Maintaining Muscle Mitochondria via Transsynaptic Signaling

Dominant VAPB mutations are implicated in neurodegenerative disease, including amyotrophic lateral sclerosis and spinal muscular atrophy. In the current issue, Han et al. (2012) uncover a mechanism through which the secreted VAPB MSP domain regulates actin organization and mitochondrial function in muscle cells through LAR and Robo receptor activation

Drosophila semaphorin2b is Required for the Axon Guidance of a Subset of Embryonic Neurons

Background: The process of axon guidance is important in establishing functional neural circuits. The differential expression of cell-autonomous axon guidance factors is crucial for allowing axons of different neurons to take unique trajectories in response to spatially and temporally restricted cell non-autonomous axon guidance factors. A key motivation in the field is to provide adequate explanations for axon behavior with respect to the differential expression of these factors. Results: We report the characterization of a predicted secreted semaphorin family member, semaphorin2b (Sema-2b) in Drosophila embryonic axon guidance. Misexpression of Sema-2b in neurons causes highly penetrant axon guidance phenotypes in specific longitudinal and motoneuron pathways; however, expression of Sema-2b in muscles traversed by these motoneurons has no effect on axon guidance. In Sema-2b loss-of-function embryos, specific motoneuron and interneuron axon pathways display guidance defects. Specific visualization of the neurons that normally express Sema-2b reveals that this neuronal cohort is strongly affected by Sema-2b loss-of-function alleles. Conclusions: While secreted semaphorins have been implicated as cell non-autonomous chemorepellants in a variety of contexts, here we report previously undescribed Sema-2b loss-of-function and misexpression phenotypes that are consistent with a cell-autonomous role for Sema-2b. Developmental Dynamics 242:861–873, 2013. © 2013 Wiley Periodicals, Inc. Key Findings Misexpression of the secreted semaphorin Sema-2b in neurons results in specific axon guidance phenotypes. Both Sema-2b loss-of-function and misexpression phenotypes are congruent with a cell-autonomous role for Sema-2b. Novel axon guidance phenotypes caused by Sema-2b loss-of-function mutations are characterized

VEGF-A and Semaphorin3A: Modulators of vascular sympathetic innervation

AbstractSympathetic nerve activity regulates blood pressure by altering peripheral vascular resistance. Variations in vascular sympathetic innervation suggest that vascular-derived cues promote selective innervation of particular vessels during development. As axons extend towards peripheral targets, they migrate along arterial networks following gradients of guidance cues. Collective ratios of these gradients may determine whether axons grow towards and innervate vessels or continue past non-innervated vessels towards peripheral targets. Utilizing directed neurite outgrowth in a three-dimensional (3D) co-culture, we observed increased axon growth from superior cervical ganglion explants (SCG) towards innervated compared to non-innervated vessels, mediated in part by vascular endothelial growth factor (VEGF-A) and Semaphorin3A (Sema3A) which both signal via neuropilin-1 (Nrp1). Exogenous VEGF-A, delivered by high-expressing VEGF-A–LacZ vessels or by rhVEGF-A/alginate spheres, increased sympathetic neurite outgrowth while exogenous rhSema3A/Fc decreased neurite outgrowth. VEGF-A expression is similar between the innervated and non-innervated vessels examined. Sema3A expression is higher in non-innervated vessels. Spatial gradients of Sema3A and VEGF-A may promote differential Nrp1 binding. Vessels expressing high levels of Sema3A favor Nrp1-PlexinA1 signaling, producing chemorepulsive cues limiting sympathetic neurite outgrowth and vascular innervation; while low Sema3A expressing vessels favor Nrp1-VEGFR2 signaling providing chemoattractive cues for sympathetic neurite outgrowth and vascular innervation

Single-Cell Phenotyping within Transparent Intact Tissue through Whole-Body Clearing

Understanding the structure-function relationships at cellular, circuit, and organ-wide scale requires 3D anatomical and phenotypical maps, currently unavailable for many organs across species. At the root of this knowledge gap is the absence of a method that enables whole-organ imaging. Herein, we present techniques for tissue clearing in which whole organs and bodies are rendered macromolecule-permeable and optically transparent, thereby exposing their cellular structure with intact connectivity. We describe PACT (passive clarity technique), a protocol for passive tissue clearing and immunostaining of intact organs; RIMS (refractive index matching solution), a mounting media for imaging thick tissue; and PARS (perfusion-assisted agent release in situ), a method for whole-body clearing and immunolabeling. We show that in rodents PACT, RIMS, and PARS are compatible with endogenous-fluorescence, immunohistochemistry, RNA single-molecule FISH, long-term storage, and microscopy with cellular and subcellular resolution. These methods are applicable for high-resolution, high-content mapping and phenotyping of normal and pathological elements within intact organs and bodies

Abelson Phosphorylation of CLASP2 Modulates its Association With Microtubules and Actin

The Abelson (Abl) non-receptor tyrosine kinase regulates the cytoskeleton during multiple stages of neural development, from neurulation, to the articulation of axons and dendrites, to synapse formation and maintenance. We previously showed that Abl is genetically linked to the microtubule (MT) plus end tracking protein (+TIP) CLASP in Drosophila. Here we show in vertebrate cells that Abl binds to CLASP and phosphorylates it in response to serum or PDGF stimulation. In vitro, Abl phosphorylates CLASP with a Km of 1.89 µM, indicating that CLASP is a bona fide substrate. Abl-phosphorylated tyrosine residues that we detect in CLASP by mass spectrometry lie within previously mapped F-actin and MT plus end interaction domains. Using purified proteins, we find that Abl phosphorylation modulates direct binding between purified CLASP2 with both MTs and actin. Consistent with these observations, Abl-induced phosphorylation of CLASP2 modulates its localization as well as the distribution of F-actin structures in spinal cord growth cones. Our data suggest that the functional relationship between Abl and CLASP2 is conserved and provides a means to control the CLASP2 association with the cytoskeleton. © 2014 The Authors. Cytoskeleton Published by Wiley Periodicals, Inc

Dynamical masses and stellar evolutionary model predictions of M stars

Funding: J.P. gratefully acknowledges the support of the National Science Foundation (NSF) Graduate Research Fellowship through grant Nos. DGE1144152 and DGE1745303. K.I.Ö. gratefully acknowledges the support of the Simons Foundation through a Simons Collaboration on the Origins of Life (SCOL) PI grant (No. 321183). G.J.H. is supported by general grant 11773002 awarded by the National Science Foundation of China. L.I.C. gratefully acknowledges support from the David and Lucille Packard Foundation, the Virginia Space Grant Consortium, and Johnson & Johnson’s WiSTEM2D Award. V.V.G. gratefully acknowledges support from FONDECYT Iniciación 11180904. Support for this work was also provided by NASA through the NASA Hubble Fellowship grant Nos. HST-HF2-51460.001-A, HST-HF2-51405.001-A, and HST-HF2-51429.001-A awarded by the Space Telescope Science Institute, which is operated by the Association of Universities for Research in Astronomy, Inc., for NASA, under contract NAS5-26555.In this era of Gaia and ALMA, dynamical stellar mass measurements, derived from spatially and spectrally resolved observations of the Keplerian rotation of circumstellar disks, provide benchmarks that are independent of observations of stellar characteristics and their uncertainties. These benchmarks can then be used to validate and improve stellar evolutionary models, the latter of which can lead to both imprecise and inaccurate mass predictions for pre-main-sequence, low-mass (≤0.5 M⊙) stars. We present the dynamical stellar masses derived from disks around three M stars (FP Tau, J0432+1827, and J1100-7619) using ALMA observations of 12CO (J = 2-1) and 13CO (J = 2-1) emission. These are the first dynamical stellar mass measurements for J0432+1827 and J1100-7619 (0.192 ± 0.005 M⊙ and 0.461 ± 0.057 M⊙, respectively) and the most precise measurement for FP Tau (0.395 ± 0.012 M⊙). Fiducial stellar evolutionary model tracks, which do not include any treatment of magnetic activity, agree with the dynamical stellar mass measurement of J0432+1827 but underpredict the mass by ∼60% for FP Tau and by ∼80% for J1100-7619. Possible explanations for the underpredictions include inaccurate assumptions of stellar effective temperature, undetected binarity for J1100-7619, and that fiducial stellar evolutionary models are not complex enough to represent these stars. In the former case, the stellar effective temperatures would need to be increased by amounts ranging from ∼40 to ∼340 K to reconcile the fiducial stellar evolutionary model predictions with the dynamically measured masses. In the latter case, we show that the dynamical masses can be reproduced using results from stellar evolutionary models with starspots, which incorporate fractional starspot coverage to represent the manifestation of magnetic activity. Folding in low-mass M stars from the literature and assuming that the stellar effective temperatures are imprecise but accurate, we find tentative evidence of a relationship between fractional starspot coverage and observed effective temperature for these young, cool stars.Publisher PDFPeer reviewe

DHODH modulates transcriptional elongation in the neural crest and melanoma

Melanoma is a tumour of transformed melanocytes, which are originally derived from the embryonic neural crest. It is unknown to what extent the programs that regulate neural crest development interact with mutations in the BRAF oncogene, which is the most commonly mutated gene in human melanoma1. We have used zebrafish embryos to identify the initiating transcriptional events that occur on activation of human BRAF(V600E) (which encodes an amino acid substitution mutant of BRAF) in the neural crest lineage. Zebrafish embryos that are transgenic for mitfa:BRAF(V600E) and lack p53 (also known as tp53) have a gene signature that is enriched for markers of multipotent neural crest cells, and neural crest progenitors from these embryos fail to terminally differentiate. To determine whether these early transcriptional events are important for melanoma pathogenesis, we performed a chemical genetic screen to identify small-molecule suppressors of the neural crest lineage, which were then tested for their effects on melanoma. One class of compound, inhibitors of dihydroorotate dehydrogenase (DHODH), for example leflunomide, led to an almost complete abrogation of neural crest development in zebrafish and to a reduction in the self-renewal of mammalian neural crest stem cells. Leflunomide exerts these effects by inhibiting the transcriptional elongation of genes that are required for neural crest development and melanoma growth. When used alone or in combination with a specific inhibitor of the BRAF(V600E) oncogene, DHODH inhibition led to a marked decrease in melanoma growth both in vitro and in mouse xenograft studies. Taken together, these studies highlight developmental pathways in neural crest cells that have a direct bearing on melanoma formation

HO_x chemistry during INTEX-A 2004: Observation, model calculation, and comparison with previous studies

OH and HO_2 were measured with the Airborne Tropospheric Hydrogen Oxides Sensor (ATHOS) as part of a large measurement suite from the NASA DC-8 aircraft during the Intercontinental Chemical Transport Experiment-A (INTEX-A). This mission, which was conducted mainly over North America and the western Atlantic Ocean in summer 2004, was an excellent test of atmospheric oxidation chemistry. The HOx results from INTEX-A are compared to those from previous campaigns and to results for other related measurements from INTEX-A. Throughout the troposphere, observed OH was generally 0.95 of modeled OH; below 8 km, observed HO_2 was generally 1.20 of modeled HO_2. This observed-to-modeled comparison is similar to that for TRACE-P, another midlatitude study for which the median observed-to-modeled ratio was 1.08 for OH and 1.34 for HO_2, and to that for PEM-TB, a tropical study for which the median observed-to-modeled ratio was 1.17 for OH and 0.97 for HO_2. HO_2 behavior above 8 km was markedly different. The observed-to-modeled HO_2 ratio increased from ∼1.2 at 8 km to ∼3 at 11 km with the observed-to-modeled ratio correlating with NO. Above 8 km, the observed-to-modeled HO_2 and observed NO were both considerably greater than observations from previous campaigns. In addition, the observed-to-modeled HO_2/OH, which is sensitive to cycling reactions between OH and HO_2, increased from ∼1.5 at 8 km to almost 3.5 at 11 km. These discrepancies suggest a large unknown HO_x source and additional reactants that cycle HO_x from OH to HO_2. In the continental planetary boundary layer, the observed-to-modeled OH ratio increased from 1 when isoprene was less than 0.1 ppbv to over 4 when isoprene was greater than 2 ppbv, suggesting that forests throughout the United States are emitting unknown HO_x sources. Progress in resolving these discrepancies requires a focused research activity devoted to further examination of possible unknown OH sinks and HO_x sources

Long-term biological effects of petroleum residues on fiddler crabs in salt marshes

Author Posting. © The Author(s), 2007. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Marine Pollution Bulletin 54 (2007): 955-962, doi:10.1016/j.marpolbul.2007.02.015.In September 1969,the Florida barge spilled 700,000 L of No. 2 fuel oil into the salt marsh sediments of Wild Harbor (Buzzards Bay, MA). Today the aboveground environment appears unaffected, but a substantial amount of moderately degraded petroleum still remains 8 to 20 cm below the surface. The salt marsh fiddler crabs, Uca pugnax, which burrow into the sediments at depths of 5 to 25 cm, are chronically exposed to the spilled oil. Behavioral studies conducted with U. pugnax from Wild Harbor and a control site, Great Sippewissett marsh, found that crabs exposed to the oil avoided burrowing into oiled layers, suffered delayed escape responses, lowered feeding rates, and lower densities. The oil residues are therefore biologically active and affect U. pugnax populations. Our results add new knowledge about long-term consequences of spilled oil, a dimension that should be included when assessing oil-impacted areas and developing management plans designed to restore, rehabilitate, or replace impacted areas.This work was funded by a grant from the Woods Hole Oceanographic Institution Sea Grant Program, under grants from the National Oceanic and Atmospheric Administration, U.S. Department of Commerce, under Grant No. NA16RG2273, project no. R/P-73. Additional support was provided by funding from the NSF funded Research Experience for Undergraduates program, award 0453292, an Office of Naval Research Young Investigator Award (N00014-04-01-0029) to C. Reddy, and an USEPA Science to Achieve Results Graduate Fellowship (FP91661801) to E. Peacock