A lipid-anchored neurokinin 1 receptor antagonist prolongs pain relief by a three-pronged mechanism of action targeting the receptor at the plasma membrane and in endosomes

G-protein-coupled receptors (GPCRs) are traditionally known for signaling at the plasma membrane, but they can also signal from endosomes after internalization to control important pathophysiological processes. In spinal neurons, sustained endosomal signaling of the neurokinin 1 receptor (NK1R) mediates nociception, as demonstrated in models of acute and neuropathic pain. An NK1R antagonist, Spantide I (Span), conjugated to cholestanol (Span-Chol), accumulates in endosomes, inhibits endosomal NK1R signaling, and causes prolonged antinociception. However, the extent to which the Chol-anchor influences long-term location and activity is poorly understood. Herein, we used fluorescent correlation spectroscopy and targeted biosensors to characterize Span-Chol over time. The Chol-anchor increased local concentration of probe at the plasma membrane. Over time we observed an increase in NK1R-binding affinity and more potent inhibition of NK1R-mediated calcium signaling. Span-Chol, but not Span, caused a persistent decrease in NK1R recruitment of βarrestin and receptor internalization to early endosomes. Using targeted biosensors, we mapped the relative inhibition of NK1R signaling as the receptor moved into the cell. Span selectively inhibited cell surface signaling, whereas Span-Chol partitioned into endosomal membranes and blocked endosomal signaling. In a preclinical model of pain, Span-Chol caused prolonged antinociception (>9 h), which is attributable to a three-pronged mechanism of action: increased local concentration at membranes, a prolonged decrease in NK1R endocytosis, and persistent inhibition of signaling from endosomes. Identifying the mechanisms that contribute to the increased preclinical efficacy of lipid-anchored NK1R antagonists is an important step toward understanding how we can effectively target intracellular GPCRs in disease

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

brainlife.io: a decentralized and open-source cloud platform to support neuroscience research

Neuroscience is advancing standardization and tool development to support rigor and transparency. Consequently, data pipeline complexity has increased, hindering FAIR (findable, accessible, interoperable and reusable) access. brainlife.io was developed to democratize neuroimaging research. The platform provides data standardization, management, visualization and processing and automatically tracks the provenance history of thousands of data objects. Here, brainlife.io is described and evaluated for validity, reliability, reproducibility, replicability and scientific utility using four data modalities and 3,200 participants

credit, including © notice, is given to the source. Religious Identity and Economic Behavior

We thank Azim Shariff and Ara Norenzayan for sharing with us the religious identity priming instrument. We are grateful to Stefano DellaVigna, Kirabo Jackson, Ted O’Donoghue, and participants at the NBER’

Neurokinin 1 receptor signaling in endosomes mediates sustained nociception and is a viable therapeutic target for prolonged pain relief

International audienceTypically considered to be cell surface sensors of extracellular signals, heterotrimeric GTP-binding protein (G protein)-coupled receptors (GPCRs) control many pathophysiological processes and are the target of 30% of therapeutic drugs. Activated receptors redistribute to endosomes, but researchers have yet to explore whether endosomal receptors generate signals that control complex processes in vivo and are viable therapeutic targets. We report that the substance P (SP) neurokinin 1 receptor (NK1R) signals from endosomes to induce sustained excitation of spinal neurons and pain transmission and that specific antagonism of the NK1R in endosomes with membrane-anchored drug conjugates providesmore effective and sustained pain relief than conventional plasmamembrane-targeted antagonists. Pharmacological and genetic disruption of clathrin, dynamin, and b-arrestin blocked SP-induced NK1R endocytosis and prevented SP-stimulated activation of cytosolic protein kinase C and nuclear extracellular signal-regulated kinase, as well as transcription. Endocytosis inhibitors prevented sustained SP-induced excitation of neurons in spinal cord slices in vitro and attenuated nociception in vivo. When conjugated to cholestanol to promote endosomal targeting, NK1R antagonists selectively inhibited endosomal signaling and sustained neuronal excitation. Cholestanol conjugation amplified and prolonged the antinociceptive actions of NK1R antagonists. These results reveal a critical role for endosomal signaling of the NK1R in the complex pathophysiology of pain and demonstrate the use of endosomally targeted GPCR antagonists

The Power and Pitfalls of Experiments in Development Economics: Some Non-random Reflections

Impact evaluation based on randomized controlled trials (RCTs) offers a powerful tool that has fundamentally reshaped development economics by offering novel solutions to long-standing problems of weak causal identification. Nonetheless, RCTs suffer important and underappreciated pitfalls, some of which are intrinsic to the method when applied to economic problems, others that are the result of methodological boosterism. Among the pitfalls are ethical dilemmas, uncontrollable treatments that result in a 'faux exogeneity,' distortion of the research agenda, and a tendency to estimate interventions' abstract efficacy rather than their effectiveness in practice. We illustrate these points through the literature on smallholder capital access and productivity growth. Ultimately, we argue for a methodological pluralism that recognizes all identification strategies' limitations. Copyright 2010, Oxford University Press.

Beyond Point Masses. II. Non-Keplerian Shape Effects Are Detectable in Several TNO Binaries

About 40 trans-Neptunian binaries (TNBs) have fully determined orbits with about 10 others being solved except for breaking the mirror ambiguity. Despite decades of study, almost all TNBs have only ever been analyzed with a model that assumes perfect Keplerian motion (e.g., two point masses). In reality, all TNB systems are non-Keplerian due to nonspherical shapes, possible presence of undetected system components, and/or solar perturbations. In this work, we focus on identifying candidates for detectable non-Keplerian motion based on sample of 45 well-characterized binaries. We use MultiMoon , a non-Keplerian Bayesian inference tool, to analyze published relative astrometry allowing for nonspherical shapes of each TNB system’s primary. We first reproduce the results of previous Keplerian fitting efforts with MultiMoon , which serves as a comparison for the non-Keplerian fits and confirms that these fits are not biased by the assumption of a Keplerian orbit. We unambiguously detect non-Keplerian motion in eight TNB systems across a range of primary radii, mutual orbit separations, and system masses. As a proof of concept for non-Keplerian fitting, we perform detailed fits for (66652) Borasisi-Pabu, possibly revealing a J _2 ≈ 0.44, implying Borasisi (and/or Pabu) may be a contact binary or an unresolved compact binary. However, full confirmation of this result will require new observations. This work begins the next generation of TNB analyses that go beyond the point mass assumption to provide unique and valuable information on the physical properties of TNBs with implications for their formation and evolution