Upstream Circuits Research Articles

Using the MouseWalker to Quantify Locomotor Dysfunction in a Mouse Model of Spinal Cord Injury.

The execution of complex and highly coordinated motor programs, such as walking and running, is dependent on the rhythmic activation of spinal and supra-spinal circuits. After a thoracic spinal cord injury, communication with upstream circuits is impaired. This, in turn, leads to a loss of coordination, with limited recovery potential. Hence, to better evaluate the degree of recovery after the administration of drugs or therapies, there is a necessity for new, more detailed, and accurate tools to quantify gait, limb coordination, and other fine aspects of locomotor behavior in animal models of spinal cord injury. Several assays have been developed over the years to quantitatively assess free-walking behavior in rodents; however, they usually lack direct measurements related to stepping gait strategies, footprint patterns, and coordination. To address these shortcomings, an updated version of the MouseWalker, which combines a frustrated total internal reflection (fTIR) walkway with tracking and quantification software, is provided. This open-source system has been adapted to extract several graphical outputs and kinematic parameters, and a set of post-quantification tools can be to analyze the output data provided. This manuscript also demonstrates how this method, allied with already established behavioral tests, quantitatively describes locomotor deficits following spinal cord injury.

Temporal scaling and computing time in neural circuits: Should we stop watching the clock and look for its gears?

Timing underlies a variety of functions, from walking to perceiving causality. Neural timing models typically fall into one of two categories-"ramping" and "population-clock" theories. According to ramping models, individual neurons track time by gradually increasing or decreasing their activity as an event approaches. To time different intervals, ramping neurons adjust their slopes, ramping steeply for short intervals and vice versa. In contrast, according to "population-clock" models, multiple neurons track time as a group, and each neuron can fire nonlinearly. As each neuron changes its rate at each point in time, a distinct pattern of activity emerges across the population. To time different intervals, the brain learns the population patterns that coincide with key events. Both model categories have empirical support. However, they often differ in plausibility when applied to certain behavioral effects. Specifically, behavioral data indicate that the timing system has a rich computational capacity, allowing observers to spontaneously compute novel intervals from previously learned ones. In population-clock theories, population patterns map to time arbitrarily, making it difficult to explain how different patterns can be computationally combined. Ramping models are viewed as more plausible, assuming upstream circuits can set the slope of ramping neurons according to a given computation. Critically, recent studies suggest that neurons with nonlinear firing profiles often scale to time different intervals-compressing for shorter intervals and stretching for longer ones. This "temporal scaling" effect has led to a hybrid-theory where, like a population-clock model, population patterns encode time, yet like a ramping neuron adjusting its slope, the speed of each neuron's firing adapts to different intervals. Here, we argue that these "relative" population-clock models are as computationally plausible as ramping theories, viewing population-speed and ramp-slope adjustments as equivalent. Therefore, we view identifying these "speed-control" circuits as a key direction for evaluating how the timing system performs computations. Furthermore, temporal scaling highlights that a key distinction between different neural models is whether they propose an absolute or relative time-representation. However, we note that several behavioral studies suggest the brain processes both scales, cautioning against a dichotomy.

Sleep-promoting neurons remodel their response properties to calibrate sleep drive with environmental demands.

Falling asleep at the wrong time can place an individual at risk of immediate physical harm. However, not sleeping degrades cognition and adaptive behavior. To understand how animals match sleep need with environmental demands, we used live-brain imaging to examine the physiological response properties of the dorsal fan-shaped body (dFB) following interventions that modify sleep (sleep deprivation, starvation, time-restricted feeding, memory consolidation) in Drosophila. We report that dFB neurons change their physiological response-properties to dopamine (DA) and allatostatin-A (AstA) in response to different types of waking. That is, dFB neurons are not simply passive components of a hard-wired circuit. Rather, the dFB neurons intrinsically regulate their response to the activity from upstream circuits. Finally, we show that the dFB appears to contain a memory trace of prior exposure to metabolic challenges induced by starvation or time-restricted feeding. Together, these data highlight that the sleep homeostat is plastic and suggests an underlying mechanism.

Organization of the gravity-sensing system in zebrafish

Motor circuits develop in sequence from those governing fast movements to those governing slow. Here we examine whether upstream sensory circuits are organized by similar principles. Using serial-section electron microscopy in larval zebrafish, we generated a complete map of the gravity-sensing (utricular) system spanning from the inner ear to the brainstem. We find that both sensory tuning and developmental sequence are organizing principles of vestibular topography. Patterned rostrocaudal innervation from hair cells to afferents creates an anatomically inferred directional tuning map in the utricular ganglion, forming segregated pathways for rostral and caudal tilt. Furthermore, the mediolateral axis of the ganglion is linked to both developmental sequence and neuronal temporal dynamics. Early-born pathways carrying phasic information preferentially excite fast escape circuits, whereas later-born pathways carrying tonic signals excite slower postural and oculomotor circuits. These results demonstrate that vestibular circuits are organized by tuning direction and dynamics, aligning them with downstream motor circuits and behaviors.

Unified classification of mouse retinal ganglion cells using function, morphology, and gene expression.

SUMMARYClassification and characterization of neuronal types are critical for understanding their function and dysfunction. Neuronal classification schemes typically rely on measurements of electrophysiological, morphological, and molecular features, but aligning such datasets has been challenging. Here, we present a unified classification of mouse retinal ganglion cells (RGCs), the sole retinal output neurons. We use visually evoked responses to classify 1,859 mouse RGCs into 42 types. We also obtain morphological or transcriptomic data from subsets and use these measurements to align the functional classification to publicly available morphological and transcriptomic datasets. We create an online database that allows users to browse or download the data and to classify RGCs from their light responses using a machine learning algorithm. This work provides a resource for studies of RGCs, their upstream circuits in the retina, and their projections in the brain, and establishes a framework for future efforts in neuronal classification and open data distribution.

Prefrontal Input to the Periaqueductal Gray Controls Placebo Analgesia

Placebo analgesia is associated with activity in a highly interconnected neural network distributed throughout the brain. However, this is based on a large body of functional imaging work that primarily relies on correlative measures of neural activity and neurochemical signaling obtained in human subjects, making it challenging to ascribe causal roles to the underlying neural pathways. To address this gap, we developed a mouse model of placebo analgesia and used chemogenetic loss-of-function manipulations to identify the underlying neural circuits. We specifically addressed the role of the descending pain modulatory system (DPMS) and its upstream cortical circuits. We found that chemogenetic inactivation of a subpopulation of excitatory neurons in the ventrolateral periaqueductal gray (vlPAG) abolishes both morphine analgesia and morphine-conditioned placebo analgesia. Placebo, but not morphine analgesia, depends on vlPAG-projecting neurons in the medial prefrontal cortex (mPFC) and anterior cingulate cortex (ACC), but not the anterior insula (AI). For placebo to occur, mPFC-to-vlPAG neurons must be active during conditioning, consistent with a critical role for activity-dependent plasticity in this pathway. Our findings suggest that the vlPAG implements cognitive pain modulation by integrating top-down commands, at least some of which are conveyed directly from the cortex. Grant support from Rita Allen Foundation (non-profit) NIDA grant (R00 DA 034648) Klingenstein-Simons Foundation (non-profit).

Activity in projection neurons from prelimbic cortex to the PVT is necessary for retrieval of morphine withdrawal memory.

Previous work has shown that the paraventricular nucleus of the thalamus (PVT) is an important region that is involved in the conditioned context-induced retrieval of morphine withdrawal memory. However, the upstream neural circuits that activate the PVT to participate in the conditioned context-induced retrieval of morphine withdrawal memory remain unknown. In the present work, we find that the conditioned context activates projection neurons from the prelimbic cortex (PrL) to the PVT, and the inhibition of PrL-PVT projection neurons inhibits the conditioned context-induced retrieval of morphine withdrawal memory; the conditioned context induces an increase in Arc expression, intrinsic excitability, and glutamate output in PrL-PVT projection neurons in morphine-withdrawn mice. These results suggest that the activity of PrL-PVT projection neurons is necessary for the retrieval of morphine withdrawal memory, and the conditioned context causes a plastic change in the activity in these projection neurons during the withdrawal memory retrieval.

Dysregulation of Midbrain Dopamine System and the Pathophysiology of Schizophrenia.

Dysregulation of the dopamine system is central to many models of the pathophysiology of psychosis in schizophrenia. However, emerging evidence suggests that this dysregulation is driven by the disruption of upstream circuits that provide afferent control of midbrain dopamine neurons. Furthermore, stress can profoundly disrupt this regulatory circuit, particularly when it is presented at critical vulnerable prepubertal time points. This review will discuss the dopamine system and the circuits that regulate it, focusing on the hippocampus, medial prefrontal cortex, thalamic nuclei, and medial septum, and the impact of stress. A greater understanding of the regulation of the dopamine system and its disruption in schizophrenia may provide a more complete neurobiological framework to interpret clinical findings and develop novel treatments.

A Sensitization Of Projection Neurons from Prelimbic Cortex to Paraventricular Nucleus of Thalamus Induced By Retrieval of Morphine Withdrawal Memory

Exposure to context that is associated with drug withdrawal syndromes induces the retrieval of drug withdrawal memory and leads to drug relapse. Previous study showed that the paraventricular nucleus of the thalamus (PVT) was an important region that was involved in conditioned context-induced retrieval of drug withdrawal memory. However, the upstream neuronal circuits that activate the PVT to participate in conditioned context-induced retrieval of morphine withdrawal memory remain to be studied. In the present work, using retrograde labeling method combined with immunohistochemical, chemical genetic, optogenetic and whole-cell recording approaches, we found that that projection neurons from the prelimbic cortex to the PVT played an important role in conditioned context-induced retrieval of morphine withdrawal memory and the retrieval of morphine withdrawal memory induced an increase in intrinsic excitability and glutamate output in PrL-PVT projection neurons in morphine withdrawn mice, suggesting a sensitization of PrL-PVT projection neurons after exposure to conditioned context.

Recurrent architecture for adaptive regulation of learning in the insect brain.

Dopaminergic neurons (DANs) drive learning across the animal kingdom, but the upstream circuits that regulate their activity and thereby learning remain poorly understood. We provide the first synaptic-resolution connectome of the circuitry upstream of all DANs in a learning center, the mush-room body (MB) of Drosophila larva. We discover afferent sensory pathways and a large population of neurons that provide feedback from MB output neurons and link distinct memory systems (aversive and appetitive). We combine this with functional studies of DANs and their presynaptic partners and with comprehensive circuit modelling. We find that DANs compare convergent feedback from aversive and appetitive systems which enables the computation of integrated predictions that may improve future learning. Computational modelling reveals that the discovered feedback motifs increase model flexibility and performance on learning tasks. Our study provides the most detailed view to date of biological circuit motifs that support associative learning.

Metabolic reprogramming as a feast for virus replication.

Viral replication depends entirely on the energy and biosynthetic precursors supplied by the host cell metabolic network. Viruses actively reprogram host cell metabolism to establish optimal environment for their replication and spread. They stimulate the uptake of extracellular nutrients and predominantly modulate glucose, glutamine, and fatty acid metabolism to support anabolic metabolic pathways. Some viruses activate the process of aerobic glycolysis, divert the glycolytic carbon for biosynthetic reactions, and stimulate glutamine utilization to replenish tricarboxylic cycle intermediates. Others use glutamine carbon to promote de novo fatty acid synthesis, amino acid supply or glutathione production. The unique metabolic signature and different dependence of viral life cycle on the individual metabolic processes is therefore characteristic feature of almost each virus. Deeper understanding of how viruses alter cellular metabolic pathways or their upstream regulatory circuits may lead to development of more effective antiviral treatment strategies based on targeted metabolic inhibition. Keywords: virus infection; metabolism; glycolysis; glutamine metabolism; fatty acid synthesis; metabolic reprogramming; virus-host interaction.

A Series Circuit from CA1 of Dorsal Hippocampus to Basolateral Amygdala via Postrhinal Cortex Participates in Conditioned Context-Induced Retrieval of Morphine Withdrawal Memory

Background: Conditioned context-induced retrieval of drug withdrawal memory contributes to drug relapse. The basolateral amygdala (BLA) plays an important role in conditioned context-induced retrieval of morphine withdrawal memory. However, the upstream pathways of the activation of the BLA by conditioned context remains to be studied. Methods: Using the conditioned place aversion model, chemical-genetic method, trans-neuronal virus tracing technique and immunohistochemical method, we studied the upstream neural circuits of the activation of the BLA during conditioned context-induced retrieval of morphine withdrawal memory. Findings: The present results show that the dorsal CA1 of the hippocampus is an upstream brain region of the activation of the BLA during conditioned context-induced withdrawal memory retrieval; the indirect connection from dorsal CA1 of the hippocampus to the BLA is enhanced in mice with conditioned place aversion; the postrhinal cortex (POR) is a brain region that connects the dorsal CA1 of the hippocampus and the activation of the BLA in conditioned context-induced retrieval of morphine withdrawal memory. Interpretation: These results suggest that a series circuit from CA1 of dorsal hippocampus to basolateral amygdala via postrhinal cortex participates in conditioned context-induced retrieval of morphine withdrawal memory. Funding Statement: This study was supported by the National Program of Basic Research sponsored by the Ministry of Science and Technology of China (2009CB52201 and 2013CB835100) and Project of Foundation of National Natural Science of China (31121061, 91332204, 81371466 and 31070932). Declaration of Interests: All authors declare no conflicts of interest. Ethics Approval Statement: All experimental procedures conformed to Fudan University as well as international guidelines on the ethical use of animals. All efforts were made to minimize animal suffering and reduce the number of animals used.

Typology and Circuitry of Suppressed-by-Contrast Retinal Ganglion Cells.

Retinal ganglion cells (RGCs) relay ~40 parallel and independent streams of visual information, each encoding a specific feature of a visual scene, to the brain for further processing. The polarity of a visual neuron’s response to a change in contrast is generally the first characteristic used for functional classification: ON cells increase their spike rate to positive contrast; OFF cells increase their spike rate for negative contrast; ON-OFF cells increase their spike rate for both contrast polarities. Suppressed-by-Contrast (SbC) neurons represent a less well-known fourth category; they decrease firing below a baseline rate for both positive and negative contrasts. SbC RGCs were discovered over 50 years ago, and SbC visual neurons have now been found in the thalamus and primary visual cortex of several mammalian species, including primates. Recent discoveries of SbC RGCs in mice have provided new opportunities for tracing upstream circuits in the retina responsible for the SbC computation and downstream targets in the brain where this information is used. We review and clarify recent work on the circuit mechanism of the SbC computation in these RGCs. Studies of mechanism rely on precisely defined cell types, and we argue that, like ON, OFF, and ON-OFF RGCs, SbC RGCs consist of more than one type. A new appreciation of the diversity of SbC RGCs will help guide future work on their targets in the brain and their roles in visual perception and behavior.

Routes to, from and within the subiculum.

The subiculum is one of the major output areas of the hippocampus and has extensive projections to extrahippocampal targets. It is likely to play a pivotal role in the distribution of outgoing information from the hippocampus. The hippocampus, including the subiculum, is important for the formation, consolidation and retrieval of memory. These functions require a network that is flexible enough to encode incoming information and also allows for reliable distribution, storage and integration into previously encoded memories. Finally, relevant information has to be retrieved in a context-specific manner to allow for an appropriate behavioral response. The subiculum as a gateway between the hippocampus and cortex might serve to integrate and process information from the hippocampus proper and its other inputs before conveying it to more permanent storage locations. This review summarizes how the subiculum is embedded into upstream and downstream circuits, describes what is known about the local network topology and discusses cellular and functional properties of subicular cells subtypes. Lastly, it describes how these properties might help to separate information into parallel output streams and distribute it to its multiple target areas.

Visual Circuits for Direction Selectivity.

Images projected onto the retina of an animal eye are rarely still. Instead, they usually contain motion signals originating either from moving objects or from retinal slip caused by self-motion. Accordingly, motion signals tell the animal in which direction a predator, prey, or the animal itself is moving. At the neural level, visual motion detection has been proposed to extract directional information by a delay-and-compare mechanism, representing a classic example of neural computation. Neurons responding selectively to motion in one but not in the other direction have been identified in many systems, most prominently in the mammalian retina and the fly optic lobe. Technological advances have now allowed researchers to characterize these neurons' upstream circuits in exquisite detail. Focusing on these upstream circuits, we review and compare recent progress in understanding the mechanisms that generate direction selectivity in the early visual system of mammals and flies.

Gap Junction-Mediated Signaling from Motor Neurons Regulates Motor Generation in the Central Circuits of Larval Drosophila.

In this study, we used the peristaltic crawling of Drosophila larvae as a model to study how motor patterns are regulated by central circuits. We built an experimental system that allows simultaneous application of optogenetics and calcium imaging to the isolated ventral nerve cord (VNC). We then investigated the effects of manipulating local activity of motor neurons (MNs) on fictive locomotion observed as waves of MN activity propagating along neuromeres. Optical inhibition of MNs with halorhodopsin3 in a middle segment (A4, A5, or A6), but not other segments, dramatically decreased the frequency of the motor waves. Conversely, local activation of MNs with channelrhodopsin2 in a posterior segment (A6 or A7) increased the frequency of the motor waves. Since peripheral nerves mediating sensory feedback were severed in the VNC preparation, these results indicate that MNs send signals to the central circuits to regulate motor pattern generation. Our results also indicate segmental specificity in the roles of MNs in motor control. The effects of the local MN activity manipulation were lost in shaking-B2 (shakB2 ) or ogre2 , gap-junction mutations in Drosophila, or upon acute application of the gap junction blocker carbenoxolone, implicating electrical synapses in the signaling from MNs. Cell-type-specific RNAi suggested shakB and ogre function in MNs and interneurons, respectively, during the signaling. Our results not only reveal an unexpected role for MNs in motor pattern regulation, but also introduce a powerful experimental system that enables examination of the input-output relationship among the component neurons in this system.SIGNIFICANCE STATEMENT Motor neurons are generally considered passive players in motor pattern generation, simply relaying information from upstream interneuronal circuits to the target muscles. This study shows instead that MNs play active roles in the control of motor generation by conveying information via gap junctions to the central pattern-generating circuits in larval Drosophila, providing novel insights into motor circuit control. The experimental system introduced in this study also presents a new approach for studying intersegmentally coordinated locomotion. Unlike traditional electrophysiology methods, this system enables the simultaneous recording and manipulation of populations of neurons that are genetically specified and span multiple segments.

Motor neurons control locomotor circuit function retrogradely via gap junctions.

Motor neurons are the final stage of neural processing for the execution of motor behaviours. Traditionally, motor neurons have been viewed as the 'final common pathway', serving as passive recipients merely conveying to the muscles the final motor program generated by upstream interneuron circuits. Here we reveal an unforeseen role of motor neurons in controlling the locomotor circuit function via gap junctions in zebrafish. These gap junctions mediate a retrograde analogue propagation of voltage fluctuations from motor neurons to control the synaptic release and recruitment of the upstream V2a interneurons that drive locomotion. Selective inhibition of motor neurons during ongoing locomotion de-recruits V2a interneurons and strongly influences locomotor circuit function. Rather than acting as separate units, gap junctions unite motor neurons and V2a interneurons into functional ensembles endowed with a retrograde analogue computation essential for locomotor rhythm generation. These results show that motor neurons are not a passive recipient of motor commands but an integral component of the neural circuits responsible for motor behaviour.

Automotive Turbochargers Power Estimation Based on Speed Fluctuation Analysis

Turbocharging technology will play a crucial role in the near future as a way to meet the requirements for pollutant emissions and fuel consumption reduction.However, optimal turbocharger control is still an issue, especially for downsized engines fitted with a low number of cylinders. As a matter of fact, automotive turbochargers are characterized by wide operating range and unsteady gas flow through the turbine, while only steady flow maps are usually provided by the manufacturer. In addition, in passenger cars applications, real-time turbocharger optimal control is even more difficult because of the lack of information about pressure/temperature in turbine upstream/downstream circuits and turbocharger rotational speed.In order to overcome these unknowns, this work presents a methodology for instantaneous turbocharger rotational speed determination through a proper processing of the signal coming from one accelerometer mounted on the compressor diffuser, or one microphone facing the compressor. The presented approach can be used to evaluate both turbocharger speed mean value and the amplitude of turbocharger speed fluctuations caused by the pulsating gas flow in turbine upstream and downstream circuits. Once turbocharger speed has been determined, it can be used to estimate power delivered by the turbine.The whole estimation algorithm has been developed and validated for a light duty turbocharged Common-Rail Diesel engine mounted in a test cell. However, the developed methodology is general and can be applied to different turbochargers, both for Spark Ignited and Diesel applications.

Integrating DNA-strand-displacement circuitry with self-assembly of spherical nucleic acids.

Programmable and algorithmic behaviors of DNA molecules allow one to control the structures of DNA-assembled materials with nanometer precision and to construct complex networks with digital and analog behaviors. Here we developed a way of integrating a DNA-strand-displacement circuit with self-assembly of spherical nucleic acids, wherein a single DNA strand was used to initiate and catalyze the operation of upstream circuits to release a single strand that subsequently triggers self-assembly of spherical nucleic acids in downstream circuits, realizing a programmable kinetic control of self-assembly of spherical nucleic acids. Through utilizing this method, single-nucleotide polymorphisms or indels occurring at different positions of a sequence of oligonucleotide were unambiguously discriminated. We provide here a sophisticated way of combining the DNA-strand-displacement-based characteristic of DNA with the distinct assembly properties of inorganic nanoparticles, which may find broad potential applications in the fabrication of a wide range of complex multicomponent devices and architectures.

Estimation Methodology for Automotive Turbochargers Speed Fluctuations Due to Pulsating Flows

Turbocharging technique, together with engine downsizing, will play a fundamental role in the near future as a way to reach the required maximum performance while reducing engine displacement and, consequently, CO2 emissions. However, performing an optimal control of the turbocharging system is very difficult, especially for small engines fitted with a low number of cylinders. This is mainly due to the high turbocharger operating range and to the fact that the flow through compressor and turbine is highly unsteady, while only steady-flow maps are usually provided by the manufacturer. In addition, in passenger cars applications, it is usually difficult to optimize turbocharger operating conditions because of the lack of information about pressure/temperature in turbine upstream/downstream circuits and turbocharger rotational speed. This work presents a methodology suitable for instantaneous turbocharger rotational speed determination through a proper processing of the signal coming from an accelerometer mounted on the compressor diffuser or a microphone faced to the compressor. The presented approach can be used to evaluate turbocharger speed mean value and turbocharger speed fluctuation (due to unsteady flow in turbine upstream and downstream circuits), which can be correlated to the power delivered by the turbine. The whole estimation algorithm has been developed and validated for a light-duty turbocharged common-rail diesel engine mounted in a test cell. Nevertheless, the developed methodology is general and can be applied to different turbochargers, both for spark ignited and diesel applications.