A genetically encoded tool for reconstituting synthetic modulatory neurotransmission and reconnect neural circuits in vivo

© The Author(s), 2021. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Hawk, J. D., Wisdom, E. M., Sengupta, T., Kashlan, Z. D., & Colon-Ramos, D. A. A genetically encoded tool for reconstituting synthetic modulatory neurotransmission and reconnect neural circuits in vivo. Nature Communications, 12(1), (2021): 4795, https://doi.org/10.1038/s41467-021-24690-9.Chemogenetic and optogenetic tools have transformed the field of neuroscience by facilitating the examination and manipulation of existing circuits. Yet, the field lacks tools that enable rational rewiring of circuits via the creation or modification of synaptic relationships. Here we report the development of HySyn, a system designed to reconnect neural circuits in vivo by reconstituting synthetic modulatory neurotransmission. We demonstrate that genetically targeted expression of the two HySyn components, a Hydra-derived neuropeptide and its receptor, creates de novo neuromodulatory transmission in a mammalian neuronal tissue culture model and functionally rewires a behavioral circuit in vivo in the nematode Caenorhabditis elegans. HySyn can interface with existing optogenetic, chemogenetic and pharmacological approaches to functionally probe synaptic transmission, dissect neuropeptide signaling, or achieve targeted modulation of specific neural circuits and behaviors.This work was initiated in the Grass Laboratory at the Marine Biological Laboratories (MBL) with funding through a Grass Fellowship awarded to J.D.H. Thanks to Richard Goodman (OHSU) for encouragement during the conceptualization of the fellowship application, and the 2019 Grass Fellows, Mel Coleman (Grass Director), and Christophe Dupré (Associate Director) for advice and support during the summer fellowship. We thank the MBL Division of Education and participants in the Vendor Equipment Loan Program. Special thanks to Sutter Instruments, who generously provided all electrophysiology equipment and substantial on-site assistance, and Zeiss, who provided on-site assistance at MBL. We thank Zhao-Wen Wang and Ping Liu (UConn) for guidance and training in patch-clamp electrophysiology, as well as providing Neuro2a cells. We thank Rob Steele (UCI) for supplying Hydra, as well as advice and inspiration on Hydra biology. We thank members of the Colón-Ramos lab and Hari Shroff (NIH) for thoughtful comments on the manuscript. We thank Michael Koelle and Andrew Olson (Yale University) for advice and reagents regarding serotonin rewiring experiments. We also thank Steve Flavell (MIT) for ideas and reagents regarding the experiments associated with del-7. We thank Life Science Editors for editing assistance. D.A.C.-R. is an MBL Fellow. Research in the D.A.C.-R. lab was supported by NIH R01NS076558, DP1NS111778, and by an HHMI Scholar Award

Functional Organization of a Neural Network for Aversive Olfactory Learning in Caenorhabditis elegans

Many animals use their olfactory systems to learn to avoid dangers, but how neural circuits encode naive and learned olfactory preferences, and switch between those preferences, is poorly understood. Here, we map an olfactory network, from sensory input to motor output, which regulates the learned olfactory aversion of Caenorhabditis elegans for the smell of pathogenic bacteria. Naive animals prefer smells of pathogens but animals trained with pathogens lose this attraction. We find that two different neural circuits subserve these preferences, with one required for the naive preference and the other specifically for the learned preference. Calcium imaging and behavioral analysis reveal that the naive preference reflects the direct transduction of the activity of olfactory sensory neurons into motor response, whereas the learned preference involves modulations to signal transduction to downstream neurons to alter motor response. Thus, two different neural circuits regulate a behavioral switch between naive and learned olfactory preferences.Organismic and Evolutionary BiologyPhysic

Family function and eating behaviours among Hispanic/Latino youth: results from the Hispanic Community Children’s Health Study/Study of Latino Youth (SOL Youth)

Objective: To elucidate mechanisms across family function, home environment and eating behaviours within sociocultural context among Hispanic youth. Design: Two models tested via path analysis (youth fruit and vegetable (FV) con- sumption; empty energy consumption) using data from the Study of Latino Youth (2011–2013). Setting: Chicago, IL; Miami, FL; Bronx, NY; San Diego, CA. Participants: Youth (8–16-year-olds), n 1466. Results: Youth ate 2·4 servings of FV per d and received 27 % of total energy from empty energies. Perceiving higher acculturative stress was indirectly associated with lower FV consumption via a pathway of low family function and family support for FV (β = −0·013, P 12-year-olds was indirectly associated with lower FV consumption via lower family closeness and family support (β = −0·006, P < 0·001). Household food security was indirectly associated with greater FV consumption via family closeness and family support (β=0·005, P = 0·003). In contrast, perceiving higher acculturative stress was indirectly associated with higher empty energy consumption (via family closeness and family support: β = 0·003, P = 0·028 and via low family function and low family support: β = 0·008, P = 0·05). Being older was associated with higher consumption of empty energies via family closeness (related to family support: β = 0·04, P = 0·016; parenting strategies for eating: β = 0·002, P = 0·049). Conclusions: Findings suggest pathways of influence across demographic and sociocultural context, family dynamics and home environment. The directionality of these associations needs confirmation using longitudinal data.UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Sociales::Instituto de Investigaciones Psicológicas (IIP

Midline Signalling Systems Direct the Formation of a Neural Map by Dendritic Targeting in the Drosophila Motor System

During embryonic development of the motor system of Drosophila, motorneurons target their dendrites to different regions along the body axis in response to midline guidance cues

Comparative genomics of proteins involved in RNA nucleocytoplasmic export

Background: The establishment of the nuclear membrane resulted in the physical separation of transcription and translation, and presented early eukaryotes with a formidable challenge: how to shuttle RNA from the nucleus to the locus of protein synthesis. In prokaryotes, mRNA is translated as it is being synthesized, whereas in eukaryotes mRNA is synthesized and processed in the nucleus, and it is then exported to the cytoplasm. In metazoa and fungi, the different RNA species are exported from the nucleus by specialized pathways. For example, tRNA is exported by exportin-t in a RanGTP-dependent fashion. By contrast, mRNAs are associated to ribonucleoproteins (RNPs) and exported by an essential shuttling complex (TAP-p15 in human, Mex67-mtr2 in yeast) that transports them through the nuclear pore. The different RNA export pathways appear to be well conserved among members of Opisthokonta, the eukaryotic supergroup that includes Fungi and Metazoa. However, it is not known whether RNA export in the other eukaryotic supergroups follows the same export routes as in opisthokonts. Methods: Our objective was to reconstruct the evolutionary history of the different RNA export pathways across eukaryotes. To do so, we screened an array of eukaryotic genomes for the presence of homologs of the proteins involved in RNA export in Metazoa and Fungi, using human and yeast proteins as queries. Results: Our genomic comparisons indicate that the basic components of the RanGTP-dependent RNA pathways are conserved across eukaryotes, and thus we infer that these are traceable to the last eukaryotic common ancestor (LECA). On the other hand, several of the proteins involved in RanGTP-independent mRNA export pathways are less conserved, which would suggest that they represent innovations that appeared later in the evolution of eukaryotes. Conclusions: Our analyses suggest that the LECA possessed the basic components of the different RNA export mechanisms found today in opisthokonts, and that these mechanisms became more specialized throughout eukaryotic evolution

Early Release Science of the Exoplanet WASP-39b with JWST NIRSpec G395H

Measuring the abundances of carbon and oxygen in exoplanet atmospheres is considered a crucial avenue for unlocking the formation and evolution of exoplanetary systems. Access to an exoplanet's chemical inventory requires high-precision observations, often inferred from individual molecular detections with low-resolution space-based and high-resolution ground-based facilities. Here we report the medium-resolution (R$\sim$600) transmission spectrum of an exoplanet atmosphere between 3-5 $\mu$m covering multiple absorption features for the Saturn-mass exoplanet WASP-39b, obtained with JWST NIRSpec G395H. Our observations achieve 1.46x photon precision, providing an average transit depth uncertainty of 221 ppm per spectroscopic bin, and present minimal impacts from systematic effects. We detect significant absorption from CO$_2$ (28.5$\sigma$) and H$_2$O (21.5$\sigma$), and identify SO$_2$ as the source of absorption at 4.1 $\mu$m (4.8$\sigma$). Best-fit atmospheric models range between 3 and 10x solar metallicity, with sub-solar to solar C/O ratios. These results, including the detection of SO$_2$, underscore the importance of characterising the chemistry in exoplanet atmospheres, and showcase NIRSpec G395H as an excellent mode for time series observations over this critical wavelength range.Comment: 44 pages, 11 figures, 3 tables. Resubmitted after revision to Natur

The James Webb Space Telescope Mission

Twenty-six years ago a small committee report, building on earlier studies, expounded a compelling and poetic vision for the future of astronomy, calling for an infrared-optimized space telescope with an aperture of at least $4m$. With the support of their governments in the US, Europe, and Canada, 20,000 people realized that vision as the $6.5m$ James Webb Space Telescope. A generation of astronomers will celebrate their accomplishments for the life of the mission, potentially as long as 20 years, and beyond. This report and the scientific discoveries that follow are extended thank-you notes to the 20,000 team members. The telescope is working perfectly, with much better image quality than expected. In this and accompanying papers, we give a brief history, describe the observatory, outline its objectives and current observing program, and discuss the inventions and people who made it possible. We cite detailed reports on the design and the measured performance on orbit.Comment: Accepted by PASP for the special issue on The James Webb Space Telescope Overview, 29 pages, 4 figure

Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Early Release Science of the exoplanet WASP-39b with JWST NIRSpec G395H

This is the author accepted manuscript. The final version is available from Nature Research via the DOI in this recordData Availability: The data used in this paper are associated with JWST program ERS 1366 (observation #4) and are available from the Mikulski Archive for Space Telescopes (https://mast.stsci.edu). Science data processing version (SDP_VER) 2022_2a generated the uncalibrated data that we downloaded from MAST. We used JWST Calibration Pipeline software version (CAL_VER) 1.5.3 with modifications described in the text. We used calibration reference data from context (CRDS_CTX) 0916, except as noted in the text. All the data and models presented in this publication can be found at 10.5281/zenodo.7185300.Code Availability: The codes used in this publication to extract, reduce and analyze the data are as follows; STScI JWST Calibration pipeline45 (https://github.com/spacetelescope/jwst), Eureka!53 (https://eurekadocs.readthedocs.io/en/latest/), ExoTiC-JEDI47 (https://github.com/ExoTiC/ExoTiC-JEDI), juliet71 (https://juliet.readthedocs.io/en/latest/), Tiberius15,49,50, transitspectroscopy51 (https://github.com/nespinoza/transitspectroscopy). In addition, these made use of batman65 (http://lkreidberg.github.io/batman/docs/html/index.html), celerite86 (https://celerite.readthedocs.io/en/stable/), chromatic (https://zkbt.github.io/chromatic/), Dynesty72 (https://dynesty.readthedocs.io/en/stable/index.html), emcee69 (https://emcee.readthedocs.io/en/stable/), exoplanet83 (https://docs.exoplanet.codes/en/latest/), ExoTEP75–77, ExoTHETyS79 (https://github.com/ucl-exoplanets/ExoTETHyS), ExoTiCISM57 (https://github.com/Exo-TiC/ExoTiC-ISM), ExoTiC-LD58 (https://exoticld.readthedocs.io/en/latest/), george68 (https://george.readthedocs.io/en/latest/) JAX82 (https://jax.readthedocs.io/en/latest/), LMFIT70 (https://lmfit.github.io/lmfit-py/), Pylightcurve78 (https://github.com/ucl-exoplanets/pylightcurve), Pymc3138 (https://docs.pymc.io/en/v3/index.html) and Starry84 (https://starry.readthedocs.io/en/latest/), each of which use the standard python libraries astropy139,140, matplotlib141, numpy142, pandas143, scipy64 and xarray144. The atmospheric models used to fit the data can be found at ATMO[Tremblin2015,Drummond2016,Goyal2018,Goyal2020]88–91, PHOENIX92–94, PICASO98,99 (https://natashabatalha.github.io/picaso/), Virga98,107 (https://natashabatalha.github.io/virga/), and gCMCRT115 (https://github.com/ELeeAstro/gCMCRT).Measuring the abundances of carbon and oxygen in exoplanet atmospheres is considered a crucial avenue for unlocking the formation and evolution of exoplanetary systems. Access to an exoplanet’s chemical inventory requires high precision observations, often inferred from individual molecular detections with low-resolution space-based and high-resolution ground-based facilities. Here we report the medium-resolution (R≈600) transmission spectrum of an exoplanet atmosphere between 3–5 μm covering multiple absorption features for the Saturn-mass exoplanet WASP-39b, obtained with JWST NIRSpec G395H. Our observations achieve 1.46× photon precision, providing an average transit depth uncertainty of 221 ppm per spectroscopic bin, and present minimal impacts from systematic effects. We detect significant absorption from CO2 (28.5σ ) and H2O (21.5σ ), and identify SO2 as the source of absorption at 4.1 μ m (4.8σ ). Best-fit atmospheric models range between 3× and 10× solar metallicity, with sub-solar to solar C/O ratios. These results, including the detection of SO2, underscore the importance of characterising the chemistry in exoplanet atmospheres, and showcase NIRSpec G395H as an excellent mode for time series observations over this critical wavelength range.Science and Technology Facilities Council (STFC)UKR