Microbial light-activatable proton pumps as neuronal inhibitors to functionally dissect neuronal networks in C. elegans

Essentially any behavior in simple and complex animals depends on neuronal network function. Currently, the best-defined system to study neuronal circuits is the nematode Caenorhabditis elegans, as the connectivity of its 302 neurons is exactly known. Individual neurons can be activated by photostimulation of Channelrhodopsin-2 (ChR2) using blue light, allowing to directly probe the importance of a particular neuron for the respective behavioral output of the network under study. In analogy, other excitable cells can be inhibited by expressing Halorhodopsin from Natronomonas pharaonis (NpHR) and subsequent illumination with yellow light. However, inhibiting C. elegans neurons using NpHR is difficult. Recently, proton pumps from various sources were established as valuable alternative hyperpolarizers. Here we show that archaerhodopsin-3 (Arch) from Halorubrum sodomense and a proton pump from the fungus Leptosphaeria maculans (Mac) can be utilized to effectively inhibit excitable cells in C. elegans. Arch is the most powerful hyperpolarizer when illuminated with yellow or green light while the action spectrum of Mac is more blue-shifted, as analyzed by light-evoked behaviors and electrophysiology. This allows these tools to be combined in various ways with ChR2 to analyze different subsets of neurons within a circuit. We exemplify this by means of the polymodal aversive sensory ASH neurons, and the downstream command interneurons to which ASH neurons signal to trigger a reversal followed by a directional turn. Photostimulating ASH and subsequently inhibiting command interneurons using two-color illumination of different body segments, allows investigating temporal aspects of signaling downstream of ASH

Avant-propos

L’événement de la fin de l’année académique 2002 a été le départ à la retraite de Jacques Coenen-Huther, lui qui depuis plus de vingt ans a apporté au Département de Sociologie et à l’Université de Genève sa vaste culture, sa rigueur intellectuelle, son imagination créatrice, ainsi que son enthousiasme pédagogique sans concession – ce qu’atteste une fois de plus sa contribution au présent ouvrage. Comment, à l’occasion de cette transition si décisive, manifester à Jacques notre estime, notre ..

Avant-propos

L’événement de la fin de l’année académique 2002 a été le départ à la retraite de Jacques Coenen-Huther, lui qui depuis plus de vingt ans a apporté au Département de Sociologie et à l’Université de Genève sa vaste culture, sa rigueur intellectuelle, son imagination créatrice, ainsi que son enthousiasme pédagogique sans concession – ce qu’atteste une fois de plus sa contribution au présent ouvrage. Comment, à l’occasion de cette transition si décisive, manifester à Jacques notre estime, notre ..

Optogenetic Long-Term Manipulation of Behavior and Animal Development

Channelrhodopsin-2 (ChR2) is widely used for rapid photodepolarization of neurons, yet, as it requires high-intensity blue light for activation, it is not suited for long-term in vivo applications, e.g. for manipulations of behavior, or photoactivation of neurons during development. We used “slow” ChR2 variants with mutations in the C128 residue, that exhibit delayed off-kinetics and increased light sensitivity in Caenorhabditis elegans. Following a 1 s light pulse, we could photodepolarize neurons and muscles for minutes (and with repeated brief stimulation, up to days) with low-intensity light. Photoactivation of ChR2(C128S) in command interneurons elicited long-lasting alterations in locomotion. Finally, we could optically induce profound changes in animal development: Long-term photoactivation of ASJ neurons, which regulate larval growth, bypassed the constitutive entry into the “dauer” larval state in daf-11 mutants. These lack a guanylyl cyclase, which possibly renders ASJ neurons hyperpolarized. Furthermore, photostimulated ASJ neurons could acutely trigger dauer-exit. Thus, slow ChR2s can be employed to long-term photoactivate behavior and to trigger alternative animal development

In-depth transcriptomic analysis of human retina reveals molecular mechanisms underlying diabetic retinopathy

Diabetic Retinopathy (DR) is among the major global causes for vision loss. With the rise in diabetes prevalence, an increase in DR incidence is expected. Current understanding of both the molecular etiology and pathways involved in the initiation and progression of DR is limited. Via RNA-Sequencing, we analyzed mRNA and miRNA expression profiles of 80 human post-mortem retinal samples from 43 patients diagnosed with various stages of DR. We found differentially expressed transcripts to be predominantly associated with late stage DR and pathways such as hippo and gap junction signaling. A multivariate regression model identified transcripts with progressive changes throughout disease stages, which in turn displayed significant overlap with sphingolipid and cGMP-PKG signaling. Combined analysis of miRNA and mRNA expression further uncovered disease-relevant miRNA/mRNA associations as potential mechanisms of post-transcriptional regulation. Finally, integrating human retinal single cell RNA-Sequencing data revealed a continuous loss of retinal ganglion cells, and Müller cell mediated changes in histidine and β-alanine signaling. While previously considered primarily a vascular disease, attention in DR has shifted to additional mechanisms and cell-types. Our findings offer an unprecedented and unbiased insight into molecular pathways and cell-specific changes in the development of DR, and provide potential avenues for future therapeutic intervention

Attentional distribution and spatial language

Kluth T, Schultheis H. Attentional distribution and spatial language. In: Freksa C, Nebel B, Hegarty M, Barkowsky T, eds. Spatial Cognition IX. Lecture Notes in Computer Science. Vol 8684. Springer International Publishing; 2014: 76-91.Whether visual spatial attention can be split to several discontinuous locations concurrently is still an open and intensely debated question. We address this question in the domain of spatial language use by comparing two existing and three newly proposed computational models. All models are assessed regarding their ability to account for human acceptability ratings for how well a given spatial term describes the spatial arrangement of two functionally related objects. One of the existing models assumes that taking the functional relations into account involves split attention. All new models incorporate functional relations without assuming split attention. Our simulations suggest that not assuming split attention is more appropriate for taking the functional relations into account than assuming split attention. At the same time, the simulations raise doubt as to whether any of the models appropriately captures the impact of functional relations on spatial language use

The Milky Way's bulge star formation history as constrained from its bimodal chemical abundance distribution

We conduct a quantitative analysis of the star formation history (SFH) of the Milky Way's bulge by exploiting the constraining power of its stellar [Fe/H] and [Mg/Fe] distribution functions. Using APOGEE data, we confirm the previously-established bimodal [Mg/Fe]--[Fe/H] distribution within 3 kpc of the inner Galaxy. Compared to that in the solar vicinity, the high-$\alpha$ population in the bulge peaks at a lower [Fe/H]. To fit these observations, we use a simple but flexible star formation framework, which assumes two distinct stages of gas accretion and star formation, and systematically evaluate a wide multi-dimensional parameter space. We find that the data favor a three-phase SFH that consists of an initial starburst, followed by a rapid star formation quenching episode and a lengthy, quiescent secular evolution phase. The metal-poor, high-$\alpha$ bulge stars ([Fe/H]0.15) are formed rapidly (<2 Gyr) during the early starburst. The density gap between the high- and low-$\alpha$ sequences is due to the quenching process. The metal-rich, low-$\alpha$ population ([Fe/H]>0.0 and [Mg/Fe]<0.15) then accumulates gradually through inefficient star formation during the secular phase. This is qualitatively consistent with the early SFH of the inner disk. Given this scenario, a notable fraction of young stars (age<5 Gyr) is expected to persist in the bulge. Combined with extragalactic observations, these results suggest that a rapid star formation quenching process is responsible for bimodal distributions in both the Milky Way's stellar populations and in the general galaxy population and thus plays a critical role in galaxy evolution.Comment: 16 pages, 12 figures. MNRAS in pres

The Milky Way bar and bulge revealed by APOGEE and Gaia EDR3

We investigate the inner regions of the Milky Way using data from APOGEE and Gaia EDR3. Our inner Galactic sample has more than 26 500 stars within |XGal|< 5 kpc, |YGal|< 3.5 kpc, |ZGal|< 1 kpc, and we also carry out the analysis for a foreground-cleaned subsample of 8000 stars that is more representative of the bulge-bar populations. These samples allow us to build chemo-dynamical maps of the stellar populations with vastly improved detail. The inner Galaxy shows an apparent chemical bimodality in key abundance ratios [α/Fe], [C/N], and [Mn/O], which probe different enrichment timescales, suggesting a star formation gap (quenching) between the high- and low-α populations. Using a joint analysis of the distributions of kinematics, metallicities, mean orbital radius, and chemical abundances, we can characterize the different populations coexisting in the innermost regions of the Galaxy for the first time. The chemo-kinematic data dissected on an eccentricity-|Z|max plane reveal the chemical and kinematic signatures of the bar, the thin inner disc, and an inner thick disc, and a broad metallicity population with large velocity dispersion indicative of a pressure-supported component. The interplay between these different populations is mapped onto the different metallicity distributions seen in the eccentricity-|Z|max diagram consistently with the mean orbital radius and Vφ distributions. A clear metallicity gradient as a function of |Z|max is also found, which is consistent with the spatial overlapping of different populations. Additionally, we find and chemically and kinematically characterize a group of counter-rotating stars that could be the result of a gas-rich merger event or just the result of clumpy star formation during the earliest phases of the early disc that migrated into the bulge. Finally, based on 6D information, we assign stars a probability value of being on a bar orbit and find that most of the stars with large bar orbit probabilities come from the innermost 3 kpc, with a broad dispersion of metallicity. Even stars with a high probability of belonging to the bar show chemical bimodality in the [α/Fe] versus [Fe/H] diagram. This suggests bar trapping to be an efficient mechanism, explaining why stars on bar orbits do not show a significant, distinct chemical abundance ratio signature

Two groups of red giants with distinct chemical abundances in the bulge globular cluster NGC 6553 through the eyes of APOGEE

Multiple populations revealed in globular clusters (GCs) are important windows to the formation and evolution of these stellar systems. The metal-rich GCs in the Galactic bulge are an indispensable part of this picture, but the high optical extinction in this region has prevented extensive research. In this work, we use the high-resolution near-infrared (NIR) spectroscopic data from Apache Point Observatory Galactic Evolution Experiment (APOGEE) to study the chemical abundances of NGC 6553, which is one of the most metal-rich bulge GCs. We identify 10 red giants as cluster members using their positions, radial velocities, iron abundances, and NIR photometry. Our sample stars show a mean radial velocity of −0.14 ± 5.47 km s−1, and a mean [Fe/H] of −0.15 ± 0.05. We clearly separate two populations of stars in C and N in this GC for the first time. NGC 6553 is the most metal-rich GC where the multiple stellar population phenomenon is found until now. Substantial chemical variations are also found in Na, O, and Al. However, the two populations show similar Si, Ca, and iron-peak element abundances. Therefore, we infer that the CNO, NeNa, and MgAl cycles have been activated, but the MgAl cycle is too weak to show its effect on Mg. Type Ia and Type II supernovae do not seem to have significantly polluted the second generation stars. Comparing with other GC studies, NGC 6553 shows similar chemical variations as other relatively metal-rich GCs. We also confront current GC formation theories with our results, and suggest possible avenues for improvement in the models

Desmin Knock-Out Cardiomyopathy: A Heart on the Verge of Metabolic Crisis

Desmin mutations cause familial and sporadic cardiomyopathies. In addition to perturbing the contractile apparatus, both desmin deficiency and mutated desmin negatively impact mitochondria. Impaired myocardial metabolism secondary to mitochondrial defects could conceivably exacerbate cardiac contractile dysfunction. We performed metabolic myocardial phenotyping in left ventricular cardiac muscle tissue in desmin knock-out mice. Our analyses revealed decreased mitochondrial number, ultrastructural mitochondrial defects, and impaired mitochondria-related metabolic pathways including fatty acid transport, activation, and catabolism. Glucose transporter 1 and hexokinase-1 expression and hexokinase activity were increased. While mitochondrial creatine kinase expression was reduced, fetal creatine kinase expression was increased. Proteomic analysis revealed reduced expression of proteins involved in electron transport mainly of complexes I and II, oxidative phosphorylation, citrate cycle, beta-oxidation including auxiliary pathways, amino acid catabolism, and redox reactions and oxidative stress. Thus, desmin deficiency elicits a secondary cardiac mitochondriopathy with severely impaired oxidative phosphorylation and fatty and amino acid metabolism. Increased glucose utilization and fetal creatine kinase upregulation likely portray attempts to maintain myocardial energy supply. It may be prudent to avoid medications worsening mitochondrial function and other metabolic stressors. Therapeutic interventions for mitochondriopathies might also improve the metabolic condition in desmin deficient hearts