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190 Publications

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    Looger LabSchreiter Lab
    01/01/12 | Neural activity imaging with genetically encoded calcium indicators.
    Tian L, Akerboom J, Schreiter ER, Looger LL
    Progress in Brain Research. 2012;196:79-94. doi: 10.1016/B978-0-444-59426-6.00005-7

    Genetically encoded calcium indicators (GECIs), together with modern microscopy, allow repeated activity measurement, in real time and with cellular resolution, of defined cellular populations. Recent efforts in protein engineering have yielded several high-quality GECIs that facilitate new applications in neuroscience. Here, we summarize recent progress in GECI design, optimization, and characterization, and provide guidelines for selecting the appropriate GECI for a given biological application. We focus on the unique challenges associated with imaging in behaving animals.

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    01/01/12 | Neuronal spike generation mechanism as an oversampling, noise-shaping A-to-D converter.
    Chklovskii DB, Soudry D
    Advances in Neural Information Processing Systems. 2012;24:503-11

    We explore the hypothesis that the neuronal spike generation mechanism is an analog-to-digital converter, which rectifies low-pass filtered summed synaptic currents and encodes them into spike trains linearly decodable in post-synaptic neurons. To digitally encode an analog current waveform, the sampling rate of the spike generation mechanism must exceed its Nyquist rate. Such oversampling is consistent with the experimental observation that the precision of the spike-generation mechanism is an order of magnitude greater than the cut-off frequency of dendritic low-pass filtering. To achieve additional reduction in the error of analog-to-digital conversion, electrical engineers rely on noise-shaping. If noise-shaping were used in neurons, it would introduce correlations in spike timing to reduce low-frequency (up to Nyquist) transmission error at the cost of high-frequency one (from Nyquist to sampling rate). Using experimental data from three different classes of neurons, we demonstrate that biological neurons utilize noise-shaping. We also argue that rectification by the spike-generation mechanism may improve energy efficiency and carry out de-noising. Finally, the zoo of ion channels in neurons may be viewed as a set of predictors, various subsets of which are activated depending on the statistics of the input current.

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    01/01/12 | Rapid mounting of adult Drosophila structures in Hoyer's medium.
    Stern DL, Sucena E
    Cold Spring Harb Protoc. 2012 Jan;2012(1):107-9. doi: 10.1101/pdb.prot067371

    The Drosophila cuticle carries a rich array of morphological details. Thus, cuticle examination has had a central role in the history of genetics. This protocol describes a procedure for mounting adult cuticles in Hoyer's medium, a useful mountant for both larval and adult cuticles. The medium digests soft tissues rapidly, leaving the cuticle cleared for observation. In addition, samples can be transferred directly from water to Hoyer's medium. However, specimens mounted in Hoyer's medium degrade over time. For example, the fine denticles on the larval dorsum are best observed soon after mounting; they begin to fade after 1 week, and can disappear completely after several months. More robust features, such as the ventral denticle belts, will persist for a longer period of time. Because adults cannot profitably be mounted whole in Hoyer's medium, some dissection is necessary.

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    Looger Lab
    01/01/12 | Running in reverse: rhodopsins sense voltage.
    Looger LL
    Nature Methods. 2012 Jan;9(1):43-4. doi: 10.1038/nmeth.1817
    Singer Lab
    01/01/12 | Spatial arrangement of an RNA zipcode identifies mRNAs under post-transcriptional control.
    Patel VL, Mitra S, Harris R, Buxbaum AR, Lionnet T, Brenowitz M, Girvin M, Levy M, Almo SC, Singer RH, Chao JA
    Genes & Development. 2012 Jan 1;26(1):43-53. doi: 10.1101/gad.177428.111

    How RNA-binding proteins recognize specific sets of target mRNAs remains poorly understood because current approaches depend primarily on sequence information. In this study, we demonstrate that specific recognition of messenger RNAs (mRNAs) by RNA-binding proteins requires the correct spatial positioning of these sequences. We characterized both the cis-acting sequence elements and the spatial restraints that define the mode of RNA binding of the zipcode-binding protein 1 (ZBP1/IMP1/IGF2BP1) to the β-actin zipcode. The third and fourth KH (hnRNP K homology) domains of ZBP1 specifically recognize a bipartite RNA element comprised of a 5' element (CGGAC) followed by a variable 3' element (C/A-CA-C/U) that must be appropriately spaced. Remarkably, the orientation of these elements is interchangeable within target transcripts bound by ZBP1. The spatial relationship of this consensus binding site identified conserved transcripts that were verified to associate with ZBP1 in vivo. The dendritic localization of one of these transcripts, spinophilin, was found to be dependent on both ZBP1 and the RNA elements recognized by ZBP1 KH34.

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    10/01/12 | Super-resolution using sparse representations over learned dictionaries: reconstruction of brain structure using electron microscopy.
    Hu T, Nunez-Iglesias J, Vitaladevuni S, Scheffer L, Xu S, Bolorizadeh M, Hess H, Fetter R, Chklovskii D . 2012 Oct:

    A central problem in neuroscience is reconstructing neuronal circuits on the synapse level. Due to a wide range of scales in brain architecture such reconstruction requires imaging that is both high-resolution and high-throughput. Existing electron microscopy (EM) techniques possess required resolution in the lateral plane and either high-throughput or high depth resolution but not both. Here, we exploit recent advances in unsupervised learning and signal processing to obtain high depth-resolution EM images computationally without sacrificing throughput. First, we show that the brain tissue can be represented as a sparse linear combination of localized basis functions that are learned using high-resolution datasets. We then develop compressive sensing-inspired techniques that can reconstruct the brain tissue from very few (typically 5) tomographic views of each section. This enables tracing of neuronal processes and, hence, high throughput reconstruction of neural circuits on the level of individual synapses.

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    01/01/12 | The Pfam protein families database.
    Punta M, Coggill PC, Eberhardt RY, Mistry J, Tate J, Boursnell C, Pang N, Forslund K, Ceric G, Clements J, Heger A, Holm L, Sonnhammer EL, Sean R. Eddy , Bateman A, Finn RD
    Nucleic acids research. 2012 Jan;40:D290-301. doi: 10.1093/nar/gkr1065

    Pfam is a widely used database of protein families, currently containing more than 13,000 manually curated protein families as of release 26.0. Pfam is available via servers in the UK (, the USA ( and Sweden ( Here, we report on changes that have occurred since our 2010 NAR paper (release 24.0). Over the last 2 years, we have generated 1840 new families and increased coverage of the UniProt Knowledgebase (UniProtKB) to nearly 80%. Notably, we have taken the step of opening up the annotation of our families to the Wikipedia community, by linking Pfam families to relevant Wikipedia pages and encouraging the Pfam and Wikipedia communities to improve and expand those pages. We continue to improve the Pfam website and add new visualizations, such as the ’sunburst’ representation of taxonomic distribution of families. In this work we additionally address two topics that will be of particular interest to the Pfam community. First, we explain the definition and use of family-specific, manually curated gathering thresholds. Second, we discuss some of the features of domains of unknown function (also known as DUFs), which constitute a rapidly growing class of families within Pfam.

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    Cardona Lab
    01/01/12 | TrakEM2 software for neural circuit reconstruction.
    Cardona A, Saalfeld S, Schindelin J, Arganda-Carreras I, Preibisch S, Longair M, Tomancak P, Hartenstein V, Douglas RJ
    PLoS One. 2012;7:e38011. doi: 10.1371/journal.pone.0038011

    A key challenge in neuroscience is the expeditious reconstruction of neuronal circuits. For model systems such as Drosophila and C. elegans, the limiting step is no longer the acquisition of imagery but the extraction of the circuit from images. For this purpose, we designed a software application, TrakEM2, that addresses the systematic reconstruction of neuronal circuits from large electron microscopical and optical image volumes. We address the challenges of image volume composition from individual, deformed images; of the reconstruction of neuronal arbors and annotation of synapses with fast manual and semi-automatic methods; and the management of large collections of both images and annotations. The output is a neural circuit of 3d arbors and synapses, encoded in NeuroML and other formats, ready for analysis.

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    01/01/12 | Transcriptional control of cell fate specification: lessons from the fly retina.
    Quan X, Ramaekers A, Hassan BA
    Current Topics in Developmental Biology. 2012;98:259-76. doi: 10.1016/B978-0-12-386499-4.00010-0

    It is now widely recognized that as cells of developing tissues transition through successive states of decreasing pluripotency into a state of terminal differentiation, they undergo significant changes in their gene expression profiles. Interestingly, these successive states of increasing differentiation are marked by the spatially and temporally restricted expression of sets of transcription factors. Each wave of transcription factors not only signals the arrival of a given stage in cellular differentiation, but it is also necessary for the activation of the next set of transcription factors, creating the appearance of a smooth, directed, and deterministic genetic program of cellular differentiation. Until recently, however, it was largely unknown which genes, besides each other, these transcription factors were activating. Thus, the molecular definition of any given step of differentiation, and how it gave rise to the following step remained unclear. Recent advances in transcriptomics, bioinformatics, and molecular genetics resulted in the identification of numerous transcription factor target genes (TGs). These advances have opened the door to using similar approaches in developmental biology to understand what the transcriptional cascades of cellular differentiation might be. Using the development of the Drosophila eye as a model system, we discuss the role of transcription factors and their TGs in cell fate specification and terminal differentiation.

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    01/01/12 | Use of a Drosophila genome-wide conserved sequence database to identify functionally related cis-regulatory enhancers.
    Brody T, Yavatkar AS, Kuzin A, Kundu M, Tyson LJ, Ross J, Lin T, Lee C, Awasaki T, Lee T, Odenwald WF
    Developmental Dynamics: An Official Publication of the American Association of Anatomists. 2012 Jan;241:169-89. doi: 10.1002/dvdy.22728

    Phylogenetic footprinting has revealed that cis-regulatory enhancers consist of conserved DNA sequence clusters (CSCs). Currently, there is no systematic approach for enhancer discovery and analysis that takes full-advantage of the sequence information within enhancer CSCs.

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