Tools for interfacing, extracting, and analyzing neural signals using wide-field fluorescence imaging and optogenetics in awake behaving mice

Imaging of multiple cells has rapidly multiplied the rate of data acquisition as well as our knowledge of the complex dynamics within the mammalian brain. The process of data acquisition has been dramatically enhanced with highly affordable, sensitive image sensors enable high-throughput detection of neural activity in intact animals. Genetically encoded calcium sensors deliver a substantial boost in signal strength and in combination with equally critical advances in the size, speed, and sensitivity of image sensors available in scientific cameras enables high-throughput detection of neural activity in behaving animals using traditional wide-field fluorescence microscopy. However, the tremendous increase in data flow presents challenges to processing, analysis, and storage of captured video, and prompts a reexamination of traditional routines used to process data in neuroscience and now demand improvements in both our hardware and software applications for processing, analyzing, and storing captured video. This project demonstrates the ease with which a dependable and affordable wide-field fluorescence imaging system can be assembled and integrated with behavior control and monitoring system such as found in a typical neuroscience laboratory. An Open-source MATLAB toolbox is employed to efficiently analyze and visualize large imaging data sets in a manner that is both interactive and fully automated. This software package provides a library of image pre-processing routines optimized for batch-processing of continuous functional fluorescence video, and additionally automates a fast unsupervised ROI detection and signal extraction routine. Further, an extension of this toolbox that uses GPU programming to process streaming video, enabling the identification, segmentation and extraction of neural activity signals on-line is described in which specific algorithms improve signal specificity and image quality at the single cell level in a behaving animal. This project describes the strategic ingredients for transforming a large bulk flow of raw continuous video into proportionally informative images and knowledge

Striatal cholinergic interneurons generate beta and gamma oscillations in the corticostriatal circuit and produce motor deficits

Cortico-basal ganglia-thalamic (CBT) neural circuits are critical modulators of cognitive and motor function. When compromised, these circuits contribute to neurological and psychiatric disorders, such as Parkinson's disease (PD). In PD, motor deficits correlate with the emergence of exaggerated beta frequency (15-30 Hz) oscillations throughout the CBT network. However, little is known about how specific cell types within individual CBT brain regions support the generation, propagation, and interaction of oscillatory dynamics throughout the CBT circuit or how specific oscillatory dynamics are related to motor function. Here, we investigated the role of striatal cholinergic interneurons (SChIs) in generating beta and gamma oscillations in cortical-striatal circuits and in influencing movement behavior. We found that selective stimulation of SChIs via optogenetics in normal mice robustly and reversibly amplified beta and gamma oscillations that are supported by distinct mechanisms within striatal-cortical circuits. Whereas beta oscillations are supported robustly in the striatum and all layers of primary motor cortex (M1) through a muscarinic-receptor mediated mechanism, gamma oscillations are largely restricted to the striatum and the deeper layers of M1. Finally, SChI activation led to parkinsonian-like motor deficits in otherwise normal mice. These results highlight the important role of striatal cholinergic interneurons in supporting oscillations in the CBT network that are closely related to movement and parkinsonian motor symptoms.DP2 NS082126 - NINDS NIH HHS; R01 NS081716 - NINDS NIH HHS; R21 NS078660 - NINDS NIH HHShttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4896681/Published versio

Unique contributions of parvalbumin and cholinergic interneurons in organizing striatal networks during movement

Published in final edited form as: Nat Neurosci. 2019 April ; 22(4): 586–597. doi:10.1038/s41593-019-0341-3.Striatal pavalbumin (PV) and cholinergic (CHI) interneurons are poised to play major roles in behavior by coordinating the networks of medium spiny cells that relay motor output. However, the small numbers and scattered distribution of these cells has made it difficult to directly assess their contribution to activity in networks of MSNs during behavior. Here, we build upon recent improvements in single cell calcium imaging combined with optogenetics to test the capacity of PVs and CHIs to affect MSN activity and behavior in mice engaged in voluntarily locomotion. We find that PVs and CHIs have unique effects on MSN activity and dissociable roles in supporting movement. PV cells facilitate movement by refining the activation of MSN networks responsible for movement execution. CHIs, in contrast, synchronize activity within MSN networks to signal the end of a movement bout. These results provide new insights into the striatal network activity that supports movement.Accepted manuscrip

Self-maintaining or continuously refreshed? The genetic structure of Euphausia lucens populations in the Benguela upwelling ecosystem

Populations of Euphausia lucens over the shelf of the southern Benguela upwelling region could be self-maintaining. Alternatively, they could be continually refreshed by expatriates from the SW Atlantic that enter the system via South Atlantic Central Water in the south, before developing and then being lost through advection off Namibia. These two hypotheses are investigated here by examining geographic heterogeneity and molecular variation (cox1 and ND1) of the species across its distributional range in the Southern Hemisphere. Comparisons are made with E. vallentini, which is assumed to show panmixia associated with its circumglobal distribution between 50 and 60°S. Phylogenetic analysis with mitochondrial 16S ribosomal RNA and cytochrome oxidase 1 (cox1) confirmed that E. lucens and E. vallentinirepresent sister taxa. Strong geographic structuring of cox1 and ND1 mtDNA genetic variation by ocean basin was recorded in E. lucens, indicating that neritic populations off South Africa are likely self-maintaining. This contrasts with the results for E. vallentini, which appears to occur as a single panmictic population across its distributional range. These differences are likely related to the habitats (neritic, E. lucens; oceanic, E. vallentini) occupied by each species. The results of the neutrality tests are consistent with demographic processes and suggest growth in E. lucens and equilibrium or shrinkage in E. vallentini. Although purifying selection cannot be ruled out in the former, the very few haplotypes recovered from E. vallentini could indicate that any population expansion following a crash is not yet reflected in the relatively slowly evolving mtDNA markers used here. Further work using other methods is recommended.Web of Scienc

The Reinforcing Therapist Performance (RTP) experiment: Study protocol for a cluster randomized trial

<p>Abstract</p> <p>Background</p> <p>Rewarding provider performance has been recommended by the Institute of Medicine as an approach to improve the quality of treatment, yet little empirical research currently exists that has examined the effectiveness and cost-effectiveness of such approaches. The aim of this study is to test the effectiveness and cost-effectiveness of providing monetary incentives directly to therapists as a method to improve substance abuse treatment service delivery and subsequent client treatment outcomes.</p> <p>Design</p> <p>Using a cluster randomized design, substance abuse treatment therapists from across 29 sites were assigned by site to either an implementation as usual (IAU) or pay-for-performance (P4P) condition.</p> <p>Participants</p> <p>Substance abuse treatment therapists participating in a large dissemination and implementation initiative funded by the Center for Substance Abuse Treatment.</p> <p>Intervention</p> <p>Therapists in both conditions received comprehensive training and ongoing monitoring, coaching, and feedback. However, those in the P4P condition also were given the opportunity to earn monetary incentives for achieving two sets of measurable behaviors related to quality implementation of the treatment.</p> <p>Outcomes</p> <p>Effectiveness outcomes will focus on the impact of the monetary incentives to increase the proportion of adolescents who receive a targeted threshold level of treatment, months that therapists demonstrate monthly competency, and adolescents who are in recovery following treatment. Similarly, cost-effectiveness outcomes will focus on cost per adolescent receiving targeted threshold level of treatment, cost per month of demonstrated competence, and cost per adolescent in recovery.</p> <p>Trial Registration</p> <p>Trial Registration Number: NCT01016704</p

Novel Physical Therapy Protocol Results in Increased Compressive Strain and Improved Outcomes in Insertional Achilles Tendinopathy

Category: Ankle, Hindfoot, Sports Introduction/Purpose: Insertional Achilles Tendinopathy (IAT) affects 5% of the general population and up to 20% of the athletic population. Despite trials of non-surgical management, over 50% of patients ultimately pursue surgery. Previously in healthy controls it was demonstrated that ankle dorsiflexion tasks increase transverse compressive strain (force perpendicular to the tendon fibers) most in the deep region of the tendon insertion, where IAT is often most severe. Thus the purpose of this study was to utilize a novel physical therapy protocol that combines isometric and eccentric exercises in order to increase transverse compressive strain and decrease axial tensile strain (force parallel to tendon fibers) in patients with IAT and to determine whether this corresponds with improved patient outcome scores. Methods: Forty-two patients with IAT were enrolled in the study from May 2014 to June 2016, of which twenty-seven patients (mean age: 56.7 + 9.9 years, BMI: 29.6 + 5.9, 56% women) completed the study. A subset (n=15, mean age 58.9 + 8.6 years, BMI: 30.0 + 4.0, 47% women) underwent ultrasound elastography to determine transverse compressive and axial tensile strain in the Achilles tendon during dorsiflexion tasks. Patients were then placed on a physical therapy protocol that focused on progressive loading of the Achilles tendon while avoiding ankle dorsiflexion. Seated isometric plantar flexion, bilateral eccentric heel lowering, and single limb heel lowering exercises were utilized. Questionnaires validated for use in Achilles tendinopathy, the Victorian Institute of Sport Assessment – Achilles (VISA-A) and the Foot and Ankle Ability Measure (FAAM), were completed at the beginning and at the completion of the study. Results: Twenty-three of 27 subjects that completed the study, had clinically significant improvements in their VISA-A (mean change 19.3) or FAAM ADL and sports scores (mean change 16.2 and 22.6, respectively). Ultrasound elastography revealed that the deep region of the Achilles tendon experienced more transverse compressive strain and less axial tensile strain compared to the superficial portion of the tendon when standing. Completion of the physical therapy program resulted in increased transverse compressive strain in the superficial Achilles tendon compared to the pre-therapy value when standing (mean change 52%, p=0.043). Moreover, there was a decrease in axial tensile strain within the deep portion of the tendon in response to physical therapy (mean change 53% p=0.0434). Conclusion: Treatment of IAT patients with a physical therapy protocol utilizing a combination of isometric and eccentric exercises results in improved outcomes, as measured by VISA-A and FAAM questionnaires. Furthermore, ultrasound elastography suggests that while the physical therapy protocol increases the transverse compressive strain in the superficial portion of the Achilles tendon, it results in decreased axial tensile strain in the deep portion of the tendon. Therefore, it is likely that the combination of these two exercise modalities result in the improved clinical outcomes observed in our patients with IAT after undergoing this physical therapy protocol