Single-neuron axonal reconstruction: The search for a wiring diagram of the brain

Reconstruction of the axonal projection patterns of single neurons has been an important tool for understanding both the diversity of cell types in the brain and the logic of information flow between brain regions. Innovative approaches now enable the complete reconstruction of axonal projection patterns of individual neurons with vastly increased throughput. Here, we review how advances in genetic, imaging, and computational techniques have been exploited for axonal reconstruction. We also discuss how new innovations could enable the integration of genetic and physiological information with axonal morphology for producing a census of cell types in the mammalian brain at scale.Reconstruction of the axonal projection patterns of single neurons has been an important tool for understanding both the diversity of cell types in the brain and the logic of information flow between brain regions. Innovative approaches now enable the complete reconstruction of axonal projection patterns of individual neurons with vastly increased throughput. Here, we review how advances in genetic, imaging, and computational techniques have been exploited for axonal reconstruction. We also discuss how new innovations could enable the integration of genetic and physiological information with axonal morphology for producing a census of cell types in the mammalian brain at scale.First author draf

Long distance projections of cortical pyramidal neurons

The neuronal circuits defined by the axonal projections of pyramidal neurons in the cerebral cortex are responsible for processing sensory and other information to plan and execute behavior. Subtypes of cortical pyramidal neurons are organized across layers, with those in different layers distinguished by their patterns of axonal projections and connectivity. For example, those in layers 2 and 3 project between cortical areas to integrate sensory and other information with motor areas; while those in layers 5 and 6 also integrate information between cortical areas, but also project to subcortical structures involved in the generation of behavior. Recent advances in neuroanatomical techniques allow one to target specific subtypes of cortical pyramidal neurons and label both their inputs and projections. Combining these methods with neurophysiological recording techniques and newly introduced atlases of the mouse brain provide the opportunity to achieve a detailed view of the organization of cerebral cortical circuits. © 2016 Wiley Periodicals, Inc.This review was supported by the National Institute of Mental Health NIMH IRP (MH002497-25) (to CRG); and the Howard Hughes Medical Institute (to MNE and JC). Abbreviations: BAC bacterial artificial chromosomes; IT, inter-telencephalic neurons; PT pyramidal tract neurons; MOs, secondary motor cortex. (MH002497-25 - National Institute of Mental Health NIMH IRP; Howard Hughes Medical Institute)Accepted manuscrip

The Taste of Carbonation

Carbonated beverages are commonly available and immensely popular, but little is known about the cellular and molecular mechanisms underlying the perception of carbonation in the mouth. In mammals, carbonation elicits both somatosensory and chemosensory responses, including activation of taste neurons. We have identified the cellular and molecular substrates for the taste of carbonation. By targeted genetic ablation and the silencing of synapses in defined populations of taste receptor cells, we demonstrated that the sour-sensing cells act as the taste sensors for carbonation, and showed that carbonic anhydrase 4, a glycosylphosphatidylinositol-anchored enzyme, functions as the principal CO_2 taste sensor. Together, these studies reveal the basis of the taste of carbonation as well as the contribution of taste cells in the orosensory response to CO_2

A platform for brain-wide imaging and reconstruction of individual neurons

The structure of axonal arbors controls how signals from individual neurons are routed within the mammalian brain. However, the arbors of very few long-range projection neurons have been reconstructed in their entirety, as axons with diameters as small as 100 nm arborize in target regions dispersed over many millimeters of tissue. We introduce a platform for high-resolution, three-dimensional fluorescence imaging of complete tissue volumes that enables the visualization and reconstruction of long-range axonal arbors. This platform relies on a high-speed two-photon microscope integrated with a tissue vibratome and a suite of computational tools for large-scale image data. We demonstrate the power of this approach by reconstructing the axonal arbors of multiple neurons in the motor cortex across a single mouse brain.Howard Hughes Medical InstitutePublished versio

T2Rs Function as Bitter Taste Receptors

AbstractBitter taste perception provides animals with critical protection against ingestion of poisonous compounds. In the accompanying paper, we report the characterization of a large family of putative mammalian taste receptors (T2Rs). Here we use a heterologous expression system to show that specific T2Rs function as bitter taste receptors. A mouse T2R (mT2R-5) responds to the bitter tastant cycloheximide, and a human and a mouse receptor (hT2R-4 and mT2R-8) responded to denatonium and 6-n-propyl-2-thiouracil. Mice strains deficient in their ability to detect cycloheximide have amino acid substitutions in the mT2R-5 gene; these changes render the receptor significantly less responsive to cycloheximide. We also expressed mT2R-5 in insect cells and demonstrate specific tastant-dependent activation of gustducin, a G protein implicated in bitter signaling. Since a single taste receptor cell expresses a large repertoire of T2Rs, these findings provide a plausible explanation for the uniform bitter taste that is evoked by many structurally unrelated toxic compounds

Whole-brain profiling of cells and circuits in mammals by tissue clearing and light-sheet microscopy

Tissue clearing and light-sheet microscopy have a 100-year-plus history, yet these fields have been combined only recently to facilitate novel experiments and measurements in neuroscience. Since tissue-clearing methods were first combined with modernized light-sheet microscopy a decade ago, the performance of both technologies has rapidly improved, broadening their applications. Here, we review the state of the art of tissue-clearing methods and light-sheet microscopy and discuss applications of these techniques in profiling cells and circuits in mice. We examine outstanding challenges and future opportunities for expanding these techniques to achieve brain-wide profiling of cells and circuits in primates and humans. Such integration will help provide a systems-level understanding of the physiology and pathology of our central nervous system.P 28338 - Austrian Science Fund FWF; U01 MH105971 - NIMH NIH HHS; U01 MH114824 - NIMH NIH HHS; Howard Hughes Medical InstituteAccepted manuscrip

MouseLight Neuron AA0002

<div>A vast neural tracing effort by a team of Janelia scientists has upped the number of fully-traced neurons in the mouse brain by a factor of 10. Researchers can now download and browse the data in three dimensions. </div><div><br></div><div>Inside the mouse brain, individual neurons zigzag across hemispheres, embroider branching patterns and, researchers have now discovered, can even spool out spindly fibers up to 45 centimeters long.</div><div><br></div><div>Scientists can see and explore these wandering neural traces in 3-D in the most detailed map of mouse brain wiring yet created. The map reconstructs the shape and position of more than 300 of the 70 million neurons in the mouse brain. Previous efforts to trace the path of individual neurons had topped out in the dozens. </div><div><br></div><div>The selectively-labeled neurons were mapped in an iterative process with two-photon microscopy. The brain is sliced in 200-micron sections and a few dozen neurons are labeled at a time and imaged. Each brain imaged yields about 50 terabytes of data, each containing mapped neurons which can be browsed via the NeuronBrowser application. Researchers interested in the complete raw data set should contact Janelia to discuss obtaining it via hardware transfer. </div><div><br></div

MouseLight Neuron AA0009

<div><div>A vast neural tracing effort by a team of Janelia scientists has upped the number of fully-traced neurons in the mouse brain by a factor of 10. Researchers can now download and browse the data in three dimensions. </div><div><br></div><div>Inside the mouse brain, individual neurons zigzag across hemispheres, embroider branching patterns and, researchers have now discovered, can even spool out spindly fibers up to 45 centimeters long.</div><div><br></div><div>Scientists can see and explore these wandering neural traces in 3-D in the most detailed map of mouse brain wiring yet created. The map reconstructs the shape and position of more than 300 of the 70 million neurons in the mouse brain. Previous efforts to trace the path of individual neurons had topped out in the dozens. </div><div><br></div><div>The selectively-labeled neurons were mapped in an iterative process with two-photon microscopy. The brain is sliced in 200-micron sections and a few dozen neurons are labeled at a time and imaged. Each brain imaged yields about 50 terabytes of data, each containing mapped neurons which can be browsed via the NeuronBrowser application. Researchers interested in the complete raw data set should contact Janelia to discuss obtaining it via hardware transfer. </div></div