for neuroimaging studies.

Neuronal activation triggers local changes in blood flow and hemoglobin oxygenation. These hemodynamic signals can be recorded through functional magnetic resonance imaging or intrinsic optical imaging, and allows inferring neural activity in response to stimuli. These techniques are widely used to uncover functional brain architectures. However, their accuracy suffers from distortions inherent to hemodynamic responses and noise. The analysis of these signals currently relies on models of impulse hemodynamic responses to brief stimuli. Here, in order to infer precise functional architectures, we focused on integrated signals associated to the dynamic response of functional maps. To this end, we recorded orientation and direction maps in cat primary visual cortex and compared two protocols: the conventional episodic stimulation technique and a continuous, periodic stimulation paradigm. Conventional methods show that the dynamics of activation and deactivation of the functional maps follows a linear first-order differential equation representing a low-pass filter. Comparison with the periodic stimulation methods confirmed this observation: the phase shifts and magnitude attenuations extracted at various frequencies were consistent with a low-pass filter with a 5s time constant. This dynamics presumably reflects the variations in deoxyhemoglobin mediated by arterial dilations. This dynamics open new avenues in the analysis of neuroimaging data that differs from common methods based on the hemodynamic response function. In particular, we demonstrate that inverting this first-order low-pass filter minimized the distortions of the signal and enabled a much faster and accurate reconstruction of functional maps.

Pinwheel-dipole configuration in cat early visual cortex.

In the early visual cortex, information is processed within functional maps whose layouts are thought to underlie visual perception. However, the precise organization of these functional maps as well as their interrelationships remain unsettled. Here, we show that spatial frequency representation in cat early visual cortex exhibits singularities around which the map organizes like an electric dipole potential. These singularities are precisely co-located with singularities of the orientation map: the pinwheel centers. To show this, we used high resolution intrinsic optical imaging in cat areas 17 and 18. First, we show that a majority of pinwheel centers exhibit in their neighborhood both semi-global maximum and minimum in the spatial frequency map (i.e. extreme values of the spatial frequency in a hypercolumn). This contradicts pioneering studies suggesting that pinwheel centers are placed at the locus of a single spatial frequency extremum. Based on an analogy with electromagnetism, we proposed a mathematical model for a dipolar structure, accurately fitting optical imaging data. We conclude that a majority of orientation pinwheel centers form spatial frequency dipoles in cat early visual cortex. Given the functional specificities of neurons at singularities in the visual cortex, it is argued that the dipolar organization of spatial frequency around pinwheel centers could be fundamental for visual processing.

Parsimony, Exhaustivity and Balanced Detection in Neocortex.

The layout of sensory brain areas is thought to subtend perception. The principles shaping these architectures and their role in information processing are still poorly understood. We investigate mathematically and computationally the representation of orientation and spatial frequency in cat primary visual cortex. We prove that two natural principles, local exhaustivity and parsimony of representation, would constrain the orientation and spatial frequency maps to display a very specific pinwheel-dipole singularity. This is particularly interesting since recent experimental evidences show a dipolar structures of the spatial frequency map co-localized with pinwheels in cat. These structures have important properties on information processing capabilities. In particular, we show using a computational model of visual information processing that this architecture allows a trade-off in the local detection of orientation and spatial frequency, but this property occurs for spatial frequency selectivity sharper than reported in the literature. We validated this sharpening on high-resolution optical imaging experimental data. These results shed new light on the principles at play in the emergence of functional architecture of cortical maps, as well as their potential role in processing information.

Organization and origin of spatial frequency maps in cat visual cortex.

It remains controversial whether and how spatial frequency (SF) is represented tangentially in cat visual cortex. Several models were proposed, but there is no consensus. Worse still, some data indicate that the SF organization previously revealed by optical imaging techniques simply reflects non-stimulus-specific responses. Instead, stimulus-specific responses arise from the homogeneous distribution of geniculo-cortical afferents representing X and Y pathways. To clarify this, we developed a new imaging method allowing rapid stimulation with a wide range of SFs covering more than 6 octaves with only 0.2 octave resolution. A benefit of this method is to avoid error of high-pass filtering methods which systematically under-represent dominant selectivity features near pinwheel centers. We show unequivocally that SF is organized into maps in cat area 17 (A17) and area 18 (A18). The SF organization in each area displays a global anteroposterior SF gradient and local patches. Its layout is constrained to that of the orientation map, and it is suggested that both maps share a common functional architecture. A17 and A18 are bound at the transition zone by another SF gradient involving the geniculo-cortical and the callosal pathways. A model based on principal component analysis shows that SF maps integrate three different SF-dependent channels. Two of these reflect the segregated excitatory input from X and Y geniculate cells to A17 and A18. The third one conveys a specific combination of excitatory and suppressive inputs to the visual cortex. In a manner coherent with anatomical and electrophysiological data, it is interpreted as originating from a subtype of Y geniculate cells.

Astroglial networks control visual responses of superior collicular neurons and sensory-motor behavior.

Astroglial networks closely interact with neuronal populations, but their functional contribution to neuronal representation of sensory information remains unexplored. The superior colliculus (SC) integrates multi-sensory information by generating distinct spatial patterns of neuronal functional responses to specific sensory stimulation. Here, we report that astrocytes from the mouse SC form extensive networks in the retinorecipient layer compared to visual cortex. This strong astroglial connectivity relies on high expression of gap-junction proteins. Genetic disruption of this connectivity functionally impairs SC retinotopic and orientation preference responses. These alterations are region specific, absent in primary visual cortex, and associated at the circuit level with a specific impairment of collicular neurons synaptic transmission. This has implications for SC-related visually induced innate behavior, as disrupting astroglial networks impairs light-evoked temporary arrest. Our results indicate that astroglial networks shape synaptic circuit activity underlying SC functional visual responses and play a crucial role in integrating visual cues to drive sensory-motor behavior.

Parallel development of orientation maps and spatial frequency selectivity in cat visual cortex.

In an early stage of the postnatal development of cats, orientation maps mature and spatial frequency selectivity is consolidated. To investigate the time course of orientation map maturation associated with the consolidation of spatial frequency selectivity, we performed optical imaging of intrinsic signals in areas 17 and 18 of cats under the stimulation of drifting square-wave gratings with different orientations and spatial frequencies. First, orientation maps for lower spatial frequencies emerged in the entire part of the lateral gyrus, which includes areas 17 and 18, and then these orientation maps in the posterior part of the lateral gyrus disappeared as orientation maps for higher spatial frequencies matured. Independent of age, an anteroposterior gradient of response strengths from lower to higher spatial frequencies was observed. This indicates that the regional distribution of spatial frequencies is innately determined. The size of iso-orientation domains tended to decrease as the stimulus spatial frequency increased at every age examined. In contrast, orientation representation bias changed with age. In cats younger than 3 months, the cardinal (vertical and horizontal) orientations were represented predominantly over the oblique orientations. However, in young adult cats from 3 to 9 months old, the representation bias switched to predominantly oblique orientations. These age-dependent changes in the orientation representation bias imply that orientation maps continue to elaborate within postnatal 1 year with the consolidation of spatial frequency selectivity. We conclude that both intrinsic and mutual factors lead to the development of orientation maps and spatial frequency selectivity.

Astrocytes close the mouse critical period for visual plasticity.

Brain postnatal development is characterized by critical periods of experience-dependent remodeling of neuronal circuits. Failure to end these periods results in neurodevelopmental disorders. The cellular processes defining critical-period timing remain unclear. Here, we show that in the mouse visual cortex, astrocytes control critical-period closure. We uncover the underlying pathway, which involves astrocytic regulation of the extracellular matrix, allowing interneuron maturation. Unconventional astrocyte connexin signaling hinders expression of extracellular matrix-degrading enzyme matrix metalloproteinase 9 (MMP9) through RhoA-guanosine triphosphatase activation. Thus, astrocytes not only influence the activity of single synapses but also are key elements in the experience-dependent wiring of brain circuits.

Development and Reorganization of Orientation Representation in the Cat Visual Cortex: Experience-Dependent Synaptic Rewiring in Early Life.

To date, numerous mathematical models have been proposed on the basis of some types of Hebbian synaptic learning to account for the activity-dependent development of orientation maps as well as neuronal orientation selectivity. These models successfully reproduced orientation map-like spatial patterns. Nevertheless, we still have questions: (1) How does synaptic rewiring occur in the visual cortex during the formation of orderly orientation maps in early life? (2) How does visual experience contribute to the maturation of orientation selectivity of visual cortical neurons and reorganize orientation maps? (3) How does the sensitive period for orientation plasticity end? In this study, we performed animal experiments and mathematical modeling to understand the mechanisms underlying synaptic rewiring for experience-dependent formation and reorganization of orientation maps. At first, we visualized orientation maps from the intrinsic signal optical imaging in area 17 of kittens reared under single-orientation exposure through cylindrical-lens-fitted goggles. The experiments revealed that the degree of expansion of cortical domains representing the experienced orientation depends on the age at which the single-orientation exposure starts. As a result, we obtained the sensitive period profile for orientation plasticity. Next, we refined our previously proposed mathematical model for the activity-dependent self-organization of thalamo-cortical inputs on the assumption that rewiring is caused by the competitive interactions among transient synaptic contacts on the same dendritic spine. Although various kinds of molecules have been reported to be involved in such interactions, we attempt to build a mathematical model to describe synaptic rewiring focusing on brain-derived neurotrophic factor (BDNF) and its related molecules. Performing computer simulations based on the refined model, we successfully reproduced orientation maps reorganized in kittens reared under single-orientation exposure as well as normal visual experience. We also reproduced the experimentally obtained sensitive period profile for orientation plasticity. The excellent agreement between experimental observations and theoretical reproductions suggests that the BDNF-induced competitive interaction among synaptic contacts from different axons on the same spine is an important factor for the experience-dependent formation and reorganization of orientation selectivity and orientation maps.

Anisotropy in the representation of direction preferences in cat area 18.

Higher visual cortical areas are involved in the perception of complex stimuli, such as the optic flow created by self-motion. On the other hand, area 18 is thought to extract primitive visual features, feeding higher cortical areas for further processing. In this study, we applied optical imaging of intrinsic signals in the central, lower visual field of cat area 18, and reconstructed direction preference and direction selectivity maps in each hemisphere. We observed a significant overrepresentation of downward and temporal directions, in accordance with previous electrophysiological results. Cardinal orientations were not overrepresented, however. Downward directions were overrepresented at the highest direction selectivity domains. Temporal direction representation, on the other hand, decreased with direction selectivity. Our findings therefore suggest the existence of a neural substrate for the processing of optic flow in cat area 18.

Chronically mountable goggles for persistent exposure to single orientation.

To examine the effect of experience on the developmental plasticity of functional maps in the visual cortex, we need to establish a method for a stable visual experience manipulation under the freely moving condition. For this purpose, we fabricated goggles that are chronically mounted stably on the animal's head, but easy to replace according to the animal's growth. Here we report the design of the goggles and the method of mounting them on the head of animals. By this method, combined with the intrinsic signal optical imaging technique, we were able to observe a rapid and robust reorganization of orientation maps.

Online analysis method for intrinsic signal optical imaging.

The intrinsic optical imaging technique has been widely applied for the visualization of functional maps in the sensory cortices of mammals. Many current studies refer this mapping in order to focus thereafter on particular features, at some particular locations: a fast and accurate mapping is therefore required. However, even during a successful experiment, the recorded raw data are usually contaminated by some kinds of noise that cannot necessarily be averaged out over the trials. An adequate image data analysis method has to be applied to extract signals closely related neural activities in response to presented stimuli. Thus far two different analysis methods could be adopted: the band-pass filtering and the GIF method [Yokoo T, Knight BW, Sirovich L. An optimization approach to signal extraction from noisy multivariate data. NeuroImage 2001:14;1309-26]. While the latter one is very efficient but requires the whole data in order to maximize the signal to noise ratio, the simple band-pass filtering technically reaches its limits very quickly. Here we propose another filtering method based on the polynomial subtraction of spatially smoothly modulated components. This simple method can visualize well-organized iso-orientation domains of the cat visual cortex with reliability similar to more sophisticated ones while allowing an online visualization of the clean data.

Roles of visual experience and intrinsic mechanism in the activity-dependent self-organization of orientation maps: theory and experiment.

It is widely accepted that functional maps in the mammalian visual cortex such as ocular dominance columns and orientation columns are formed depending on neural activity. There is still, however, controversy on how much visual experience contributes to the map formation during development. In the present study, we address this issue from mathematical modeling and experimental investigation. Using a model of activity-dependent self-organization of geniculo-cortical afferent inputs, we showed that spontaneous activity in the LGN can produce orientation maps, while the exposure to drifting gratings results in sharply segregated orientation maps as observed in cat visual cortex. The restricted exposure to a single orientation of the grating led to the over-representation of the exposed orientation, which was moderated by the contribution of learning based on the spontaneous activity. These theoretical results were confirmed by intrinsic optical recordings from area 18 of kittens reared under various visual conditions.

Postural control in children with strabismus: effect of eye surgery.

The purpose of this study was to examine the postural control in children with strabismus before and after eye surgery. Control of posture is a complex multi-sensorial process relying on visual, vestibular and proprioceptive systems. Reduced influence of one of such systems leads to postural adaptation due to a compensation of one of the other systems [3]. Nine children with strabismus (4-8 years old) participated in the study. Ophthalmologic, orthoptic, vestibular and postural tests were done before and twice (2 and 8 weeks) after eye surgery. Postural stability was measured by a platform (TechnoConcept): two components of the optic flux were used for stimulation (contraction and expansion) and two conditions were tested eyes open and eyes closed. The surface area of the center of pressure (CoP), the variance of speed of the CoP and the frequency spectrum of the platform oscillations by fast Fourier transformation were analysed. Before surgery, similar to typically developing children, postural stability was better in the eyes open condition. The frequency analysis revealed that for the low frequency band more energy was spent in the antero-posterior direction compared to the medio-lateral one while the opposite occurred for the middle and the high frequency bands. After surgery, the eye deviation was reduced in all children and their postural stability also improved. However, the energy of the high frequency band in the medio-lateral direction increased significantly. These findings suggest that eye surgery influences somatosensory properties of extra-ocular muscles leading to improvement of postural control and that binocular visual perception could influence the whole body.

Asymmetrical interhemispheric connections develop in cat visual cortex after early unilateral convergent strabismus: anatomy, physiology, and mechanisms.

In the mammalian primary visual cortex, the corpus callosum contributes to the unification of the visual hemifields that project to the two hemispheres. Its development depends on visual experience. When this is abnormal, callosal connections must undergo dramatic anatomical and physiological changes. However, data concerning these changes are sparse and incomplete. Thus, little is known about the impact of abnormal postnatal visual experience on the development of callosal connections and their role in unifying representation of the two hemifields. Here, the effects of early unilateral convergent strabismus (a model of abnormal visual experience) were fully characterized with respect to the development of the callosal connections in cat visual cortex, an experimental model for humans. Electrophysiological responses and 3D reconstruction of single callosal axons show that abnormally asymmetrical callosal connections develop after unilateral convergent strabismus, resulting from an extension of axonal branches of specific orders in the hemisphere ipsilateral to the deviated eye and a decreased number of nodes and terminals in the other (ipsilateral to the non-deviated eye). Furthermore this asymmetrical organization prevents the establishment of a unifying representation of the two visual hemifields. As a general rule, we suggest that crossed and uncrossed retino-geniculo-cortical pathways contribute successively to the development of the callosal maps in visual cortex.

A postnatal critical period for orientation plasticity in the cat visual cortex.

Orientation selectivity of primary visual cortical neurons is an important requisite for shape perception. Although numerous studies have been previously devoted to a question of how orientation selectivity is established and elaborated in early life, how the susceptibility of orientation plasticity to visual experience changes in time remains unclear. In the present study, we showed a postnatal sensitive period profile for the modifiability of orientation selectivity in the visual cortex of kittens reared with head-mounted goggles for stable single-orientation exposure. When goggle rearing (GR) started at P16-P30, 2 weeks of GR induced a marked over-representation of the exposed orientation, and 2 more weeks of GR consolidated the altered orientation maps. GR that started later than P50, in turn, induced the under-representation of the exposed orientation. Orientation plasticity in the most sensitive period was markedly suppressed by cortical infusion of NMDAR antagonist. The present study reveals that the plasticity and consolidation of orientation selectivity in an early life are dynamically regulated in an experience-dependent manner.

Orientation-restricted continuous visual exposure induces marked reorganization of orientation maps in early life.

To elucidate the effect of visual experience on the development of orientation maps, we conducted intrinsic signal optical imaging of the visual cortex of kittens that were continuously exposed to a single orientation through cylindrical-lens-fitted goggles under a freely moving condition starting at post-natal week 3. We observed a rapid reorganization of orientation maps, characterized by extensive representation of exposed orientations with reduced responsiveness to unexposed orientations. The over-representation of exposed orientation was marked for 1-2 weeks of goggle rearing. A longer period of goggle rearing, however, decreased the degree of over-representation, which still remained at a remarkable level. Dark rearing episodes daily interleaved between single orientation exposures moderated the over-representation effect. Unit recording from goggle-reared kittens showed preferred orientations consistent with optical imaging. Using c-Fos immunoreactivity mapping, we showed that the number of neurons strongly responding to the exposed orientation was 3 times larger in a goggle-reared cat than the number of neurons responding to the vertical orientation in a normal cat. Taken together, these results suggest that the reorganization of orientation maps was caused by the expansion of domains maximally responding to exposed orientation as well as the strong reduction of responses to unexposed orientations.