Comment on 'Accumbens cholinergic interneurons dynamically promote dopamine release and enable motivation'.

Acetylcholine is widely believed to modulate the release of dopamine in the striatum of mammals. Experiments in brain slices clearly show that synchronous activation of striatal cholinergic interneurons is sufficient to drive dopamine release via axo-axonal stimulation of nicotinic acetylcholine receptors. However, evidence for this mechanism in vivo has been less forthcoming. Mohebi, Collins and Berke recently reported that, in awake behaving rats, optogenetic activation of striatal cholinergic interneurons with blue light readily evokes dopamine release measured with the red fluorescent sensor RdLight1 (Mohebi et al., 2023). Here, we show that blue light alone alters the fluorescent properties of RdLight1 in a manner that may be misconstrued as phasic dopamine release, and that this artefactual photoactivation can account for the effects attributed to cholinergic interneurons. Our findings indicate that measurements of dopamine using the red-shifted fluorescent sensor RdLight1 should be interpreted with caution when combined with optogenetics. In light of this and other publications that did not observe large acetylcholine-evoked dopamine transients in vivo, the conditions under which such release occurs in behaving animals remain unknown.

Mesolimbic dopamine ramps reflect environmental timescales.

Mesolimbic dopamine activity occasionally exhibits ramping dynamics, reigniting debate on theories of dopamine signaling. This debate is ongoing partly because the experimental conditions under which dopamine ramps emerge remain poorly understood. Here, we show that during Pavlovian and instrumental conditioning, mesolimbic dopamine ramps are only observed when the inter-trial interval is short relative to the trial period. These results constrain theories of dopamine signaling and identify a critical variable determining the emergence of dopamine ramps.

Dopamine transients follow a striatal gradient of reward time horizons.

Animals make predictions to guide their behavior and update those predictions through experience. Transient increases in dopamine (DA) are thought to be critical signals for updating predictions. However, it is unclear how this mechanism handles a wide range of behavioral timescales-from seconds or less (for example, if singing a song) to potentially hours or more (for example, if hunting for food). Here we report that DA transients in distinct rat striatal subregions convey prediction errors based on distinct time horizons. DA dynamics systematically accelerated from ventral to dorsomedial to dorsolateral striatum, in the tempo of spontaneous fluctuations, the temporal integration of prior rewards and the discounting of future rewards. This spectrum of timescales for evaluative computations can help achieve efficient learning and adaptive motivation for a broad range of behaviors.

Comment on 'Accumbens cholinergic interneurons dynamically promote dopamine release and enable motivation'.

Acetylcholine is widely believed to modulate the release of dopamine in the striatum of mammals. Experiments in brain slices clearly show that synchronous activation of striatal cholinergic interneurons is sufficient to drive dopamine release via axo-axonal stimulation of nicotinic acetylcholine receptors. However, evidence for this mechanism in vivo has been less forthcoming. A recent paper in eLife (Mohebi et al., 2023) reported that, in awake behaving rats, optogenetic activation of striatal cholinergic interneurons with blue light readily evokes dopamine release measured with the red fluorescent sensor RdLight1. Here, we show that blue light alone alters the fluorescent properties of RdLight1 in a manner that may be misconstrued as phasic dopamine release, and that this artefactual photoactivation can account for the effects attributed to cholinergic interneurons. Our findings indicate that measurements of dopamine using the red-shifted fluorescent sensor RdLight1 should be interpreted with caution when combined with optogenetics. In light of this and other publications that did not observe large acetylcholine-evoked dopamine transients in vivo, the conditions under which such release occurs in behaving animals remain unknown.

Lights, fiber, action! A primer on in vivo fiber photometry.

Fiber photometry is a key technique for characterizing brain-behavior relationships in vivo. Initially, it was primarily used to report calcium dynamics as a proxy for neural activity via genetically encoded indicators. This generated new insights into brain functions including movement, memory, and motivation at the level of defined circuits and cell types. Recently, the opportunity for discovery with fiber photometry has exploded with the development of an extensive range of fluorescent sensors for biomolecules including neuromodulators and peptides that were previously inaccessible in vivo. This critical advance, combined with the new availability of affordable "plug-and-play" recording systems, has made monitoring molecules with high spatiotemporal precision during behavior highly accessible. However, while opening exciting new avenues for research, the rapid expansion in fiber photometry applications has occurred without coordination or consensus on best practices. Here, we provide a comprehensive guide to help end-users execute, analyze, and suitably interpret fiber photometry studies.

Selective encoding of reward predictions and prediction errors by globus pallidus subpopulations.

Basal ganglia (BG) circuits help guide and invigorate actions using predictions of future rewards (values). Within the BG, the globus pallidus pars externa (GPe) may play an essential role in aggregating and distributing value information. We recorded from the GPe in unrestrained rats performing both Pavlovian and instrumental tasks to obtain rewards and distinguished neuronal subtypes by their firing properties across the wake/sleep cycle and optogenetic tagging. In both tasks, the parvalbumin-positive (PV+), faster-firing "prototypical" neurons showed strong, sustained modulation by value, unlike other subtypes, including the "arkypallidal" cells that project back to striatum. Furthermore, we discovered that a distinct minority (7%) of GP cells display slower, pacemaker-like firing and encode reward prediction errors (RPEs) almost identically to midbrain dopamine neurons. These cell-specific forms of GPe value representation help define the circuit mechanisms by which the BG contribute to motivation and reinforcement learning.

Accumbens cholinergic interneurons dynamically promote dopamine release and enable motivation.

Motivation to work for potential rewards is critically dependent on dopamine (DA) in the nucleus accumbens (NAc). DA release from NAc axons can be controlled by at least two distinct mechanisms: (1) action potentials propagating from DA cell bodies in the ventral tegmental area (VTA), and (2) activation of ß2* nicotinic receptors by local cholinergic interneurons (CINs). How CIN activity contributes to NAc DA dynamics in behaving animals is not well understood. We monitored DA release in the NAc Core of awake, unrestrained rats using the DA sensor RdLight1, while simultaneously monitoring or manipulating CIN activity at the same location. CIN stimulation rapidly evoked DA release, and in contrast to slice preparations, this DA release showed no indication of short-term depression or receptor desensitization. The sound of unexpected food delivery evoked a brief joint increase in CIN population activity and DA release, with a second joint increase as rats approached the food. In an operant task, we observed fast ramps in CIN activity during approach behaviors, either to start the trial or to collect rewards. These CIN ramps co-occurred with DA release ramps, without corresponding changes in the firing of lateral VTA DA neurons. Finally, we examined the effects of blocking CIN influence over DA release through local NAc infusion of DHßE, a selective antagonist of ß2* nicotinic receptors. DHßE dose-dependently interfered with motivated approach decisions, mimicking the effects of a DA antagonist. Our results support a key influence of CINs over motivated behavior via the local regulation of DA release.

Phasor map analysis to investigate Hutchinson-Gilford progeria cell under polarization-resolved optical scanning microscopy.

Polarized light scanning microscopy is a non-invasive and contrast-enhancing technique to investigate anisotropic specimens and chiral organizations. However, such arrangements suffer from insensitivity to confined blend of structures at sub-diffraction level. Here for the first time, we present that the pixel-by-pixel polarization modulation converted to an image phasor approach issues an insightful view of cells to distinguish anomalous subcellular organizations. To this target, we propose an innovative robust way for identifying changes in the chromatin compaction and distortion of nucleus morphology induced by the activation of the lamin-A gene from Hutchinson-Gilford progeria syndrome that induces a strong polarization response. The phasor mapping is evaluated based on the modulation and phase image acquired from a scanning microscope compared to a confocal fluorescence modality of normal cell opposed to the progeria. The method is validated by characterizing polarization response of starch crystalline granules. Additionally, we show that the conversion of the polarization-resolved images into the phasor could further utilized for segmenting specific structures presenting various optical properties under the polarized light. In summary, image phasor analysis offers a distinctly sensitive fast and easy representation of the polarimetric contrast that can pave the way for remote diagnosis of pathological tissues in real-time.

Combined approach using circular intensity differential scattering microscopy under phasor map data analysis.

Circular intensity differential scattering (CIDS) is based on the analysis of circular polarized light scattering and has been proven to be an interesting label-free microscopy technique sensitive to the chiral organization at the submicroscopic level. However, this approach averages the localized contrasts related to the sample polarimetric properties in the illumination volume. Additionally, the detection sensitivity suffers from the confinement of the mixture of structures, and it becomes an arduous task to discriminate the source of the signal. In this work, we show that a phasor map approach combined with CIDS microscopy has provided an intuitive view of the sample organization to recognize the presence of different molecular species in the illumination volume. The data represented in terms of polarization response mapped to a single point called a phasor also have the potential to pave the way for the analysis of large data sets. We validated this method by numerical simulations and compared the results with that of experimental data of optical devices of reference.

Marked point process representation of oscillatory dynamics underlying working memory.

Objective.Computational models of neural activity at the meso-scale suggest the involvement of discrete oscillatory bursts as constructs of cognitive processing during behavioral tasks. Classical signal processing techniques that attempt to infer neural correlates of behavior from meso-scale activity employ spectral representations of the signal, exploiting power spectral density techniques and time-frequency (T-F) energy distributions to capture band power features. However, such analyses demand more specialized methods that incorporate explicitly the concepts of neurophysiological signal generation and time resolution in the tens of milliseconds. This paper focuses on working memory (WM), a complex cognitive process involved in encoding, storing and retrieving sensory information, which has been shown to be characterized by oscillatory bursts in the beta and gamma band. Employing a generative model for oscillatory dynamics, we present a marked point process (MPP) representation of bursts during memory creation and readout. We show that the markers of the point process quantify specific neural correlates of WM.Approach.We demonstrate our results on field potentials recorded from the prelimbic and secondary motor cortices of three rats while performing a WM task. The generative model for single channel, band-passed traces of field potentials characterizes with high-resolution, the timings and amplitudes of transient neuromodulations in the high gamma (80-150 Hz,γ) and beta (10-30 Hz,ß) bands as an MPP. We use standard hypothesis testing methods on the MPP features to check for significance in encoding of task variables, sensory stimulus and executive control while comparing encoding capabilities of our model with other T-F methods.Main Results.Firstly, the advantages of an MPP approach in deciphering encoding mechanisms at the meso-scale is demonstrated. Secondly, the nature of state encoding by neuromodulatory events is determined. Third, we demonstrate the necessity of a higher time resolution alternative to conventionally employed T-F methods. Finally, our results underscore the novelty in interpreting oscillatory dynamics encompassed by the marked features of the point process.Significance.An MPP representation of meso-scale activity not just enables a rich, high-resolution parameter space for analysis but also presents a novel tool for diverse neural applications.

An expanded palette of dopamine sensors for multiplex imaging in vivo.

Genetically encoded dopamine sensors based on green fluorescent protein (GFP) enable high-resolution imaging of dopamine dynamics in behaving animals. However, these GFP-based variants cannot be readily combined with commonly used optical sensors and actuators, due to spectral overlap. We therefore engineered red-shifted variants of dopamine sensors called RdLight1, based on mApple. RdLight1 can be combined with GFP-based sensors with minimal interference and shows high photostability, permitting prolonged continuous imaging. We demonstrate the utility of RdLight1 for receptor-specific pharmacological analysis in cell culture, simultaneous assessment of dopamine release and cell-type-specific neuronal activity and simultaneous subsecond monitoring of multiple neurotransmitters in freely behaving rats. Dual-color photometry revealed that dopamine release in the nucleus accumbens evoked by reward-predictive cues is accompanied by a rapid suppression of glutamate release. By enabling multiplexed imaging of dopamine with other circuit components in vivo, RdLight1 opens avenues for understanding many aspects of dopamine biology.

Publisher Correction: Dissociable dopamine dynamics for learning and motivation.

Change history: In this Article, an extraneous label appeared in Fig. 4b, and has been removed in the online version.

Dissociable dopamine dynamics for learning and motivation.

The dopamine projection from ventral tegmental area (VTA) to nucleus accumbens (NAc) is critical for motivation to work for rewards and reward-driven learning. How dopamine supports both functions is unclear. Dopamine cell spiking can encode prediction errors, which are vital learning signals in computational theories of adaptive behaviour. By contrast, dopamine release ramps up as animals approach rewards, mirroring reward expectation. This mismatch might reflect differences in behavioural tasks, slower changes in dopamine cell spiking or spike-independent modulation of dopamine release. Here we compare spiking of identified VTA dopamine cells with NAc dopamine release in the same decision-making task. Cues that indicate an upcoming reward increased both spiking and release. However, NAc core dopamine release also covaried with dynamically evolving reward expectations, without corresponding changes in VTA dopamine cell spiking. Our results suggest a fundamental difference in how dopamine release is regulated to achieve distinct functions: broadcast burst signals promote learning, whereas local control drives motivation.

Monogenic Auto-inflammatory Syndromes: A Review of the Literature.

Auto-inflammatory syndromes are a new group of distinct hereditable disorders characterized by episodes of seemingly unprovoked inflammation (most commonly in skin, joints, gut, and eye), the absence of a high titer of auto-antibodies or auto-reactive T cells, and an inborn error of innate immunity. A narrative literature review was carried out of studies related to auto-inflammatory syndromes to discuss the pathogenesis and clinical manifestation of these syndromes. This review showed that the main monogenic auto-inflammatory syndromes are familial Mediterranean fever (FMF), mevalonate kinase deficiency (MKD), Blau syndrome, TNF receptor-associated periodic syndrome (TRAPS), cryopyrin-associated periodic syndrome (CAPS), and pyogenic arthritis with pyoderma gangrenosum and acne (PAPA). The data suggest that correct diagnosis and treatment of monogenic auto-inflammatory diseases relies on the physicians' awareness. Therefore, understanding of the underlying pathogenic mechanisms of auto-inflammatory syndromes, and especially the fact that these disorders are mediated by IL-1 secretion stimulated by monocytes and macrophages, facilitated significant progress in patient management.

Is systolic blood pressure below 150 mm Hg an appropriate goal for primary prevention of cardiovascular events among elderly population?

Recently, Joint National Committee has changed the optimal therapeutic goal of systolic blood pressure (SBP) up to 150 mm Hg for elderly population. We aimed to investigate impact of different blood pressure (BP) categories on risk of developing cardiovascular disease (CVD) and mortality among elderly. The present study included 1845 participants, aged ≥60 years (mean age = 65 years), free of CVD at baseline, who had undergone health examinations between January 1999 and 2001, and were followed up until March 2010. Cox proportional hazard regression was performed to assess the hazard ratios (HRs) of BP categories for CVD and mortality events, considering those with optimal BP (SBP <120 mm Hg and diastolic BP [DBP] <80 mm Hg) as reference. During a median of 10 years follow-up, 380 cases of first CVD and 260 cases of mortality events occurred. In multivariable adjusted model, prehypertensive group (SBP between 120-129 mm Hg or DBP between 80-85 mm Hg) could not predict CVD (HR, 0.87 [0.61-1.24]) nor mortality events (HR, 0.86 [0.58-1.34]). Those with SBP between 140 mm Hg and 150 mm Hg (group 3) were at higher risk for developing CVD (HR, 1.79 [1.17-2.74]), but there were no significant risk for total mortality (HR, 1.13 [0.65-1.97]). Hypertensive group (SBP ≥150 mm Hg or DBP ≥90 mm Hg or taking antihypertensive drugs) was associated with increased risk of both CVD (HR, 1.73 [1.24-2.42]) and mortality events (HR, 1.49 [1.00-2.23]).However, Joint National Committee 8 suggested no more benefit with lowering SBP <150 mm Hg, but the results of this study imply that those with SBP between 150 mm Hg and 140 mm Hg are still at elevated risk for CVD/coronary heart disease events.

A fully automated rodent conditioning protocol for sensorimotor integration and cognitive control experiments.

Rodents have been traditionally used as a standard animal model in laboratory experiments involving a myriad of sensory, cognitive, and motor tasks. Higher cognitive functions that require precise control over sensorimotor responses such as decision-making and attentional modulation, however, are typically assessed in nonhuman primates. Despite the richness of primate behavior that allows multiple variants of these functions to be studied, the rodent model remains an attractive, cost-effective alternative to primate models. Furthermore, the ability to fully automate operant conditioning in rodents adds unique advantages over the labor intensive training of nonhuman primates while studying a broad range of these complex functions. Here, we introduce a protocol for operantly conditioning rats on performing working memory tasks. During critical epochs of the task, the protocol ensures that the animal's overt movement is minimized by requiring the animal to 'fixate' until a Go cue is delivered, akin to nonhuman primate experimental design. A simple two alternative forced choice task is implemented to demonstrate the performance. We discuss the application of this paradigm to other tasks.

Sorting and tracking neuronal spikes via simple thresholding.

A fundamental goal in systems neuroscience is to assess the individual as well as the synergistic roles of single neurons in a recorded ensemble as they relate to an observed behavior. A mandatory step to achieve this goal is to sort spikes in an extracellularly recorded mixture that belong to individual neurons through feature extraction and clustering techniques. Here, we propose an approach for approximating the often nonlinear and time varying decision boundaries between spike-derived feature classes based on a simple, yet optimal thresholding mechanism. Because thresholding is a binary classifier, we show that the complex nonlinear decision boundaries required for spike class discrimination can be achieved by adequately fusing a set of weak binary classifiers. The thresholds for these binary classifiers are adaptively estimated through a learning algorithm that maximizes the separability between the sparsely represented classes. Based on our previous work, the approach substantially reduces the computational complexity of extracting, aligning and sorting multiple single unit activity early in the data stream. Here, we also show its ability to track changes in spike features over extended periods of time, making it highly suitable for basic neuroscience studies as well as for implementation in miniaturized, fully implantable electronics in brain-machine interface applications.

Hydatid cyst of the parotid gland.

Primary hydatid cyst of the parotid gland is extremely rare, even in the endemic areas. A 23-year-old woman presented with slowly progressive swelling in the right periauricular region. Computed tomography (CT) scan of the head and neck revealed a round, well-demarcated water-density mass in the right parotid gland. At the operation, the cystic mass replacing most of the superficial part of right parotid gland was demonstrated. Superficial parotidectomy was carried out. Histopathological examination confirmed the diagnosis of hydatid disease. CT scan is a valuable imaging method for diagnosis of parotid cystic lesions; however, other acquired and congenital cystic lesions of parotid gland may have similar appearance and should be differentiated. Where the incidence of the disease is high, hydatid cyst of parotid gland should be considered in the differential diagnosis of lesions causing swelling of the parotid area.