Experience dependent plasticity of higher visual cortical areas in the mouse.

Experience dependent plasticity in the visual cortex is a key paradigm for the study of mechanisms underpinning learning and memory. Despite this, studies involving manipulating visual experience have largely been limited to the primary visual cortex, V1, across various species. Here we investigated the effects of monocular deprivation (MD) on the ocular dominance (OD) and orientation selectivity of neurons in four visual cortical areas in the mouse: the binocular zone of V1 (V1b), the putative "ventral stream" area LM and the putative "dorsal stream" areas AL and PM. We employed two-photon calcium imaging to record neuronal responses in young adult mice before MD, immediately after MD, and following binocular recovery. OD shifts following MD were greatest in LM and smallest in AL and PM; in LM and AL, these shifts were mediated primarily through a reduction of deprived-eye responses, in V1b and LM through an increase in response through the non-deprived eye. The OD index recovered to pre-MD levels within 2 weeks in V1 only. MD caused a reduction in orientation selectivity of deprived-eye responses in V1b and LM only. Our results suggest that changes in OD in higher visual areas are not uniformly inherited from V1.

The separate and combined properties of the granular (area 29) and dysgranular (area 30) retrosplenial cortex.

Retrosplenial cortex contains two principal subdivisions, area 29 (granular) and area 30 (dysgranular). Their respective anatomical connections in the rat brain reveal that area 29 is the primary recipient of hippocampal and parahippocampal spatial and contextual information while area 30 is the primary interactor with current visual information. Lesion studies and measures of neuronal activity in rodents indicate that retrosplenial cortex helps to integrate space from different perspectives, e.g., egocentric and allocentric, providing landmark and heading cues for navigation and spatial learning. It provides a repository of scene information that, over time, becomes increasingly independent of the hippocampus. These processes, reflect the interactive actions between areas 29 and 30, along with their convergent influences on cortical and thalamic targets. Consequently, despite their differences, both areas 29 and 30 are necessary for an array of spatial and learning problems.

Stable Encoding of Visual Cues in the Mouse Retrosplenial Cortex.

The rodent retrosplenial cortex (RSC) functions as an integrative hub for sensory and motor signals, serving roles in both navigation and memory. While RSC is reciprocally connected with the sensory cortex, the form in which sensory information is represented in the RSC and how it interacts with motor feedback is unclear and likely to be critical to computations involved in navigation such as path integration. Here, we used 2-photon cellular imaging of neural activity of putative excitatory (CaMKII expressing) and inhibitory (parvalbumin expressing) neurons to measure visual and locomotion evoked activity in RSC and compare it to primary visual cortex (V1). We observed stimulus position and orientation tuning, and a retinotopic organization. Locomotion modulation of activity of single neurons, both in darkness and light, was more pronounced in RSC than V1, and while locomotion modulation was strongest in RSC parvalbumin-positive neurons, visual-locomotion integration was found to be more supralinear in CaMKII neurons. Longitudinal measurements showed that response properties were stably maintained over many weeks. These data provide evidence for stable representations of visual cues in RSC that are spatially selective. These may provide sensory data to contribute to the formation of memories of spatial information.

Mammillothalamic Disconnection Alters Hippocampocortical Oscillatory Activity and Microstructure: Implications for Diencephalic Amnesia.

Diencephalic amnesia can be as debilitating as the more commonly known temporal lobe amnesia, yet the precise contribution of diencephalic structures to memory processes remains elusive. Across four cohorts of male rats, we used discrete lesions of the mammillothalamic tract to model aspects of diencephalic amnesia and assessed the impact of these lesions on multiple measures of activity and plasticity within the hippocampus and retrosplenial cortex. Lesions of the mammillothalamic tract had widespread indirect effects on hippocampocortical oscillatory activity within both theta and gamma bands. Both within-region oscillatory activity and cross-regional synchrony were altered. The network changes were state-dependent, displaying different profiles during locomotion and paradoxical sleep. Consistent with the associations between oscillatory activity and plasticity, complementary analyses using several convergent approaches revealed microstructural changes, which appeared to reflect a suppression of learning-induced plasticity in lesioned animals. Together, these combined findings suggest a mechanism by which damage to the medial diencephalon can impact upon learning and memory processes, highlighting an important role for the mammillary bodies in the coordination of hippocampocortical activity.SIGNIFICANCE STATEMENT Information flow within the Papez circuit is critical to memory. Damage to ascending mammillothalamic projections has consistently been linked to amnesia in humans and spatial memory deficits in animal models. Here we report on the changes in hippocampocortical oscillatory dynamics that result from chronic lesions of the mammillothalamic tract and demonstrate, for the first time, that the mammillary bodies, independently of the supramammillary region, contribute to frequency modulation of hippocampocortical theta oscillations. Consistent with the associations between oscillatory activity and plasticity, the lesions also result in a suppression of learning-induced plasticity. Together, these data support new functional models whereby mammillary bodies are important for coordinating hippocampocortical activity rather than simply being a relay of hippocampal information as previously assumed.

Plasticity in Adult Mouse Visual Cortex Following Optic Nerve Injury.

Optic nerve (ON) injury is an established model of axonal injury which results in retrograde degeneration and death of retinal ganglion cells as well anterograde loss of transmission and Wallerian degeneration of the injured axons. While the local impact of ON crush has been extensively documented we know comparatively little about the functional changes that occur in higher visual structures such as primary visual cortex (V1). We explored the extent of adult cortical plasticity using ON crush in aged mice. V1 function of the contralateral hemisphere was assessed longitudinally by intrinsic signal imaging and 2-photon calcium imaging before and after ON crush. Functional imaging demonstrated an immediate shift in V1 ocular dominance towards the ipsilateral, intact eye, due to the expected almost complete loss of responses to contralateral eye stimulation. Surprisingly, within 2 weeks we observed a delayed increase in ipsilateral eye responses. Additionally, spontaneous activity in V1 was reduced, similar to the lesion projection zone after retinal lesions. The observed changes in V1 activity indicate that severe ON injury in adulthood evokes cortical plasticity not only cross-modally but also within the visual cortex; this plasticity may be best compared with that seen after retinal lesions.

Spatial memory deficits initiated by agroclavine injection or olfactory bulbectomy in rats are characterized by different levels of long-term potentiation expression in the hippocampus.

Aim: To clarify whether long-term potentiation (LTP) is the mechanism underpinning mnemonic processes. Mathrials and methods: We studied LTP in hippocampal slices from rats whose spatial memory deficit was produced by either olfactory bulbectomy (OBX) or pretreatment with an ergot alkaloid, agroclavine. OBX is accompanied by cholinergic system inhibition whereas agroclavine predominantly activates dopaminergic mediation. The both have been shown to be involved in learning/memory and LTP mechanisms.Results: In OBX- vs. sham-operated rat, we have revealed significant reduction of LTP in hippocampal CA1 region. In contrast, no LTP differences in agroclavine- vs. vehicle-treated rats were observed. Conclusions: These results demonstrate that LTP expression in the hippocampus is dependent on the origin of spatial memory impairment. Furthermore, they suggest that pharmacological and neurodegenerative models of AD might be useful approach for discovery of both AD mechanisms and mixed pathology dementias.

Animal models of amblyopia.

Unquestionably, the last six decades of research on various animal models have advanced our understanding of the mechanisms that underlie the many complex characteristics of amblyopia as well as provided promising new avenues for treatment. While animal models in general have served an important purpose, there nonetheless remain questions regarding the efficacy of particular models considering the differences across animal species, especially when the goal is to provide the foundations for human interventions. Our discussion of these issues culminated in three recommendations for future research to provide cohesion across animals models as well as a fourth recommendation for acceptance of a protocol for the minimum number of steps necessary for the translation of results obtained on particular animal models to human clinical trials. The three recommendations for future research arose from discussions of various issues including the specific results obtained from the use of different animal models, the degree of similarity to the human visual system, the ability to generate animal models of the different types of human amblyopia as well as the difficulty of scaling developmental timelines between different species.

Homeostatic plasticity mechanisms are required for juvenile, but not adult, ocular dominance plasticity.

Ocular dominance (OD) plasticity in the visual cortex is a classic model system for understanding developmental plasticity, but the visual cortex also shows plasticity in adulthood. Whether the plasticity mechanisms are similar or different at the two ages is not clear. Several plasticity mechanisms operate during development, including homeostatic plasticity, which acts to maintain the total excitatory drive to a neuron. In agreement with this idea, we found that an often-studied substrain of C57BL/6 mice, C57BL/6JOlaHsd (6JOla), lacks both the homeostatic component of OD plasticity as assessed by intrinsic signal imaging and synaptic scaling of mEPSC amplitudes after a short period of dark exposure during the critical period, whereas another substrain, C57BL/6J (6J), exhibits both plasticity processes. However, in adult mice, OD plasticity was identical in the 6JOla and 6J substrains, suggesting that adult plasticity occurs by a different mechanism. Consistent with this interpretation, adult OD plasticity was normal in TNFα knockout mice, which are known to lack juvenile synaptic scaling and the homeostatic component of OD plasticity, but was absent in adult α-calcium/calmodulin-dependent protein kinase II;T286A (αCaMKII(T286A)) mice, which have a point mutation that prevents autophosphorylation of αCaMKII. We conclude that increased responsiveness to open-eye stimulation after monocular deprivation during the critical period is a homeostatic process that depends mechanistically on synaptic scaling during the critical period, whereas in adult mice it is mediated by a different mechanism that requires αCaMKII autophosphorylation. Thus, our study reveals a transition between homeostatic and long-term potentiation-like plasticity mechanisms with increasing age.

The role of GluA1 in ocular dominance plasticity in the mouse visual cortex.

Ocular dominance plasticity is a widely studied model of experience-dependent cortical plasticity. It has been shown that potentiation of open eye responses resulting from monocular deprivation relies on a homeostatic response to loss of input from the closed eye, but the mechanisms by which this occurs are not fully understood. The role of GluA1 in the homeostatic component of ocular dominance (OD) plasticity has not so far been tested. In this study, we tested the idea that the GluA1 subunit of the AMPA receptor is necessary for open eye potentiation. We found that open eye potentiation did not occur in GluA1 knock-out (GluA1(-/-)) mice but did occur in wild-type littermates when monocular deprivation was imposed during the critical period. We also found that depression of the closed eye response that normally occurs in the monocular as well as binocular zone is delayed, but only in the monocular zone in GluA1(-/-) mice and only in a background strain we have previously shown lacks synaptic scaling (C57BL/6OlaHsd). In adult mice, we found that OD plasticity and facilitation of OD plasticity by prior monocular experience were both present in GluA1(-/-) mice, suggesting that the GluA1-dependent mechanisms only operate during the critical period.

Effects of digesting chondroitin sulfate proteoglycans on plasticity in cat primary visual cortex.

Monocular deprivation (MD) during a critical period of postnatal development produces significant changes in the anatomy and physiology of the visual cortex, and the deprived eye becomes amblyopic. Extracellular matrix molecules have a major role in restricting plasticity such that the ability to recover from MD decreases with age. Chondroitin sulfate proteoglycans (CSPGs) act as barriers to cell migration and axon growth. Previous studies showing that degradation of CSPGs by the bacterial enzyme chondroitinase can restore plasticity in the adult rat visual cortex suggest a potential treatment for amblyopia. Here MD was imposed in cats from the start of the critical period until 3.5 months of age. The deprived eye was reopened, the functional architecture of the visual cortex was assessed by optical imaging of intrinsic signals, and chondroitinase was injected into one hemisphere. Imaging was repeated 1 and 2 weeks postinjection, and visually evoked potentials (VEPs) and single-cell activity were recorded. Immunohistochemistry showed that digestion of CSPGs had been successful. After 2 weeks of binocular exposure, some recovery of deprived-eye responses occurred when chondroitinase had been injected into the hemisphere contralateral to that eye; when injected into the ipsilateral hemisphere, no recovery was seen. Deprived-eye VEPs were no larger in the injected hemisphere than in the opposite hemisphere. The small number of neurons dominated by the deprived eye exhibited poor tuning characteristics. These results suggest that despite structural effects of chondroitinase in adult cat V1, plasticity was not sufficiently restored to enable significant functional recovery of the deprived eye.

Stripe-rearing changes multiple aspects of the structure of primary visual cortex.

An important example of brain plasticity is the change in the structure of the orientation map in mammalian primary visual cortex in response to a visual environment consisting of stripes of one orientation. In principle there are many different ways in which the structure of a normal map could change to accommodate increased preference for one orientation. However, until now these changes have been characterised only by the relative sizes of the areas of primary visual cortex representing different orientations. Here we extend to the stripe-reared case a recently proposed Bayesian method for reconstructing orientation maps from intrinsic signal optical imaging data. We first formulated a suitable prior for the stripe-reared case, and developed an efficient method for maximising the marginal likelihood of the model in order to determine the optimal parameters. We then applied this to a set of orientation maps from normal and stripe-reared cats. This analysis revealed that several parameters of overall map structure, specifically the difference between wavelength, scaling and mean of the two vector components of maps, changed in response to stripe-rearing, which together give a more nuanced assessment of the effect of rearing condition on map structure than previous measures. Overall this work expands our understanding of the effects of the environment on brain structure.

A novel system for the classification of diseased retinal ganglion cells.

Retinal ganglion cell (RGC) dendritic atrophy is an early feature of many forms of retinal degeneration, providing a challenge to RGC classification. The characterization of these changes is complicated by the possibility that selective labeling of any particular class can confound the estimation of dendritic remodeling. To address this issue we have developed a novel, robust, and quantitative RGC classification based on proximal dendritic features which are resistant to early degeneration. RGCs were labeled through the ballistic delivery of DiO and DiI coated tungsten particles to whole retinal explants of 20 adult Brown Norway rats. RGCs were grouped according to the Sun classification system. A comprehensive set of primary and secondary dendrite features were quantified and a new classification model derived using principal component (PCA) and discriminant analyses, to estimate the likelihood that a cell belonged to any given class. One-hundred and thirty one imaged RGCs were analyzed; according to the Sun classification, 24% (n = 31) were RGCA, 29% (n = 38) RGCB, 32% (n = 42) RGCC, and 15% (n = 20) RGCD. PCA gave a 3 component solution, separating RGCs based on descriptors of soma size and primary dendrite thickness, proximal dendritic field size and dendritic tree asymmetry. The new variables correctly classified 73.3% (n = 74) of RGCs from a training sample and 63.3% (n = 19) from a hold out sample indicating an effective model. Soma and proximal dendritic tree morphological features provide a useful surrogate measurement for the classification of RGCs in disease. While a definitive classification is not possible in every case, the technique provides a useful safeguard against sample bias where the normal criteria for cell classification may not be reliable.

Experience-dependent regulation of functional maps and synaptic protein expression in the cat visual cortex.

Although the basis of our knowledge of experience-dependent plasticity comes from studies on carnivores and primates, studies examining the physiological and molecular mechanisms that underlie development and plasticity have increasingly employed mice. We have used several common rearing paradigms, such as dark-rearing and monocular deprivation (MD), to examine the timing of the physiological and molecular changes to altered experience in the cat primary visual cortex. Dark-rearing from birth or for 1 week starting at 4 weeks of age produced a similar reduction in the amplitude of responses measured through intrinsic signal imaging and a reduction in orientation selectivity. One week of visual experience following dark-rearing until 4 weeks of age yielded normal responses in both amplitude and orientation selectivity. The depression of deprived-eye responses was similar in magnitude after 2 and 7 days of MD. In contrast, non-deprived-eye responses almost doubled in magnitude after 7 days compared with 2 days of MD. These changes in the functional properties of primary visual cortex neurons were mirrored by specific changes in synaptic protein expression. Changes in proteins such as the NR2A and NR2B subunits of the N-methyl-D-aspartate receptor, postsynaptic density protein 95, alpha-CA(2+) /calmodulin-dependent protein kinase II (αCaMKII), and GABA(A) α1a indicated that the levels of sensory activity regulated mechanisms associated with both excitatory (NR2A and NR2B) and inhibitory (GABA(A) α1a) transmission so as to maintain response homeostasis. Additionally, we found that MD regulated the AMPA receptor glutamate (GluR1) subunit as well as signalling molecules (αCaMKII and synaptic Ras GTPase activating protein, SynGAP) downstream of N-methyl-D-aspartate receptors. Proteins in a common signalling pathway appeared to have similar developmental expression profiles that were broadly similar between cats and rodents.

Effects of different forms of monocular deprivation on primary visual cortex maps.

Monocular deprivation (MD) by lid suture is one of the classic paradigms for the study of developmental plasticity in the cerebral cortex, and we have detailed knowledge of its anatomical and physiological consequences as well as underlying molecular and cellular mechanisms. However, the effects of other forms of manipulating visual input through one eye on the functional architecture of the primary visual cortex (V1) have not yet been examined directly. We compared MD by lid suture with the effects of daily monocular lens wear using either a frosted lens or a neutral density (ND) filter. We used optical imaging of intrinsic signals and visually evoked potentials (VEPs) to assess responses in V1 to monocular stimulation. We found that loss of stimulus contrast through monocular frosted lens wear resulted in marked takeover of cortical territory by the nondeprived eye (NDE) similar to that caused by classic MD, and in virtual absence of orientation-selective responses following stimulation of the deprived eye (DE). Furthermore, amplitudes of VEPs in response to gratings of a range of spatial frequencies were significantly reduced in the DE compared to the NDE. In contrast, differences in luminance between two eyes caused by an ND filter in front of one eye did not affect ocular dominance and orientation maps, and there was no significant difference in the amplitude of VEPs elicited through the two eyes. Our results are consistent with previous electrophysiological studies in demonstrating that binocular pattern information is necessary to maintain normal functional maps in both eyes, while reduced luminance in one eye has little effect on the overall functional architecture and visual responses in V1.

Recruitment of frontal sensory circuits during visual discrimination.

A long-range circuit linking the medial frontal cortex to the primary visual cortex (V1) has been proposed to mediate visual selective attention in mice during visually guided behavior. Here, we use in vivo two-photon functional imaging to measure the endogenous activity of axons of A24b/M2 neurons from this region projecting to layer 1 of V1 (A24b/M2-V1axons) in mice either passively viewing stimuli or performing a go/no-go visually guided task. We observe that while A24b/M2-V1axons are recruited under these conditions, this is not linked to enhancement of neural or behavioral measures of sensory coding. Instead, A24b/M2-V1axon activity is associated with licking behavior, modulated by reward, and biased toward the sensory cortical hemisphere representing the stimulus currently being discriminated.

Refinements to rodent head fixation and fluid/food control for neuroscience.

The use of head fixation in mice is increasingly common in research, its use having initially been restricted to the field of sensory neuroscience. Head restraint has often been combined with fluid control, rather than food restriction, to motivate behaviour, but this too is now in use for both restrained and non-restrained animals. Despite this, there is little guidance on how best to employ these techniques to optimise both scientific outcomes and animal welfare. This article summarises current practices and provides recommendations to improve animal wellbeing and data quality, based on a survey of the community, literature reviews, and the expert opinion and practical experience of an international working group convened by the UK's National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs). Topics covered include head fixation surgery and post-operative care, habituation to restraint, and the use of fluid/food control to motivate performance. We also discuss some recent developments that may offer alternative ways to collect data from large numbers of behavioural trials without the need for restraint. The aim is to provide support for researchers at all levels, animal care staff, and ethics committees to refine procedures and practices in line with the refinement principle of the 3Rs.

Binocular vision: only half a brain needed.

A recent study on zebrafish has shown that, by rerouting afferents from two eyes into a normally monocular brain structure, a fully functional binocular circuitry can be made to develop spontaneously.

Protection against deprivation amblyopia depends on relative not absolute daily binocular exposure.

Short daily periods of binocular exposure (BE) can offset longer single daily episodes of monocular exposure (ME) to prevent the development of deprivation amblyopia. To determine whether the outcome depended upon an absolute daily amount of BE or its proportion of the daily visual exposure, daily mixed visual input of 3 different durations (3.5, 7, or 12 h) was imposed on 3 cohorts of kittens. Measurements of the visual acuity of the deprived eye at the end of mixed daily visual input revealed that the acuity of the deprived eye developed to normal values so long as the proportion of the total exposure that was binocular was 30% or more. By contrast, the development of functional ocular dominance domains in V1 revealed by optical imaging suggests that normal domains emerge with a fixed amount of daily binocular exposure. The latter result is consistent with the effects of any daily period of ME, or BE, or both, effectively saturating with a small dose so that the effects of ME of any length can be offset by a short period of BE. The different result for vision may reflect neural events at higher and/or multiple levels in the visual pathway.

Contrast adaptation and interocular transfer in cortical cells: A re-analysis & a two-stage gain-control model of binocular combination.

How do V1 cells respond to, adapt to, and combine signals from the two eyes? We tested a simple functional model that has monocular and binocular stages of divisive contrast gain control (CGC) that sit before, and after, binocular summation respectively. Interocular suppression (IOS) was another potential influence on contrast gain. Howarth, Vorobyov & Sengpiel (2009, Cerebral Cortex, 19, 1835-1843) studied contrast adaptation and interocular transfer in cat V1 cells. In our re-analysis we found that ocular dominance (OD) and contrast adaptation at a fixed test contrast were well described by a re-scaling of the unadapted orientation tuning curve - a simple change in response gain. We compared six variants of the basic model, and one model fitted the gain data notably better than the others did. When the dominant eye was tested, adaptation reduced cell response gain more when that eye was adapted than when the other eye was adapted. But when the non-dominant eye was tested, adapting either eye gave about the same reduction in overall gain, and there was an interaction between OD and adapting eye that was well described by the best-fitting model. Two key features of this model are that signals driving IOS arise 'early', before attenuation due to OD, while suppressive CGC signals are 'late' and so affected by OD. We show that late CGC confers a functional advantage: it yields partial compensation for OD, which should reduce ocular imbalance at the input to binocular summation, and improve the cell's sensitivity to variation in stereo disparity.