Locomotion activates PKA through dopamine and adenosine in striatal neurons.

The canonical model of striatal function predicts that animal locomotion is associated with the opposing regulation of protein kinase A (PKA) in direct and indirect pathway striatal spiny projection neurons (SPNs) by dopamine1-7. However, the precise dynamics of PKA in dorsolateral SPNs during locomotion remain to be determined. It is also unclear whether other neuromodulators are involved. Here we show that PKA activity in both types of SPNs is essential for normal locomotion. Using two-photon fluorescence lifetime imaging8-10 of a PKA sensor10 through gradient index lenses, we measured PKA activity within individual SPNs of the mouse dorsolateral striatum during locomotion. Consistent with the canonical view, dopamine activated PKA activity in direct pathway SPNs during locomotion through the dopamine D1 receptor. However, indirect pathway SPNs exhibited a greater increase in PKA activity, which was largely abolished through the blockade of adenosine A2A receptors. In agreement with these results, fibre photometry measurements of an adenosine sensor11 revealed an acute increase in extracellular adenosine during locomotion. Functionally, antagonism of dopamine or adenosine receptors resulted in distinct changes in SPN PKA activity, neuronal activity and locomotion. Together, our results suggest that acute adenosine accumulation interplays with dopamine release to orchestrate PKA activity in SPNs and proper striatal function during animal locomotion.

Sensitive genetically encoded sensors for population and subcellular imaging of cAMP in vivo.

Cyclic adenosine monophosphate (cAMP) signaling integrates information from diverse G-protein-coupled receptors, such as neuromodulator receptors, to regulate pivotal biological processes in a cellular-specific and subcellular-specific manner. However, in vivo cellular-resolution imaging of cAMP dynamics remains challenging. Here, we screen existing genetically encoded cAMP sensors and further develop the best performer to derive three improved variants, called cAMPFIREs. Compared with their parental sensor, these sensors exhibit up to 10-fold increased sensitivity to cAMP and a cytosolic distribution. cAMPFIREs are compatible with both ratiometric and fluorescence lifetime imaging and can detect cAMP dynamics elicited by norepinephrine at physiologically relevant, nanomolar concentrations. Imaging of cAMPFIREs in awake mice reveals tonic levels of cAMP in cortical neurons that are associated with wakefulness, modulated by opioids, and differentially regulated across subcellular compartments. Furthermore, enforced locomotion elicits neuron-specific, bidirectional cAMP dynamics. cAMPFIREs also function in Drosophila. Overall, cAMPFIREs may have broad applicability for studying intracellular signaling in vivo.

Genetically encoded fluorescent sensors for imaging neuronal dynamics in vivo.

The brain relies on many forms of dynamic activities in individual neurons, from synaptic transmission to electrical activity and intracellular signaling events. Monitoring these neuronal activities with high spatiotemporal resolution in the context of animal behavior is a necessary step to achieve a mechanistic understanding of brain function. With the rapid development and dissemination of highly optimized genetically encoded fluorescent sensors, a growing number of brain activities can now be visualized in vivo. To date, cellular calcium imaging, which has been largely used as a proxy for electrical activity, has become a mainstay in systems neuroscience. While challenges remain, voltage imaging of neural populations is now possible. In addition, it is becoming increasingly practical to image over half a dozen neurotransmitters, as well as certain intracellular signaling and metabolic activities. These new capabilities enable neuroscientists to test previously unattainable hypotheses and questions. This review summarizes recent progress in the development and delivery of genetically encoded fluorescent sensors, and highlights example applications in the context of in vivo imaging.

Photon-efficient 3D reconstruction employing a edge enhancement method.

Photon-efficient 3D reconstruction under sparse photon conditions remains challenges. Especially for scene edge locations, the light scattering results in a weaker echo signal than non-edge locations. Depth images can be viewed as smooth regions stitched together by edge segmentation, yet none of the existing methods focus on how to improve the accuracy of edge reconstruction when performing 3D reconstruction. Moreover, the impact of edge reconstruction to overall depth reconstruction hasn't been investigated. In this paper, we explore how to improve the edge reconstruction accuracy from various aspects such as improving the network structure, employing hybrid loss functions and taking advantages of the non-local correlation of SPAD measurements. Meanwhile, we investigate the correlation between the edge reconstruction accuracy and the reconstruction accuracy of overall depth based on quantitative metrics. The experimental results show that the proposed method achieves superior performance in both edge reconstruction and overall depth reconstruction compared with other state-of-the-art methods. Besides, it proves that the improvement of edge reconstruction accuracy promotes the reconstruction accuracy of depth map.

Static coded aperture in robotic X-ray tomography systems.

Coded aperture X-ray computed tomography is a computational imaging technique capable of reconstructing inner structures of an object from a reduced set of X-ray projection measurements. Coded apertures are placed in front of the X-ray sources from different views and thus significantly reduce the radiation dose. This paper introduces coded aperture X-ray computed tomography for robotic X-ray systems which offer positioning flexibility. While single coded-aperture 3D tomography was recently introduced for standard trajectory CT scanning, it is shown that significant gains in imaging performance can be attained by simple modifications in the CT scanning trajectories enabled by emerging dual robotic CT systems. In particular, the subject is fixed on a plane and the CT system uniformly rotates around the r -axis which is misaligned with the coordinate axes. A single stationary coded aperture is placed on front of the robotic X-ray source above the plane and the corresponding X-ray projections are measured by a two-dimensional detector on the second arm of the robotic system. The compressive measurements with misalignment enable the reconstruction of high-resolution three-dimensional volumetric images from the low-resolution coded projections on the detector at a sub-sampling rate. An efficient algorithm is proposed to generate the rotation matrix with two basic sub-matrices and thus the forward model is formulated. The stationary coded aperture is designed based on the Pearson product-moment correlation coefficient analysis and the direct binary search algorithm is used to obtain the optimized coded aperture. Simulations using simulated datasets show significant gains in reconstruction performance compared to conventional coded aperture CT systems.

Frequency upconversion imaging based on Hadamard coding.

A frequency upconversion imaging based on Hadamard coding is presented to remove the distorting effect on condition that the pump beam is tightly focused to optimize the conversion efficiency. The distortion caused by the convolution between the object field and the pump field is ascribed to the point spread function effect. In order to remove the blurring in an upconversion imaging system optimized by tight focused pump, the object is encoded by measurement matrices and the corresponding intensity of the converted field is measured. Thus the intensity distribution of the object can be calculated accurately by the measurements and the measurement matrix. The signal-to-noise ratio (SNR) is improved by employing the Hadamard matrix since the intensity of measured converted signal is far larger than the intensity of each pixel. The experimental results show the proposed method removes the distorting effect caused by the convolution. The converted image still has sharp edges on condition that the conversion efficiency is optimized by tight focusing the pump beam.

Single-snapshot X-ray imaging for nonlinear compressive tomosynthesis.

Traditional compressive X-ray tomosynthesis uses sequential illumination to interrogate the object, leading to long scanning time and image distortion due to the object variation. This paper proposes a single-snapshot compressive tomosynthesis imaging approach, where the object is simultaneously illuminated by multiple X-ray emitters equipped with coded apertures. Based on rank, intensity and sparsity prior models, a nonlinear image reconstruction framework is established. The coded aperture patterns are optimized based on uniform sensing criteria. Then, a modified split Bregman algorithm is developed to reconstruct the object from the set of nonlinear compressive measurements. It is shown that the proposed method can be used to reduce the inspection time and achieve robust reconstruction with respect to shape variation or motion of objects.

Source and coded aperture joint optimization for compressive X-ray tomosynthesis.

Compressive X-ray tomosynthesis is an emerging technique to reconstruct three-dimensional (3D) objects from two-dimensional projection measurements generated by a set of spatially distributed X-ray sources, where coded apertures are used in front of each source to modulate a set of X-rays to interrogate an object with a reduced radiation dose without loss of image reconstruction quality. The reconstruction performance in compressive tomosynthesis is influenced by several factors including the locations of the X-ray sources, their incident angles, and the coded apertures that determine the structured illumination patterns. This paper develops a source and coded aperture joint optimization (SCO) approach to improve the image reconstruction performance of compressive X-ray tomosynthesis. Based on compressive sensing theory, the synergy among the source pattern, source orientation, and the coded apertures is utilized to minimize the coherence of the sensing matrix of the imaging system. In concert with a gradient-based optimization algorithm, regularization methods are used to reduce the convergence error and achieve uniform sensing of the object under inspection. Compared to the optimization of either the source orientation, or the coded aperture individually, the proposed method effectively increases the degree of optimization freedom, and thus achieves considerable improvement in the 3D imaging reconstruction accuracy.

Fast optimization of coded apertures in X-ray computed tomography.

Coded aperture X-ray computed tomography (CAXCT) is a novel X-ray imaging system capable of reconstructing high quality images from a reduced set of measurements. Coded apertures are placed in front of the X-ray source in CAXCT so as to obtain patterned projections onto a detector array. Then, compressive sensing (CS) reconstruction algorithms are used to reconstruct the linear attenuation coefficients. The coded aperture is an important factor that influences the point spread function (PSF), which in turn determines the capability to sample the linear attenuation coefficients of the object. A coded aperture optimization approach was recently proposed based on the coherence of the system matrix; however, this algorithm is memory intensive and it is not able to optimize the coded apertures for large image sizes required in many applications. This paper introduces a significantly more efficient approach for coded aperture optimization that reduces the memory requirements and the execution time by orders of magnitude. The features are defined as the inner product of the vectors representing the geometric paths of the X-rays with the sparse basis representation of the object; therefore, the algorithm aims to find a subset of features that minimizes the information loss compared to the complete set of projections. This subset corresponds to the unblocking elements in the optimized coded apertures. The proposed approach solves the memory and runtime limitations of the previously proposed algorithm and provides a significant gain in the reconstruction image quality compared to that attained by random coded apertures in both simulated datasets and real datasets.

Target-switched triplex nanotweezer and synergic fluorophore translocation for highly selective melamine assay.

This paper describes a triplex DNA nanotweezer to specifically capture melamine (MEL). The triplex-forming oligonucleotide (TFO) arm can be switched from the open state to the closed state once MEL binds to the abasic site (AP site) in duplex via the bifacial hydrogen bonding with thymines. Following this nanotweezer operation, the AP site-bound fluorophore is translocated to the terminal triplet to subsequently light up the nanotweezer. The TFO arm is found to be pivotal for permitting the AP site binding. The synergic processes of target competition and fluorophore translocation support a high selectivity for the MEL assay even against the inherent adenosine and the MEL hydrolysis products. Chelerythrine is employed as the fluorescent probe. The detection limit of MEL was estimated to be about 140 nM assuming a signal-to-noise ratio of 3. It was applied to the determination of MEL in spiked milk samples without any separation procedure. Conceivably, this method opens a new avenue towards highly selective triplex-based sensors by making use of other commercially available DNA modifications for recognizing other analytes. Graphical abstract Schematic presentation of a triplex nanotweezer with an open-to-close conversion upon the abasic site binding of melamine. The assay is based on a synergic fluorophore translocation. The corresponding duplex otherwise shows no binding with melamine. Chelerythrine (CHE) with a yellow-green emission peaking at 544 nm is employed as the fluorescent probe.

Analysis of the main constituents of Changshu tablet and its spasmolysis effect against contraction induced by acetylcholine in the rat-isolated intestinal smooth muscle.

The Changshu tablet (CST), one kind of Chinese patent medicine with astringent to the intestine and relieving diarrhea, was made by the root of Rose odorata Sweet var. gigantean (Coll.et Hemsl.) Rehd.et Wils. Although CST has a long history of clinical application, but the research of its chemical composition is less. So the objective of this study was to investigate the main constituents and preliminarily research its effect of the contraction of isolated intestine in vitro. The contents of total polyphenols (126.23mg/g) and total triterpenoids (132.75mg/g) in CST were determined by ultraviolet spectrophotometry. Procyanidin B3, epigallo catechin, catechin, epicatechin, (-)-fisetinidol-(4α, 8)-(-)-catechin, (4α, 8)-(-)-fisetinidol-(-)-epicatechins and (+)-guibourtinidol-(4ß, 8)-epicatechin were identified and determined by high performance liquid chromatography and their contents were distributed from 0.04mg/g to 1.46 mg/g. CST showed significant inhibitory effect against acetylcholine-induced contraction on the rat-isolated intestinal smooth muscle with a dose-dependent manner from 0.06 to 0.6mg/mL. The maxim inhibition rates of CST on duodenum, jejunum, ileum and colon were 65.70±3.47%, 79.74±1.27%, 58.90±1.87% and 45.75±2.21% respectively. These results indicated that CST has a spasmolytic role in gastrointestinal motility which was probably mediated through inhibition of muscarinic receptors. All these findings promote the improvement of the quality control standard of CST and provide pharmacological foundation for clinical application of CST in gastrointestinal tract.

Optogenetic Control of Mouse Outer Hair Cells.

Normal hearing in mammals depends on sound amplification by outer hair cells (OHCs) presumably by their somatic motility and force production. However, the role of OHC force production in cochlear amplification and frequency tuning are not yet fully understood. Currently, available OHC manipulation techniques for physiological or clinical studies are limited by their invasive nature, lack of precision, and poor temporal-spatial resolution. To overcome these limitations, we explored an optogenetic approach based on channelrhodopsin 2 (ChR-2), a direct light-activated nonselective cation channel originally discovered in Chlamydomonas reinhardtii. Three approaches were compared: 1) adeno-associated virus-mediated in utero transfer of the ChR-2 gene into the developing murine otocyst, 2) expression of ChR-2(H134R) in an auditory cell line (HEI-OC1), and 3) expression of ChR-2 in the OHCs of a mouse line carrying a ChR-2 conditional allele. Whole cell recording showed that blue light (470 nm) elicited the typical nonselective cation current of ChR-2 with reversal potential around zero in both mouse OHCs and HEI-OC1 cells and generated depolarization in both cell types. In addition, pulsed light stimulation (10 Hz) elicited a 1:1 repetitive depolarization and ChR-2 currents in mouse OHCs and HEI-OC1 cells, respectively. The time constant of depolarization in OHCs, 1.45 ms, is 10 times faster than HEI-OC1 cells, which allowed light stimulation up to rates of 10/s to elicit corresponding membrane potential changes. Our study demonstrates that ChR-2 can successfully be expressed in mouse OHCs and HEI-OC1 cells and that these present a typical light-sensitive current and depolarization. However, the amount of ChR-2 current induced in our in vivo experiments was insufficient to result in measurable cochlear effects.

Adaptive compressed photon counting 3D imaging based on wavelet trees and depth map sparse representation.

We demonstrate a photon counting 3D imaging system with short-pulsed structured illumination and a single-pixel photon counting detector. The proposed multiresolution photon counting 3D imaging technique acquires a high-resolution 3D image from a coarse image and details at successfully finer resolution sampled along the wavelet trees and their depth map sparse representations. Both the required measurements and the reconstruction time can be significant reduced, which makes the proposed technique suitable for scenes with high spatial resolution. The experimental results indicate that both the reflectivity and depth map of a scene at resolutions up to 512×512 pixels can be acquired and retrieved with practical times as low as 17.5 seconds. In addition, we demonstrate that this technique has ability to image in presence of partially-transmissive occluders, and to directly acquire novelty images to find changes in a scene.

Live imaging of endogenous PSD-95 using ENABLED: a conditional strategy to fluorescently label endogenous proteins.

Stoichiometric labeling of endogenous synaptic proteins for high-contrast live-cell imaging in brain tissue remains challenging. Here, we describe a conditional mouse genetic strategy termed endogenous labeling via exon duplication (ENABLED), which can be used to fluorescently label endogenous proteins with near ideal properties in all neurons, a sparse subset of neurons, or specific neuronal subtypes. We used this method to label the postsynaptic density protein PSD-95 with mVenus without overexpression side effects. We demonstrated that mVenus-tagged PSD-95 is functionally equivalent to wild-type PSD-95 and that PSD-95 is present in nearly all dendritic spines in CA1 neurons. Within spines, while PSD-95 exhibited low mobility under basal conditions, its levels could be regulated by chronic changes in neuronal activity. Notably, labeled PSD-95 also allowed us to visualize and unambiguously examine otherwise-unidentifiable excitatory shaft synapses in aspiny neurons, such as parvalbumin-positive interneurons and dopaminergic neurons. Our results demonstrate that the ENABLED strategy provides a valuable new approach to study the dynamics of endogenous synaptic proteins in vivo.

The subcellular organization of neocortical excitatory connections.

Understanding cortical circuits will require mapping the connections between specific populations of neurons, as well as determining the dendritic locations where the synapses occur. The dendrites of individual cortical neurons overlap with numerous types of local and long-range excitatory axons, but axodendritic overlap is not always a good predictor of actual connection strength. Here we developed an efficient channelrhodopsin-2 (ChR2)-assisted method to map the spatial distribution of synaptic inputs, defined by presynaptic ChR2 expression, within the dendritic arborizations of recorded neurons. We expressed ChR2 in two thalamic nuclei, the whisker motor cortex and local excitatory neurons and mapped their synapses with pyramidal neurons in layers 3, 5A and 5B (L3, L5A and L5B) in the mouse barrel cortex. Within the dendritic arborizations of L3 cells, individual inputs impinged onto distinct single domains. These domains were arrayed in an orderly, monotonic pattern along the apical axis: axons from more central origins targeted progressively higher regions of the apical dendrites. In L5 arborizations, different inputs targeted separate basal and apical domains. Input to L3 and L5 dendrites in L1 was related to whisker movement and position, suggesting that these signals have a role in controlling the gain of their target neurons. Our experiments reveal high specificity in the subcellular organization of excitatory circuits.

Organization of cortical and thalamic input to pyramidal neurons in mouse motor cortex.

Determining how long-range synaptic inputs engage pyramidal neurons in primary motor cortex (M1) is important for understanding circuit mechanisms involved in regulating movement. We used channelrhodopsin-2-assisted circuit mapping to characterize the long-range excitatory synaptic connections made by multiple cortical and thalamic areas onto pyramidal neurons in mouse vibrissal motor cortex (vM1). Each projection innervated vM1 pyramidal neurons with a unique laminar profile. Collectively, the profiles for different sources of input partially overlapped and spanned all cortical layers. Specifically, orbital cortex (OC) inputs primarily targeted neurons in L6. Secondary motor cortex (M2) inputs excited neurons mainly in L5B, including pyramidal tract neurons. In contrast, thalamocortical inputs from anterior motor-related thalamic regions, including VA/VL (ventral anterior thalamic nucleus/ventrolateral thalamic nucleus), targeted neurons in L2/3 through L5B, but avoided L6. Inputs from posterior sensory-related thalamic areas, including POm (posterior thalamic nuclear group), targeted neurons only in the upper layers (L2/3 and L5A), similar to inputs from somatosensory (barrel) cortex. Our results show that long-range excitatory inputs target vM1 pyramidal neurons in a layer-specific manner. Inputs from sensory-related cortical and thalamic areas preferentially target the upper-layer pyramidal neurons in vM1. In contrast, inputs from OC and M2, areas associated with volitional and cognitive aspects of movements, bypass local circuitry and have direct monosynaptic access to neurons projecting to brainstem and thalamus.

A role for myosin VI in the localization of axonal proteins.

In neurons polarized trafficking of vesicle-bound membrane proteins gives rise to the distinct molecular composition and functional properties of axons and dendrites. Despite their central role in shaping neuronal form and function, surprisingly little is known about the molecular processes that mediate polarized targeting of neuronal proteins. Recently, the plus-end-directed motor Myosin Va was shown to play a critical role in targeting of transmembrane proteins to dendrites; however, the role of myosin motors in axonal targeting is unknown. Here we show that Myosin VI, a minus-end-directed motor, plays a vital role in the enrichment of proteins on the surface of axons. Engineering non-neuronal proteins to interact with Myosin VI causes them to become highly concentrated at the axonal surface in dissociated rat cortical neurons. Furthermore, disruption of either Myosin VI function or expression leads to aberrant dendritic localization of axonal proteins. Myosin VI mediates the enrichment of proteins on the axonal surface at least in part by stimulating dendrite-specific endocytosis, a mechanism that has been shown to underlie the localization of many axonal proteins. In addition, a version of Channelrhodopsin 2 that was engineered to bind to Myosin VI is concentrated at the surface of the axon of cortical neurons in mice in vivo, suggesting that it could be a useful tool for probing circuit structure and function. Together, our results indicate that myosins help shape the polarized distributions of both axonal and dendritic proteins.

Fine-tuning TrailMap: The utility of transfer learning to improve the performance of deep learning in axon segmentation of light-sheet microscopy images.

Light-sheet microscopy has made possible the 3D imaging of both fixed and live biological tissue, with samples as large as the entire mouse brain. However, segmentation and quantification of that data remains a time-consuming manual undertaking. Machine learning methods promise the possibility of automating this process. This study seeks to advance the performance of prior models through optimizing transfer learning. We fine-tuned the existing TrailMap model using expert-labeled data from noradrenergic axonal structures in the mouse brain. By changing the cross-entropy weights and using augmentation, we demonstrate a generally improved adjusted F1-score over using the originally trained TrailMap model within our test datasets.

Valence-arousal classification of emotion evoked by Chinese ancient-style music using 1D-CNN-BiLSTM model on EEG signals for college students.

During the COVID-19 pandemic, young people are using multimedia content more frequently to communicate with each other on Internet platforms. Among them, music, as psychological support for a lonely life in this special period, is a powerful tool for emotional self-regulation and getting rid of loneliness. More and more attention has been paid to the music recommender system based on emotion. In recent years, Chinese music has tended to be considered an independent genre. Chinese ancient-style music is one of the new folk music styles in Chinese music and is becoming more and more popular among young people. The complexity of Chinese-style music brings significant challenges to the quantitative calculation of music. To effectively solve the problem of emotion classification in music information search, emotion is often characterized by valence and arousal. This paper focuses on the valence and arousal classification of Chinese ancient-style music-evoked emotion. It proposes a hybrid one-dimensional convolutional neural network and bidirectional and unidirectional long short-term memory model (1D-CNN-BiLSTM). And a self-acquisition EEG dataset for Chinese college students was designed to classify music-induced emotion by valence-arousal based on EEG. In addition to that, the proposed 1D-CNN-BILSTM model verified the performance of public datasets DEAP and DREAMER, as well as the self-acquisition dataset DESC. The experimental results show that, compared with traditional LSTM and 1D-CNN-LSTM models, the proposed method has the highest accuracy in the valence classification task of music-induced emotion, reaching 94.85%, 98.41%, and 99.27%, respectively. The accuracy of the arousal classification task also gained 93.40%, 98.23%, and 99.20%, respectively. In addition, compared with the positive valence classification results of emotion, this method has obvious advantages in negative valence classification. This study provides a computational classification model for a music recommender system with emotion. It also provides some theoretical support for the brain-computer interactive (BCI) application products of Chinese ancient-style music which is popular among young people.

Treatment of paradoxical eczematous eruption in psoriasis treated with secukinumab: A case report.

RATIONALE: Eczematous eruption is an increasingly recognized form of drug-related eruption, typically reported in association with interleukin 17 (IL-17)A inhibitors. However, severe paradoxical eczematous eruption due to IL-17A inhibitors has been rarely reported. Herein, we reported a case of a man with severe psoriasis with erythematous scaly plaques on the scalp, trunk, and arms and legs after the administration of secukinumab was initiated. PATIENT CONCERNS: We reported a case of a 20-year-old man with severe psoriasis with erythematous scaly plaques on the scalp, trunk, and arms and legs after the administration of secukinumab was initiated. A skin biopsy was performed. It revealed spongiotic dermatitis consistent with eczematous reaction. Direct and indirect immunofluorescence assays were negative. DIAGNOSES: He was diagnosed with eczematous eruption. INTERVENTIONS: Discontinuation of secukinumab and administration of cyclosporine and prednisone were considered. OUTCOMES: Significant improvement was observed, with no adverse events. CONCLUSION: Our case shows that eczematous eruption can paradoxically occur in patients on IL-17A inhibitors and this report is expected to increase awareness of the rising number of cutaneous eruptions related to biological agents.