SummaryDrosophila melanogaster has a rich repertoire of innate and learned behaviors. Its 100,000-neuron brain is a large but tractable target for comprehensive neural circuit mapping. Only electron microscopy (EM) enables complete, unbiased mapping of synaptic connectivity; however, the fly brain is too large for conventional EM. We developed a custom high-throughput EM platform and imaged the entire brain of an adult female fly at synaptic resolution. To validate the dataset, we traced brain-spanning circuitry involving the mushroom body (MB), which has been extensively studied for its role in learning. All inputs to Kenyon cells (KCs), the intrinsic neurons of the MB, were mapped, revealing a previously unknown cell type, postsynaptic partners of KC dendrites, and unexpected clustering of olfactory projection neurons. These reconstructions show that this freely available EM volume supports mapping of brain-spanning circuits, which will significantly accelerate Drosophila neuroscience.Video Abstract
Complex and interconnected signaling networks allow organisms to control cell division, growth, differentiation, or programmed cell death in response to metabolic and environmental cues. In plants, it is known that sugar and nitrogen are critical nutrient signals; however, our understanding of the molecular mechanisms underlying nutrient signal transduction is very limited. To begin unraveling complex sugar signaling networks in plants, DNA microarray analysis was used to determine the effects of glucose and inorganic nitrogen source on gene expression on a global scale in Arabidopsis thaliana. In whole seedling tissue, glucose is a more potent signal in regulating transcription than inorganic nitrogen. In fact, other than genes associated with nitrate assimilation, glucose had a greater effect in regulating nitrogen metabolic genes than nitrogen itself. Glucose also regulated a broader range of genes, including genes associated with carbohydrate metabolism, signal transduction, and metabolite transport. In addition, a large number of stress responsive genes were also induced by glucose, indicating a role of sugar in environmental responses. Cluster analysis revealed significant interaction between glucose and nitrogen in regulating gene expression because glucose can modulate the effects of nitrogen and vise versa. Intriguingly, cycloheximide treatment appeared to disrupt glucose induction more than glucose repression, suggesting that de novo protein synthesis is an intermediary event required before most glucose induction can occur. Cross talk between sugar and ethylene signaling may take place on the transcriptional level because several ethylene biosynthetic and signal transduction genes are repressed by glucose, and the repression is largely unaffected by cycloheximide. Collectively, our global expression data strongly support the idea that glucose and inorganic nitrogen act as both metabolites and signaling molecules.
We present results on the electroexcitation of the low mass resonances (1232)P 33 , N (1440)P 11 , N (1520)D 13 , and N (1535)S 11 in a wide range of Q 2 . The results were obtained in the comprehensive analysis of data from the Continuous Electron Beam Accelerator Facility (CEBAF) large acceptance spectrometer (CLAS) detector at the Thomas Jefferson National Accelerator Facility (JLab) on differential cross sections, longitudinally polarized beam asymmetries, and longitudinal target and beam-target asymmetries for π electroproduction off the proton. The data were analyzed using two conceptually different approaches-fixed-t dispersion relations and a unitary isobar model-allowing us to draw conclusions on the model sensitivity of the obtained electrocoupling amplitudes. The amplitudes for the (1232)P 33 show the importance of a meson-cloud contribution to quantitatively explain the magnetic dipole strength, as well as the electric and scalar quadrupole transitions. They do not show any tendency of approaching the pQCD regime for Q 2 6 GeV 2 . For the Roper resonance, N (1440)P 11 , the data provide strong evidence that this state is a predominantly radial excitation of a three-quark (3q) ground state. Measured in pion electroproduction, the transverse helicity amplitude for the N (1535)S 11 allowed us to obtain the branching ratios of this state to the πN and ηN channels via comparison with the results extracted from η electroproduction. The extensive CLAS data also enabled the extraction of the γ * p → N (1520)D 13 and N (1535)S 11 longitudinal helicity amplitudes with good precision. For the N (1535)S 11 , these results became a challenge for quark models and may be indicative of large meson-cloud contributions or of representations of this state that differ from a 3q excitation. The transverse amplitudes for the N (1520)D 13 clearly show the rapid changeover from helicity-3/2 dominance at the real photon point to helicity-1/2 dominance at Q 2 > 1 GeV 2 , confirming a long-standing prediction of the constituent quark model.
The atomic nucleus is composed of two different kinds of fermions: protons and neutrons. If the protons and neutrons did not interact, the Pauli exclusion principle would force the majority of fermions (usually neutrons) to have a higher average momentum. Our high-energy electron-scattering measurements using (12)C, (27)Al, (56)Fe, and (208)Pb targets show that even in heavy, neutron-rich nuclei, short-range interactions between the fermions form correlated high-momentum neutron-proton pairs. Thus, in neutron-rich nuclei, protons have a greater probability than neutrons to have momentum greater than the Fermi momentum. This finding has implications ranging from nuclear few-body systems to neutron stars and may also be observable experimentally in two-spin-state, ultracold atomic gas systems.
2 SUMMARY (150 words) 21Drosophila melanogaster has a rich repertoire of innate and learned behaviors. Its 100,000-22 neuron brain is a large but tractable target for comprehensive neural circuit mapping. Only 23 electron microscopy (EM) enables complete, unbiased mapping of synaptic connectivity; 24 however, the fly brain is too large for conventional EM. We developed a custom high-throughput 25 EM platform and imaged the entire brain of an adult female fly. We validated the dataset by 26 tracing brain-spanning circuitry involving the mushroom body (MB), intensively studied for its 27 role in learning. Here we describe the complete set of olfactory inputs to the MB; find a new cell 28 type providing driving input to Kenyon cells (the intrinsic MB neurons); identify neurons 29 postsynaptic to Kenyon cell dendrites; and find that axonal arbors providing input to the MB 30 calyx are more tightly clustered than previously indicated by light-level data. This freely available 31 EM dataset will significantly accelerate Drosophila neuroscience. 32 33 KEYWORDS 34Electron microscopy, connectomics, neural circuits, Drosophila melanogaster, mushroom body, 35 olfaction, image stitching 36 37 HIGHLIGHTS 38 -A complete adult fruit fly brain was imaged, using electron microscopy (EM) 39 -The EM volume enables brain-spanning mapping of neuronal circuits at the synaptic level 40 -Olfactory projection neurons cluster more tightly in mushroom body calyx than expected from 41 light-level data 42
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