Protocol for stimulating specific rodent limb receptive fields while recording in vivo somatosensory-evoked activity.

Studies on sensory information processing typically focus on whisker-related tactile information, overlooking the question of how sensory inputs from other body areas are processed at cortical levels. Here, we present a protocol for stimulating specific rodent limb receptive fields while recording in vivo somatosensory-evoked activity. We describe steps for localizing cortical-hindlimb coordinates using acute peripheral stimulation, electrode placement, and the application of electrical stimulation. This protocol overcomes the challenge of inducing a reproducible and consistent stimulation of specific limbs. For complete details on the use and execution of this protocol, please refer to Miguel-Quesada et al.1.

Increased excitability and reduced GABAergic levels in somatosensory cortex under chronic spinal cord injury.

The complete or partial damage of ascending somatosensory pathways produced by a spinal cord injury triggers changes in the somatosensory cortex consisting in a functional expansion of activity from intact cortical regions towards deafferented ones, a process known as cortical reorganization. However, it is still unclear whether cortical reorganization depends on the severity of the spinal cord damage or if a spinal cord injury always leads to a similar cortical reorganization process in the somatosensory cortex. To answer these open questions in the field, we obtained longitudinal somatosensory evoked responses from bilateral hindlimb and forelimb cortex from animals with chronic full-transection or contusive spinal cord injury at thoracic level (T9-T10) to induce sensory deprivation of hindlimb cortex while preserving intact the forelimb cortex. Electrophysiological recordings from the four locations were obtained before lesion and weekly for up to 4 weeks. Our results show that cortical reorganization depends on the type of spinal cord injury, which tends to be more bilateral in full transection while is more unilateral in the model of contusive spinal cord injury. Moreover, in full transection of spinal cord, the deafferented and intact cortex exhibited similar increments of somatosensory evoked responses in both models of spinal cord injury - a feature observed in about 80% of subjects. The other 20% were unaffected by the injury indicating that cortical reorganization does not undergo in all subjects. In addition, we demonstrated an increased probability of triggered up-states in animals with spinal cord injury. This data indicates increased cortical excitability that could be proposed as a new feature of cortical reorganization. Finally, decreased levels of GABA marker GAD67 across cortical layers were only found in those animals with increased somatosensory evoked responses, but not in the unaffected population. In conclusion, cortical reorganization depends on the types of spinal cord injuries, and suggest that the phenomenon is strongly determined by cortical circuits. Moreover, changes in GABAergic transmission at the deprived cortex may be considered one of the mechanisms underlying the process of cortical reorganization and increased excitability.

Astrocytes adjust the dynamic range of cortical network activity to control modality-specific sensory information processing.

Cortical neuron-astrocyte communication in response to peripheral sensory stimulation occurs in a topographic-, frequency-, and intensity-dependent manner. However, the contribution of this functional interaction to the processing of sensory inputs and consequent behavior remains unclear. We investigate the role of astrocytes in sensory information processing at circuit and behavioral levels by monitoring and manipulating astrocytic activity in vivo. We show that astrocytes control the dynamic range of the cortical network activity, optimizing its responsiveness to incoming sensory inputs. The astrocytic modulation of sensory processing contributes to setting the detection threshold for tactile and thermal behavior responses. The mechanism of such astrocytic control is mediated through modulation of inhibitory transmission to adjust the gain and sensitivity of responding networks. These results uncover a role for astrocytes in maintaining the cortical network activity in an optimal range to control behavior associated with specific sensory modalities.

Normal Cortical Myelination in Galectin-4-Deficient Mice.

Myelin, critical for the correct function of the nervous system, is organized in different patterns that can include long non-myelinated axonal segments. How myelin patterning is regulated remains unexplained. The carbohydrate-binding protein galectin-4 (Gal-4) influences oligodendrocyte differentiation in vitro and is associated with non-myelinable axon segments (NMS) in cultured neurons. In consequence, Gal-4 has been proposed as a myelin patterning regulator, although no in vivo studies have corroborated this hypothesis. We used Gal-4-deficient mice (Lgals4-KO) to study the role of Gal-4 in cortical myelination in vivo. We show that cultured neurons of Lgals4-KO mice form NMS that are regulated as in control neurons. In addition, oligodendrocyte/myelin markers expression measured by biochemical and immunochemical means, and cortical myelin microstructure studied by in-depth image analysis appear unaltered in these animals. Consistently, myelin displays an essentially normal function assessed by in vivo electrophysiology and locomotion analyses. In conclusion, cortical myelin of Lgals4-KO mice does not show any significant defect in composition, organization or function, pointing to a negligible role of Gal-4 in myelination in vivo or, as discussed, to unknown mechanisms that compensate its absence.

Cortical layer-specific modulation of neuronal activity after sensory deprivation due to spinal cord injury.

Cortical areas have the capacity of large-scale reorganization following sensory deafferentation. However, it remains unclear whether this phenomenon is a unique process that homogeneously affects the entire deprived cortical region or whether it is susceptible to changes depending on neuronal networks across distinct cortical layers. Here, we studied how the local circuitry within each layer of the deafferented cortex forms the basis for neuroplastic changes after immediate thoracic spinal cord injury (SCI) in anaesthetized rats. In vivo electrophysiological recordings from deafferented hindlimb somatosensory cortex showed that SCI induces layer-specific changes mediating evoked and spontaneous activity. In supragranular layer 2/3, SCI increased gamma oscillations and the ability of these neurons to initiate up-states during spontaneous activity, suggesting an altered corticocortical network and/or intrinsic properties that may serve to maintain the excitability of the cortical column after deafferentation. On the other hand, SCI enhanced the infragranular layers' ability to integrate evoked sensory inputs leading to increased and faster neuronal responses. Delayed evoked response onsets were also observed in layer 5/6, suggesting alterations in thalamocortical connectivity. Altogether, our data indicate that SCI immediately modifies the local circuitry within the deafferented cortex allowing supragranular layers to better integrate spontaneous corticocortical information, thus modifying column excitability, and infragranular layers to better integrate evoked sensory inputs to preserve subcortical outputs. These layer-specific neuronal changes may guide the long-term alterations in neuronal excitability and plasticity associated with the rearrangements of somatosensory networks and the appearance of central sensory pathologies usually associated with spinal cord injury. KEY POINTS: Sensory stimulation of forelimb produces cortical evoked responses in the somatosensory hindlimb cortex in a layer-dependent manner. Spinal cord injury favours the input statistics of corticocortical connections between intact and deafferented cortices. After spinal cord injury supragranular layers exhibit better integration of spontaneous corticocortical information while infragranular layers exhibit better integration of evoked sensory stimulation. Cortical reorganization is a layer-specific phenomenon.

Dual Cortical Plasticity After Spinal Cord Injury.

During cortical development, plasticity reflects the dynamic equilibrium between increasing and decreasing functional connectivity subserved by synaptic sprouting and pruning. After adult cortical deafferentation, plasticity seems to be dominated by increased functional connectivity, leading to the classical expansive reorganization from the intact to the deafferented cortex. In contrast, here we show a striking "decrease" in the fast cortical responses to high-intensity forepaw stimulation 1-3 months after complete thoracic spinal cord transection, as evident in both local field potentials and intracellular in vivo recordings. Importantly, this decrease in fast cortical responses co-exists with an "increase" in cortical activation over slower post-stimulus timescales, as measured by an increased forepaw-to-hindpaw propagation of stimulus-triggered cortical up-states, as well as by the enhanced slow sustained depolarization evoked by high-frequency forepaw stimuli in the deafferented hindpaw cortex. This coincidence of diminished fast cortical responses and enhanced slow cortical activation offers a dual perspective of adult cortical plasticity after spinal cord injury.

Spinal cord injury immediately changes the state of the brain.

Spinal cord injury can produce extensive long-term reorganization of the cerebral cortex. Little is known, however, about the sequence of cortical events starting immediately after the lesion. Here we show that a complete thoracic transection of the spinal cord produces immediate functional reorganization in the primary somatosensory cortex of anesthetized rats. Besides the obvious loss of cortical responses to hindpaw stimuli (below the level of the lesion), cortical responses evoked by forepaw stimuli (above the level of the lesion) markedly increase. Importantly, these increased responses correlate with a slower and overall more silent cortical spontaneous activity, representing a switch to a network state of slow-wave activity similar to that observed during slow-wave sleep. The same immediate cortical changes are observed after reversible pharmacological block of spinal cord conduction, but not after sham. We conclude that the deafferentation due to spinal cord injury can immediately (within minutes) change the state of large cortical networks, and that this state change plays a critical role in the early cortical reorganization after spinal cord injury.