Cortical Networks Relating to Arousal Are Differentially Coupled to Neural Activity and Hemodynamics.

Even in the absence of specific sensory input or a behavioral task, the brain produces structured patterns of activity. This organized activity is modulated by changes in arousal. Here, we use wide-field voltage imaging to establish how arousal relates to cortical network voltage and hemodynamic activity in spontaneously behaving head-fixed male and female mice expressing the voltage-sensitive fluorescent FRET sensor Butterfly 1.2. We find that global voltage and hemodynamic signals are both positively correlated with changes in arousal with a maximum correlation of 0.5 and 0.25, respectively, at a time lag of 0 s. We next show that arousal influences distinct cortical regions for both voltage and hemodynamic signals. These include a broad positive correlation across most sensory-motor cortices extending posteriorly to the primary visual cortex observed in both signals. In contrast, activity in the prefrontal cortex is positively correlated to changes in arousal for the voltage signal while it is a slight net negative correlation observed in the hemodynamic signal. Additionally, we show that coherence between voltage and hemodynamic signals relative to arousal is strongest for slow frequencies below 0.15 Hz and is near zero for frequencies >1 Hz. We finally show that coupling patterns are dependent on the behavioral state of the animal with correlations being driven by periods of increased orofacial movement. Our results indicate that while hemodynamic signals show strong relations to behavior and arousal, these relations are distinct from those observed by voltage activity.

Biomechanical Evaluation of TensionLoc Versus the Double Spike Plate for ACL Graft Tibial Fixation.

Background: The optimal tibial fixation of anterior cruciate ligament (ACL) reconstruction (ACLR) grafts remains controversial. Purpose/Hypothesis: The purpose of this study was to compare the biomechanical characteristics of the TensionLoc (TL) cortical fixation device with the Double Spike Plate (DSP) fixation device for ACL tibial fixation using both bone-patellar tendon-bone (BTB) and quadriceps grafts. It was hypothesized that there would be no differences in biomechanical characteristics between the fixation devices regardless of graft type. Study Design: Controlled laboratory study. Methods: ACLR was performed on 14 matched-pair cadaveric knee specimens-7 pairs using quadriceps grafts (n = 3 male cadaveric knee specimens; n = 4 female cadaveric knee specimens; age, 51 ± 8 years) and 7 pairs using BTB grafts (n = 3 male cadaveric knee specimens; n = 4 female cadaveric knee specimens; age, 50 ± 7 years). One side of each pair was randomized to receive DSP fixation, and the contralateral side received TL fixation. Specimens underwent cyclic ramp loading (10 cycles each at 50-100 N, 50-250 N, and 50-400 N), followed by load-to-failure testing, with the tensile force in line with the tibial tunnel. Results between the 2 fixation types were compared with a paired t test. Results: For the quadriceps graft, there were no significant differences in cyclic loading or load-to-failure characteristics between fixation types (P≥ .092 for all parameters). For the BTB graft, TL fixation resulted in higher stiffness than DSP at all cyclic testing cycles except for cycle 1 during 100-N loading and had lower displacement at 250-N loading (3.4 ± 0.1 vs 5.4 ± 0.3 mm; P = .045). For load to failure, TL fixation resulted in higher stiffness than DSP fixation (232 ± 3.1 vs 188.4 ± 6.4 N/mm; P = .046); however, all other load-to-failure parameters were not statistically different (P≥ .135 for all parameters). Conclusion: With the quadriceps tendon graft, there were no significant differences in biomechanical characteristics between TL and DSP ACL tibial fixations; however, with BTB grafts, the TL tibial fixation demonstrated greater biomechanical integrity than the DSP tibial fixation. Clinical Relevance: The TL fixation device may provide an alternative ACL tibial fixation option for BTB and soft tissue grafts.

Thalamic bursting and the role of timing and synchrony in thalamocortical signaling in the awake mouse.

The thalamus controls transmission of sensory signals from periphery to cortex, ultimately shaping perception. Despite this significant role, dynamic thalamic gating and the consequences for downstream cortical sensory representations have not been well studied in the awake brain. We optogenetically modulated the ventro-posterior-medial thalamus in the vibrissa pathway of the awake mouse and measured spiking activity in the thalamus and activity in primary somatosensory cortex (S1) using extracellular electrophysiology and genetically encoded voltage imaging. Thalamic hyperpolarization significantly enhanced thalamic sensory-evoked bursting; however, surprisingly, the S1 cortical response was not amplified, but instead, timing precision was significantly increased, spatial activation more focused, and there was an increased synchronization of cortical inhibitory neurons. A thalamocortical network model implicates the modulation of precise timing of feedforward thalamic population spiking, presenting a highly sensitive, timing-based gating of sensory signaling to the cortex.

Emerging experience-dependent dynamics in primary somatosensory cortex reflect behavioral adaptation.

Behavioral experience and flexibility are crucial for survival in a constantly changing environment. Despite evolutionary pressures to develop adaptive behavioral strategies in a dynamically changing sensory landscape, the underlying neural correlates have not been well explored. Here, we use genetically encoded voltage imaging to measure signals in primary somatosensory cortex (S1) during sensory learning and behavioral adaptation in the mouse. In response to changing stimulus statistics, mice adopt a strategy that modifies their detection behavior in a context dependent manner as to maintain reward expectation. Surprisingly, neuronal activity in S1 shifts from simply representing stimulus properties to transducing signals necessary for adaptive behavior in an experience dependent manner. Our results suggest that neuronal signals in S1 are part of an adaptive framework that facilitates flexible behavior as individuals gain experience, which could be part of a general scheme that dynamically distributes the neural correlates of behavior during learning.

Rapid Cortical Adaptation and the Role of Thalamic Synchrony during Wakefulness.

Rapid sensory adaptation is observed across all sensory systems, and strongly shapes sensory percepts in complex sensory environments. Yet despite its ubiquity and likely necessity for survival, the mechanistic basis is poorly understood. A wide range of primarily in vitro and anesthetized studies have demonstrated the emergence of adaptation at the level of primary sensory cortex, with only modest signatures in earlier stages of processing. The nature of rapid adaptation and how it shapes sensory representations during wakefulness, and thus the potential role in perceptual adaptation, is underexplored, as are the mechanisms that underlie this phenomenon. To address these knowledge gaps, we recorded spiking activity in primary somatosensory cortex (S1) and the upstream ventral posteromedial (VPm) thalamic nucleus in the vibrissa pathway of awake male and female mice, and quantified responses to whisker stimuli delivered in isolation and embedded in an adapting sensory background. We found that cortical sensory responses were indeed adapted by persistent sensory stimulation; putative excitatory neurons were profoundly adapted, and inhibitory neurons only modestly so. Further optogenetic manipulation experiments and network modeling suggest this largely reflects adaptive changes in synchronous thalamic firing combined with robust engagement of feedforward inhibition, with little contribution from synaptic depression. Taken together, these results suggest that cortical adaptation in the regime explored here results from changes in the timing of thalamic input, and the way in which this differentially impacts cortical excitation and feedforward inhibition, pointing to a prominent role of thalamic gating in rapid adaptation of primary sensory cortex.SIGNIFICANCE STATEMENT Rapid adaptation of sensory activity strongly shapes representations of sensory inputs across all sensory pathways over the timescale of seconds, and has profound effects on sensory perception. Despite its ubiquity and theoretical role in the efficient encoding of complex sensory environments, the mechanistic basis is poorly understood, particularly during wakefulness. In this study in the vibrissa pathway of awake mice, we show that cortical representations of sensory inputs are strongly shaped by rapid adaptation, and that this is mediated primarily by adaptive gating of the thalamic inputs to primary sensory cortex and the differential way in which these inputs engage cortical subpopulations of neurons.

Compensation of physiological motion enables high-yield whole-cell recording in vivo.

BACKGROUND: Whole-cell patch-clamp recording in vivo is the gold-standard method for measuring subthreshold electrophysiology from single cells during behavioural tasks, sensory stimulations, and optogenetic manipulation. However, these recordings require a tight, gigaohm resistance, seal between a glass pipette electrode's aperture and a cell's membrane. These seals are difficult to form, especially in vivo, in part because of a strong dependence on the distance between the pipette aperture and cell membrane. NEW METHOD: We elucidate and utilize this dependency to develop an autonomous method for placement and synchronization of pipette's tip aperture to the membrane of a nearby, moving neuron, which enables high-yield seal formation and subsequent recordings deep in the brain of the living mouse. RESULTS: This synchronization procedure nearly doubles the reported gigaseal yield in the thalamus (>3 mm below the pial surface) from 26 % (n = 17/64) to 48 % (n = 32/66). Whole-cell recording yield improved from 10 % (n = 9/88) to 24 % (n = 18/76) when motion compensation was used during the gigaseal formation. As an example of its application, we utilized this system to investigate the role of the sensory environment and ventral posterior medial region (VPM) projection synchrony on intracellular dynamics in the barrel cortex. COMPARISON WITH EXISTING METHOD(S): Current methods of in vivo whole-cell patch clamping do not synchronize the position of the pipette to motion of the cell. CONCLUSIONS: This method results in substantially greater subcortical whole-cell recording yield than previously reported and thus makes pan-brain whole-cell electrophysiology practical in the living mouse brain.

Unilateral Optogenetic Inhibition and Excitation of Basal Ganglia Output Affect Directional Lick Choices and Movement Initiation in Mice.

Models of basal ganglia (BG) function predict that tonic inhibitory output to motor thalamus (MT) suppresses unwanted movements, and that a decrease in such activity leads to action selection. Further, for unilateral activity changes in the BG, a lateralized effect on contralateral movements can be expected due to ipsilateral thalamocortical connectivity. However, a direct test of these outcomes of thalamic inhibition has not been performed. To conduct such a direct test, we utilized rapid optogenetic activation and inactivation of the GABAergic output of the substantia nigra pars reticulata (SNr) to MT in male and female mice that were trained in a sensory cued left/right licking task. Directional licking tasks have previously been shown to depend on a thalamocortical feedback loop between ventromedial MT and antero-lateral premotor cortex. In confirmation of model predictions, we found that unilateral optogenetic inhibition of GABAergic output from the SNr, during ipsilaterally cued trials, biased decision making towards a contralateral lick without affecting motor performance. In contrast, optogenetic excitation of SNr terminals in MT resulted in an opposite bias towards the ipsilateral direction confirming a bidirectional effect of tonic nigral output on directional decision making. However, direct optogenetic excitation of neurons in the SNr resulted in bilateral movement suppression, which is in agreement with previous results that show such suppression for nigral terminals in the superior colliculus (SC), which receives a bilateral projection from SNr.

Stimulus Context and Reward Contingency Induce Behavioral Adaptation in a Rodent Tactile Detection Task.

Behavioral adaptation is a prerequisite for survival in a constantly changing sensory environment, but the underlying strategies and relevant variables driving adaptive behavior are not well understood. Many learning models and neural theories consider probabilistic computations as an efficient way to solve a variety of tasks, especially if uncertainty is involved. Although this suggests a possible role for probabilistic inference and expectation in adaptive behaviors, there is little if any evidence of this relationship experimentally. Here, we investigated adaptive behavior in the rat model by using a well controlled behavioral paradigm within a psychophysical framework to predict and quantify changes in performance of animals trained on a simple whisker-based detection task. The sensory environment of the task was changed by transforming the probabilistic distribution of whisker deflection amplitudes systematically while measuring the animal's detection performance and corresponding rate of accumulated reward. We show that the psychometric function deviates significantly and reversibly depending on the probabilistic distribution of stimuli. This change in performance relates to accumulating a constant reward count across trials, yet it is exempt from changes in reward volume. Our simple model of reward accumulation captures the observed change in psychometric sensitivity and predicts a strategy seeking to maintain reward expectation across trials in the face of the changing stimulus distribution. We conclude that rats are able maintain a constant payoff under changing sensory conditions by flexibly adjusting their behavioral strategy. Our findings suggest the existence of an internal probabilistic model that facilitates behavioral adaptation when sensory demands change.SIGNIFICANCE STATEMENT The strategy animals use to deal with a complex and ever-changing world is a key to understanding natural behavior. This study provides evidence that rodent behavioral performance is highly flexible in the face of a changing stimulus distribution, consistent with a strategy to maintain a desired accumulation of reward.

Erratum: Genetically expressed voltage sensor ArcLight for imaging large scale cortical activity in the anesthetized and awake mouse (erratum).

[This corrects the article DOI: 10.1117/1.NPh.4.3.031212.].

Genetically expressed voltage sensor ArcLight for imaging large scale cortical activity in the anesthetized and awake mouse.

With the recent breakthrough in genetically expressed voltage indicators (GEVIs), there has been a tremendous demand to determine the capabilities of these sensors in vivo. Novel voltage sensitive fluorescent proteins allow for direct measurement of neuron membrane potential changes through changes in fluorescence. Here, we utilized ArcLight, a recently developed GEVI, and examined the functional characteristics in the widely used mouse somatosensory whisker pathway. We measured the resulting evoked fluorescence using a wide-field microscope and a CCD camera at 200 Hz, which enabled voltage recordings over the entire cortical region with high temporal resolution. We found that ArcLight produced a fluorescent response in the S1 barrel cortex during sensory stimulation at single whisker resolution. During wide-field cortical imaging, we encountered substantial hemodynamic noise that required additional post hoc processing through noise subtraction techniques. Over a period of 28 days, we found clear and consistent ArcLight fluorescence responses to a simple sensory input. Finally, we demonstrated the use of ArcLight to resolve cortical S1 sensory responses in the awake mouse. Taken together, our results demonstrate the feasibility of ArcLight as a measurement tool for mesoscopic, chronic imaging.

Biomechanical comparison of the FasT-Fix meniscal repair suture system with vertical mattress sutures and meniscus arrows.

BACKGROUND: A meniscal repair technique that combines the strength of vertical mattress sutures and the decreased tissue morbidity of an all-inside technique would be advantageous. HYPOTHESIS: The FasT-Fix Meniscal Repair Suture System will provide load at failure, stiffness, and displacement equivalent to that of vertical mattress sutures and superior to that of Meniscus Arrows. STUDY DESIGN: In vitro biomechanical study. METHODS: After repair of a 2-cm vertical longitudinal medial meniscal lesion, three groups of six human cadaveric knees were biomechanically tested in a random order on a servohydraulic device, and three groups of five specimens underwent cyclic loading. RESULTS: Specimens repaired with Meniscus Arrows had reduced load at failure, stiffness, and displacement, but there were no differences between the FasT-Fix and vertical mattress suture methods. During cyclic loading, specimens repaired with two Meniscus Arrows failed before test completion, whereas specimens repaired with two vertical mattress sutures (6.0 +/- 3.7 mm) or with two FasT-Fix implants (5.1 +/- 1.4 mm) maintained fixation with comparable displacements. CONCLUSIONS: The FasT-Fix provided load at failure, stiffness, and displacement comparable with that of vertical mattress sutures. CLINICAL RELEVANCE: The results suggest that the FasT-Fix may be preferable to Meniscus Arrows for meniscal repair with minimal associated tissue morbidity.

Complications of tibial osteotomies in children with comorbidities.

Tibial osteotomies in children have been associated with a number of complications. A retrospective review of 116 children who had 129 tibial osteotomies was performed to assess these complications at our institution. Results showed that there were 35 cases of wound problems, 6 cases of recurrence/reoperation, 5 cases of delayed union, 2 cases with transient peroneal nerve palsy, 1 case of nonunion, and 1 case of mal-union. Patients having certain comorbidities had a higher frequency of complications. There were no significant differences between the location of the tibial osteotomy (proximal or distal) and the incidence of complication. External fixation was associated with a lower incidence of complications than the use of pins and casting. Although our results demonstrate an overall low complication rate, there is a significant association between complications and comorbid conditions. This highlights the need to recognize comorbidities preoperatively and the potential of increased postoperative complications.

Medial collateral ligament reconstruction with allograft using a double-bundle technique.

Medial collateral ligament (MCL) reconstruction has been a topic of controversy in regard to the need for surgical reconstruction as well as the type of surgical reconstruction to be performed. Combined anterior cruciate ligament (ACL) and MCL reconstruction has been found to be associated with a higher incidence of postoperative arthrofibrosis than isolated ACL reconstruction; performing these reconstructions in a staged format has been proposed to avoid this devastating complication. We present a technique for MCL reconstruction that physiometrically re-establishes both anterior and posterior stabilizing components of the MCL and is performed with a limited soft-tissue dissection. This technique can easily be combined with an ACL allograft or hamstring reconstruction without need for staged or significantly delayed procedure. The technical details of this technique allow for stable fixation of an allograft reconstruction to allow for immediate postoperative knee range of motion with low patient morbidity because of the limited surgical approach.