Cortical Inactivation Research Articles

Motor cortical inactivation impairs corrective submovements in mice performing a hold-still center-out reach task.

Holding still and aiming reaches to spatial targets may depend on distinct neural circuits. Using automated homecage training and a sensitive joystick, we trained freely moving mice to contact a joystick, hold their forelimb still, and then reach to rewarded target locations. Mice learned the task by initiating forelimb sequences with clearly resolved submillimeter-scale micromovements followed by millimeter-scale reaches to learned spatial targets. Hundreds of thousands of trajectories were decomposed into millions of kinematic submovements, while photoinhibition was used to test roles of motor cortical areas. Inactivation of both caudal and rostral forelimb areas preserved the ability to produce aimed reaches, but reduced reach speed. Inactivation specifically of contralateral caudal forelimb area (CFA) additionally impaired the ability to aim corrective submovements to remembered locations following target undershoots. Our findings show that motor cortical inactivations reduce the gain of forelimb movements but that inactivation specifically of contralateral CFA impairs corrective movements important for reaching a target location.NEW & NOTEWORTHY To test the role of different cortical areas in holding still and reaching to targets, this study combined home-cage training with optogenetic silencing as mice engaged in a learned center-out-reach task. Inactivation specifically of contralateral caudal forelimb area (CFA) impaired corrective movements necessary to reach spatial targets to earn reward.

Effects of unilateral cortical inactivation on monoamine metabolism in symmetrical forebrain areas of white outbred mice

BACKGROUND: Clinical observations and animal experiments have shown the lateral specificity of the action of compounds that alter monoaminergic transmission. However, the mechanism of this phenomenon has not yet been studied.AIM:To evaluate the effect of unilateral cortical spreading depression on monoamine metabolism in white outbred mice. MATERIALS AND METHODS:Eighteen sexually mature male white outbred mice were studied. Functional inactivation of the cortex of one of the cerebral hemispheres was induced using unilateral epidural application of 1 × 1 mm filter paper moistened with 25% KCl solution. Then, 15 minutes after exposure, the animals were decapitated. The norepinephrine, dopamine, serotonin, and metabolite (i.e., dioxyphenylacetic (DOPAC), homovanilinic (HVA), and 5-hydroxyindolacetic (5-HIAA) acids) content of the cerebral cortex, hippocampus, olfactory tubercle, and striatum was measured using the HPLC method with an electrochemical detector. RESULTS: The inactivation of the left hemisphere was due to a bilateral decrease in norepinephrine in the hippocampus, an ipsilateral increase in extracellular metabolism of dopamine (HVA and/or HVA/dopamine) in the olfactory tubercle and striatum, and a contralateral increase of dopamine in the cortex and hippocampus. Furthermore, the DOPAC/dopamine ratio in the right olfactory tubercle decreased with left hemisphere inactivation. Right hemisphere inactivation did not cause bilateral effects. With inactivation of the right hemisphere on the inactivation side, norepinephrine in the hippocampus decreased and HVA in the striatum increased. In the left side, with the inactivation of the right hemisphere mice, dopamine in the hippocampus increased, as well as the DOPAC level and DOPAC/dopamine ratio in the olfactory tubercle. Serotonin metabolism (5-HIAA/serotonin) in the right hippocampus and left olfactory tubercle increased only at inactivation of the right hemisphere.CONCLUSIONS:The monoaminergic effects of functional inactivation of the left and right hemisphere cortex in white outbred mice are not mirror-symmetrical.

Reversible Inactivation of Ferret Auditory Cortex Impairs Spatial and Nonspatial Hearing.

A key question in auditory neuroscience is to what extent are brain regions functionally specialized for processing specific sound features, such as location and identity. In auditory cortex, correlations between neural activity and sounds support both the specialization of distinct cortical subfields, and encoding of multiple sound features within individual cortical areas. However, few studies have tested the contribution of auditory cortex to hearing in multiple contexts. Here we determined the role of ferret primary auditory cortex in both spatial and nonspatial hearing by reversibly inactivating the middle ectosylvian gyrus during behavior using cooling (n = 2 females) or optogenetics (n = 1 female). Optogenetic experiments used the mDLx promoter to express Channelrhodopsin-2 in GABAergic interneurons, and we confirmed both viral expression (n = 2 females) and light-driven suppression of spiking activity in auditory cortex, recorded using Neuropixels under anesthesia (n = 465 units from 2 additional untrained female ferrets). Cortical inactivation via cooling or optogenetics impaired vowel discrimination in colocated noise. Ferrets implanted with cooling loops were tested in additional conditions that revealed no deficit when identifying vowels in clean conditions, or when the temporally coincident vowel and noise were spatially separated by 180 degrees. These animals did, however, show impaired sound localization when inactivating the same auditory cortical region implicated in vowel discrimination in noise. Our results demonstrate that, as a brain region showing mixed selectivity for spatial and nonspatial features of sound, primary auditory cortex contributes to multiple forms of hearing.SIGNIFICANCE STATEMENT Neurons in primary auditory cortex are often sensitive to the location and identity of sounds. Here we inactivated auditory cortex during spatial and nonspatial listening tasks using cooling, or optogenetics. Auditory cortical inactivation impaired multiple behaviors, demonstrating a role in both the analysis of sound location and identity and confirming a functional contribution of mixed selectivity observed in neural activity. Parallel optogenetic experiments in two additional untrained ferrets linked behavior to physiology by demonstrating that expression of Channelrhodopsin-2 permitted rapid light-driven suppression of auditory cortical activity recorded under anesthesia.

Increased fMRI connectivity upon chemogenetic inhibition of the mouse prefrontal cortex

While shaped and constrained by axonal connections, fMRI-based functional connectivity reorganizes in response to varying interareal input or pathological perturbations. However, the causal contribution of regional brain activity to whole-brain fMRI network organization remains unclear. Here we combine neural manipulations, resting-state fMRI and in vivo electrophysiology to probe how inactivation of a cortical node causally affects brain-wide fMRI coupling in the mouse. We find that chronic inhibition of the medial prefrontal cortex (PFC) via overexpression of a potassium channel increases fMRI connectivity between the inhibited area and its direct thalamo-cortical targets. Acute chemogenetic inhibition of the PFC produces analogous patterns of fMRI overconnectivity. Using in vivo electrophysiology, we find that chemogenetic inhibition of the PFC enhances low frequency (0.1–4 Hz) oscillatory power via suppression of neural firing not phase-locked to slow rhythms, resulting in increased slow and δ band coherence between areas that exhibit fMRI overconnectivity. These results provide causal evidence that cortical inactivation can counterintuitively increase fMRI connectivity via enhanced, less-localized slow oscillatory processes.

Contributions of Distinct Auditory Cortical Inhibitory Neuron Types to the Detection of Sounds in Background Noise.

The ability to separate background noise from relevant acoustic signals is essential for appropriate sound-driven behavior in natural environments. Examples of this separation are apparent in the auditory system, where neural responses to behaviorally relevant stimuli become increasingly noise invariant along the ascending auditory pathway. However, the mechanisms that underlie this reduction in responses to background noise are not well understood. To address this gap in knowledge, we first evaluated the effects of auditory cortical inactivation on mice of both sexes trained to perform a simple auditory signal-in-noise detection task and found that outputs from the auditory cortex are important for the detection of auditory stimuli in noisy environments. Next, we evaluated the contributions of the two most common cortical inhibitory cell types, parvalbumin-expressing (PV+) and somatostatin-expressing (SOM+) interneurons, to the perception of masked auditory stimuli. We found that inactivation of either PV+ or SOM+ cells resulted in a reduction in the ability of mice to determine the presence of auditory stimuli masked by noise. These results indicate that a disruption of auditory cortical network dynamics by either of these two types of inhibitory cells is sufficient to impair the ability to separate acoustic signals from noise.

Targeting the motor cortex to restore walking after incomplete spinal cord injury.

Targeting the motor cortex to restore walking after incomplete spinal cord injury.

Heterogeneous side effects of cortical inactivation in behaving animals.

Cortical inactivation represents a key causal manipulation allowing the study of cortical circuits and their impact on behavior. A key assumption in inactivation studies is that the neurons in the target area become silent while the surrounding cortical tissue is only negligibly impacted. However, individual neurons are embedded in complex local circuits composed of excitatory and inhibitory cells with connections extending hundreds of microns. This raises the possibility that silencing one part of the network could induce complex, unpredictable activity changes in neurons outside the targeted inactivation zone. These off-target side effects can potentially complicate interpretations of inactivation manipulations, especially when they are related to changes in behavior. Here, we demonstrate that optogenetic inactivation of glutamatergic neurons in the superficial layers of monkey primary visual cortex (V1) induces robust suppression at the light-targeted site, but destabilizes stimulus responses in the neighboring, untargeted network. We identified four types of stimulus-evoked neuronal responses within a cortical column, ranging from full suppression to facilitation, and a mixture of both. Mixed responses were most prominent in middle and deep cortical layers. These results demonstrate that response modulation driven by lateral network connectivity is diversely implemented throughout a cortical column. Importantly, consistent behavioral changes induced by optogenetic inactivation were only achieved when cumulative network activity was homogeneously suppressed. Therefore, careful consideration of the full range of network changes outside the inactivated cortical region is required, as heterogeneous side effects can confound interpretation of inactivation experiments.

Chronic inactivation of the contralesional hindlimb motor cortex after thoracic spinal cord hemisection impedes locomotor recovery in the rat

After incomplete spinal cord injury (SCI), cortical plasticity is involved in hindlimb locomotor recovery. Nevertheless, whether cortical activity is required for motor map plasticity and recovery remains unresolved. Here, we combined a unilateral thoracic spinal cord injury (SCI) with a cortical inactivation protocol that uncovered a functional role of contralesional cortical activity in hindlimb recovery and ipsilesional map plasticity. In adult rats, left hindlimb paralysis was induced by sectioning half of the spinal cord at the thoracic level (hemisection) and we used a continuous infusion of muscimol (GABAA agonist, 10 mM, 0.11 µl/h) delivered via implanted osmotic pump (n = 9) to chronically inactivate the contralesional hindlimb motor cortex. Hemisected rats with saline infusion served as a SCI control group (n = 8), and intact rats with muscimol infusion served as an inactivation control group (n = 6). Locomotion was assessed in an open field, on a horizontal ladder, and on a treadmill prior to and for three weeks after hemisection. Cortical inactivation after hemisection significantly impeded hindlimb locomotor recovery in all tasks and specifically disrupted the ability of rats to generate proper flexion of the affected hindlimb during stepping compared to SCI controls, with no significant effect of inactivation in intact rats. Chronic and acute (n = 4) cortical inactivation after hemisection also significantly reduced the representation of the affected hindlimb in the ipsilesional motor cortex derived with intracortical microsimulation (ICMS). Our results provide evidence that residual activity in the contralesional hindlimb motor cortex after thoracic hemisection contributes to spontaneous locomotor recovery and map plasticity.

Constraint-induced intervention as an emergent phenomenon from synaptic competition in biological systems

The principle of constraint-induced therapy is widely practiced in rehabilitation. In hemiplegic cerebral palsy (CP) with impaired contralateral corticospinal projection due to unilateral injury, function improves after imposing a temporary constraint on limbs from the less affected hemisphere. This type of partially-reversible impairment in motor control by early brain injury bears a resemblance to the experience-dependent plastic acquisition and modification of neuronal response selectivity in the visual cortex. Previously, such mechanism was modeled within the framework of BCM (Bienenstock-Cooper-Munro) theory, a rate-based synaptic modification theory. Here, we demonstrate a minimally complex yet sufficient neural network model which provides a fundamental explanation for inter-hemispheric competition using a simplified spike-based model of information transmission and plasticity. We emulate the restoration of function in hemiplegic CP by simulating the competition between cells of the ipsilateral and contralateral corticospinal tracts. We use a high-speed hardware neural simulation to provide realistic numbers of spikes and realistic magnitudes of synaptic modification. We demonstrate that the phenomenon of constraint-induced partial reversal of hemiplegia can be modeled by simplified neural descending tracts with 2 layers of spiking neurons and synapses with spike-timing-dependent plasticity (STDP). We further demonstrate that persistent hemiplegia following unilateral cortical inactivation or deprivation is predicted by the STDP-based model but is inconsistent with BCM model. Although our model is a highly simplified and limited representation of the corticospinal system, it offers an explanation of how constraint as an intervention can help the system to escape from a suboptimal solution. This is a display of an emergent phenomenon from the synaptic competition.

Cortical Inactivation Does Not Block Response Enhancement in the Superior Colliculus.

Repetitive visual stimulation is successfully used in a study on the visual evoked potential (VEP) plasticity in the visual system in mammals. Practicing visual tasks or repeated exposure to sensory stimuli can induce neuronal network changes in the cortical circuits and improve the perception of these stimuli. However, little is known about the effect of visual training at the subcortical level. In the present study, we extend the knowledge showing positive results of this training in the rat’s Superior colliculus (SC). In electrophysiological experiments, we showed that a single training session lasting several hours induces a response enhancement both in the primary visual cortex (V1) and in the SC. Further, we tested if collicular responses will be enhanced without V1 input. For this reason, we inactivated the V1 by applying xylocaine solution onto the cortical surface during visual training. Our results revealed that SC’s response enhancement was present even without V1 inputs and showed no difference in amplitude comparing to VEPs enhancement while the V1 was active. These data suggest that the visual system plasticity and facilitation can develop independently but simultaneously in different parts of the visual system.

Mapping Functional Connectivity from the Dorsal Cortex to the Thalamus

Neural activity in the corticothalamic network is crucial for sensation, memory, decision, and action. Nevertheless, a systematic characterization of corticothalamic functional connectivity has not been achieved. Here, we developed a high throughput method to systematically map functional connections from the dorsal cortex to the thalamus in awake mice by combing optogenetic inactivation with multi-channel recording. Cortical inactivation resulted in a rapid reduction of thalamic activity, revealing topographically organized corticothalamic excitatory inputs. Cluster analysis showed that groups of neurons within individual thalamic nuclei exhibited distinct dynamics. The effects of inactivation evolved with time and were modulated by behavioral states. Furthermore, we found that a subset of thalamic neurons received convergent inputs from widespread cortical regions. Our results present a framework for collecting, analyzing, and presenting large electrophysiological datasets with region-specific optogenetic perturbations and serve as a foundation for further investigation of information processing in the corticothalamic pathway.

Medial Prefrontal Cortical Modulation of Whisker Thalamic Responses in Anesthetized Rats

The medial prefrontal cortex (mPFC) has been implicated in novelty detection and attention. We studied the effect of mPFC electrical stimulation on whisker responses recorded in the ventroposterior medial thalamic nucleus (VPM), the posterior thalamic nucleus (POm) and the primary somatosensory (S1) cortex in urethane anesthetized rats. Field potentials and unit recordings were performed in the VPM or POm thalamic nuclei, in S1 cortex, and in the Zona Incerta (ZI). Somatosensory evoked potentials were elicited by whisker deflections. Current pulses were delivered by bipolar stimulating electrodes aimed at the prelimbic (PL) or infralimbic (IL) areas of mPFC. PL train stimulation (50 Hz, 500 ms) induced a facilitation of whisker responses in the VPM nucleus that lasted minutes and a short inhibition in the POm nucleus. IL stimulation induced a facilitation of whisker responses in both VPM and POm nuclei. Facilitation was due to corticofugal projections because it was reduced after S1 cortical inactivation with lidocaine, and by activation of NMDA glutamatergic receptors because it was blocked by APV. Paired stimulation of mPFC and whiskers revealed an inhibitory effect at short intervals (<100 ms), which was mediated by ZI inhibitory neurons since PL stimulation induced response facilitation in the majority of ZI neurons (42%) and muscimol injection into ZI nucleus reduced inhibitory effects, suggesting that the mPFC may inhibit the POm neurons by activation of GABAergic ZI neurons. In conclusion, the mPFC may control the flow of somatosensory information through the thalamus by activation of S1 and ZI neurons.

Ipsilesional Motor Cortex Plasticity Participates in Spontaneous Hindlimb Recovery after Lateral Hemisection of the Thoracic Spinal Cord in the Rat.

After an incomplete spinal cord injury (SCI) spontaneous motor recovery can occur in mammals, but the underlying neural substrates remain poorly understood. The motor cortex is crucial for skilled motor learning and the voluntary control of movement and is known to reorganize after cortical injury to promote recovery. Motor cortex plasticity has also been shown to parallel the recovery of forelimb function after cervical SCI, but whether cortical plasticity participates in hindlimb recovery after SCI remains unresolved. Using intracortical microstimulation (ICMS) mapping, behavioral and cortical inactivation techniques in the female Long-Evans rat, we evaluated the spontaneous cortical mechanisms of hindlimb motor recovery 1-5 weeks after lateral hemisection of the thoracic (T8) spinal cord that ablated the crossed corticospinal tract (CST) from the contralesional motor cortex while sparing the majority of the CST from the ipsilesional motor cortex. Hemisection initially impaired hindlimb motor function bilaterally but significant recovery occurred during the first 3 weeks. ICMS revealed time-dependent changes in motor cortex organization, characterized by a chronic abolishment of hindlimb motor representation in the contralesional motor cortex and the development of transient bilateral hindlimb representation in the ipsilesional motor cortex 3 weeks after hemisection, when significant behavioral recovery occurred. Consistently, reversible inactivation of the ipsilesional, but not the contralesional motor cortex, during skilled ladder walking 3 weeks after hemisection reinstated deficits in both hindlimbs. These findings indicate that the ipsilesional motor cortex transiently reorganizes after lateral hemisection of the thoracic spinal cord to support recovery of hindlimb motor function.SIGNIFICANCE STATEMENT Partial motor recovery can occur after an incomplete spinal cord injury and is hypothesized to result from the reorganization of spared descending motor pathways. The motor cortex is crucial for the control of voluntary movement and contains topographical movement representations (motor maps) that are highly plastic. We examined the organization of hindlimb motor maps bilaterally after a lateral hemisection of the spinal cord to show that while motor maps are abolished in the deefferented cortex, the spared ipsilesional cortex transiently reorganizes to gain a representation of the affected hindlimb after injury that relates to recovery. This finding demonstrates that plasticity in the ipsilesional motor cortex at early time points after spinal cord hemisection is initially important to support motor recovery.

The effects of amygdala and cortical inactivation on taste neophobia.

The current study examined the effects of transient inactivation of the basolateral amygdala (BLA; Experiment 1) and gustatory cortex (GC; Experiment 2) on the expression of taste neophobia and its recovery. We found that inactivation (induced by infusions of baclofen/muscimol) of each structure before exposure to a novel saccharin (0.5%) solution elevated intake on Trial 1 (i.e., taste neophobia was attenuated) and, surprisingly, decreased intake on Trial 2. It seems unlikely that this intake reduction on Trial 2 can be attributed to taste aversion learning caused by drug infusions because in the subsequent experiments with the same set of the implanted animals, the rats did not decrease intake when baclofen/muscimol was infused after taste presentation on Trial 1. The latter result suggests that BLA or GC inactivation that attenuates taste neophobia may also impair memory consolidation of a safe taste experience.

Lateral geniculate neurons projecting to primary visual cortex show ocular dominance plasticity in adult mice.

Experience-dependent plasticity in the mature visual system is widely considered to be cortical. Using chronic two-photon Ca2+ imaging of thalamic afferents in layer 1 of binocular visual cortex, we provide evidence against this tenet: the respective dorsal lateral geniculate nucleus (dLGN) cells showed pronounced ocular dominance (OD) shifts after monocular deprivation in adult mice. Most (86%), but not all, of dLGN cell boutons were monocular during normal visual experience. Following deprivation, initially deprived-eye-dominated boutons reduced or lost their visual responsiveness to that eye and frequently became responsive to the non-deprived eye. This cannot be explained by eye-specific cortical changes propagating to dLGN via cortico-thalamic feedback because the shift in dLGN responses was largely resistant to cortical inactivation using the GABAA receptor agonist muscimol. Our data suggest that OD shifts observed in the binocular visual cortex of adult mice may at least partially reflect plasticity of eye-specific inputs onto dLGN neurons.

Inactivation of Cerebellar Cortical Crus II Disrupts Temporal Processing of Absolute Timing but not Relative Timing in Voluntary Movements.

Several recent studies have demonstrated that the cerebellum plays an important role in temporal processing at the scale of milliseconds. However, it is not clear whether intrinsic cerebellar function involves the temporal processing of discrete or continuous events. Temporal processing during discrete events functions by counting absolute time like a stopwatch, while during continuous events it measures events at intervals. During the temporal processing of continuous events, animals might respond to rhythmic timing of sequential responses rather than to the absolute durations of intervals. Here, we tested the contribution of the cerebellar cortex to temporal processing of absolute and relative timings in voluntary movements. We injected muscimol and baclofen to a part of the cerebellar cortex of rats. We then tested the accuracy of their absolute or relative timing prediction using two timing tasks requiring almost identical reaching movements. Inactivation of the cerebellar cortex disrupted accurate temporal prediction in the absolute timing task. The rats formed two groups based on the changes to their timing accuracy following one of two distinct patterns which can be described as longer or shorter declines in the accuracy of learned intervals. However, a part of the cerebellar cortical inactivation did not affect the rats’ performance of relative timing tasks. We concluded that a part of the cerebellar cortex, Crus II, contributes to the accurate temporal prediction of absolute timing and that the entire cerebellar cortex may be unnecessary in cases in which accurately knowing the absolute duration of an interval is not required for temporal prediction.

Abstract WP279: Disruption of Cortico-hippocampal Functional Connectivity Underlies the Memory Impairment Following Experimental Stroke

Introduction: The cognitive consequences and the underlying mechanisms leading to cognitive impairments after cerebrovascular occlusive diseases are still unclear. Previously we have shown that distal middle cerebral artery occlusion (dMCAO) led to hippocampal hypofunction and spatial memory impairment in the absence of direct hippocampal injury. Hypothesis: In light of the importance of interactions between the cortex and hippocampus during memory processing, we hypothesize that cortical stroke disrupts afferent excitatory input from the lesioned cortical area to other remote brain regions such as the hippocampus, which can be mimicked by pharmacological inactivation of the cortex. Methods: Adult male rats were subjected to either dMCAO or cortical inactivation with AMPA receptor antagonist CNQX, compared to controls by sham-operation or injection with artificial CSF, respectively. Hippocampal function was determined by the social transmission of food preference (STFP) test, activity mapping by Fos imaging and in vivo electrophysiology by multi-channel extracellular recording. Results: dMCAO and cortical inactivation both induced impaired memory performance as measured in the STFP task and reduced hippocampal activation. The memory impairment was not attributed to change in olfaction by stroke or inactivation, since it was not detected in either experimental group. An increase in the bursts of sharp-wave associated ripples (SPW-Rs) was detected during reperfusion after stroke and after CNQX injection. Further, cortical inactivation also induced a shift of the theta phase, suggesting an alteration in the dynamics of hippocampal-cortical interactions. Conclusions: Given the crucial role of hippocampal theta and SPW-Rs in memory function, our results suggest that cortical stroke leads to memory impairment by disrupting the functional connectivity between the cortex and hippocampus, albeit without causing direct injury to the latter.

Cooling of the auditory cortex modifies neuronal activity in the inferior colliculus in rats

There are powerful pathways descending from the auditory cortex (AC) to the inferior colliculus (IC), yet their function is not fully understood. The aim of this study is to examine the effects of a reversible cortical inactivation, achieved by cooling of the AC, on the responses of neurons in the rat IC. Extracellular single-unit or multi-unit activity was recorded in the IC of anaesthetized rats with a 16-channel multielectrode probe introduced along the IC dorso-ventral axis through the dorsal cortex (DCIC) to the central nucleus of the IC (CIC). Cooling of the AC produced an increase in spontaneous activity and magnitude of the sound-evoked response in 47% of the IC neurons. Maximal changes in the neuronal activity were observed in the DCIC and the central part of the CIC. The final segments of the sustained responses to 60 ms stimuli and the off responses were more affected than the onset segments. Inactivation of the AC resulted in a suppression of the post-excitatory inhibition and neuronal adaptation, which was reflected in a pronounced enhancement of synchronized responses to a series of fast repeated clicks. The response parameters recovered, at least partly, to the pre-cooling levels 1 h after the cooling cessation. The frequency tuning properties of the IC neurons did not show any significant changes during the cooling period. The results demonstrate that AC cooling inactivates excitatory corticofugal pathways and results in a less activated intrinsic inhibitory network in the IC.

Tactile response adaptation to whisker stimulation in the lemniscal somatosensory pathway of rats

Response adaptation is associated with attenuation of neural responses as the result of different mechanisms. However, the main function of adaptation may be to enhance the flow of relevant information transmission in sensory pathways. To study tactile response adaptation in the somatosensory pathway, unit recordings were performed in the principal trigeminal nucleus, ventro postero-medial thalamic nucleus and barrel cortex by means of tungsten microelectrodes in urethane anesthetized rats. Tactile stimuli consisted in 20ms duration whisker deflections at different frequencies (0.5–10Hz). Presumably pyramidal cortical neurons showed response adaptation at frequencies >2Hz while putative inhibitory cortical neurons did not show response adaptation at 0.5, 5 or 10Hz. Inhibitory activity was increased by muscimol application into the cortex (8mM, 0.1µl); in this condition cortical adaptation was not affected, suggesting that adaptation was not due to an increase of inhibitory mechanisms. Adaptation was also observed in subcortical structures although the response attenuation was lesser than in the barrel cortex. Adaptation remained in subcortical structures after reversible cortical inactivation by cooling the barrel cortex. Acetylcholine application (10μM; 0.1μl) into the barrel cortex reduced response adaptation through the activation of muscarinic receptors because the effect was blocked by intraperitoneal injection of atropine (1mg/kg), suggesting that adaptation may change according to the cortical Ach level. Results indicate that response adaptation increases along the somatosensory pathway probably to alter the sensitivity of neurons in order to encode sensory stimuli more efficiently and to enhance the detectability of rare stimuli.

Genetic evidence that Celsr3 and Celsr2, together with Fzd3, regulate forebrain wiring in a Vangl-independent manner.

Celsr3 and Fzd3, members of "core planar cell polarity" (PCP) genes, were shown previously to control forebrain axon guidance and wiring by acting in axons and/or guidepost cells. Here, we show that Celsr2 acts redundantly with Celsr3, and that their combined mutation mimics that of Fzd3. The phenotypes generated upon inactivation of Fzd3 in different forebrain compartments are similar to those in conditional Celsr2-3 mutants, indicating that Fzd3 and Celsr2-3 act in the same population of cells. Inactivation of Celsr2-3 or Fzd3 in thalamus does not affect forebrain wiring, and joint inactivation in cortex and thalamus adds little to cortical inactivation alone in terms of thalamocortical projections. On the other hand, joint inactivation perturbs strongly the formation of the barrel field, which is unaffected upon single cortical or thalamic inactivation, indicating a role for interactions between thalamic axons and cortical neurons in cortical arealization. Unexpectedly, forebrain wiring is normal in mice defective in Vangl1 and Vangl2, showing that, contrary to epithelial PCP, axon guidance can be Vangl independent in some contexts. Our results suggest that Celsr2-3 and Fzd3 regulate axonal navigation in the forebrain by using mechanisms different from classical epithelial PCP, and require interacting partners other than Vangl1-2 that remain to be identified.