A mammalian enhancer trap resource for discovering and manipulating neuronal cell types

  1. Yasuyuki Shima
  2. Ken Sugino
  3. Chris Hempel
  4. Masami Shima
  5. Praveen Taneja
  6. James B Bullis
  7. Sonam Mehta
  8. Carlos Lois
  9. Sacha B Nelson  Is a corresponding author
  1. Brandeis University, United States
  2. Janelia Research Campus, Howard Hughes Medical Institute, United States
  3. Galenea Corporation, United States
  4. California Institute of Technology, United States

Abstract

There is a continuing need for driver strains to enable cell type-specific manipulation in the nervous system. Each cell type expresses a unique set of genes, and recapitulating expression of marker genes by BAC transgenesis or knock-in has generated useful transgenic mouse lines. However since genes are often expressed in many cell types, many of these lines have relatively broad expression patterns. We report an alternative transgenic approach capturing distal enhancers for more focused expression. We identified an enhancer trap probe often producing restricted reporter expression and developed efficient enhancer trap screening with the PiggyBac transposon. We established more than 200 lines and found many lines that label small subsets of neurons in brain substructures, including known and novel cell types. Images and other information about each line are available online (enhancertrap.bio.brandeis.edu).

Article and author information

Author details

  1. Yasuyuki Shima

    Department of Biology and Center for Behavioral Genomics, Brandeis University, Waltham, United States
    Competing interests
    No competing interests declared.
  2. Ken Sugino

    Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States
    Competing interests
    No competing interests declared.
  3. Chris Hempel

    Galenea Corporation, Wakefield, United States
    Competing interests
    No competing interests declared.
  4. Masami Shima

    Department of Biology and National Center for Behavioral Genomics, Brandeis University, Waltham, United States
    Competing interests
    No competing interests declared.
  5. Praveen Taneja

    Department of Biology and National Center for Behavioral Genomics, Brandeis University, Waltham, United States
    Competing interests
    No competing interests declared.
  6. James B Bullis

    Department of Biology and National Center for Behavioral Genomics, Brandeis University, Waltham, United States
    Competing interests
    No competing interests declared.
  7. Sonam Mehta

    Department of Biology and National Center for Behavioral Genomics, Brandeis University, Waltham, United States
    Competing interests
    No competing interests declared.
  8. Carlos Lois

    Division of Biology and Biological Engineering Beckman Institute, California Institute of Technology, Pasadena, United States
    Competing interests
    No competing interests declared.
  9. Sacha B Nelson

    Department of Biology and National Center for Behavioral Genomics, Brandeis University, Waltham, United States
    For correspondence
    nelson@brandeis.edu
    Competing interests
    Sacha B Nelson, Reviewing editor, eLife.

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols (#14004) of Brandeis University. All surgery was performed under ketamine and xylazine anesthesia, and every effort was made to minimize suffering.

Copyright

© 2016, Shima et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 5,406
    views
  • 1,176
    downloads
  • 58
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Yasuyuki Shima
  2. Ken Sugino
  3. Chris Hempel
  4. Masami Shima
  5. Praveen Taneja
  6. James B Bullis
  7. Sonam Mehta
  8. Carlos Lois
  9. Sacha B Nelson
(2016)
A mammalian enhancer trap resource for discovering and manipulating neuronal cell types
eLife 5:e13503.
https://doi.org/10.7554/eLife.13503

Share this article

https://doi.org/10.7554/eLife.13503

Further reading

    1. Neuroscience
    Xiaoqian Yan, Sarah Shi Tung ... Kalanit Grill-Spector
    Research Article

    Organizing the continuous stream of visual input into categories like places or faces is important for everyday function and social interactions. However, it is unknown when neural representations of these and other visual categories emerge. Here, we used steady-state evoked potential electroencephalography to measure cortical responses in infants at 3–4 months, 4–6 months, 6–8 months, and 12–15 months, when they viewed controlled, gray-level images of faces, limbs, corridors, characters, and cars. We found that distinct responses to these categories emerge at different ages. Reliable brain responses to faces emerge first, at 4–6 months, followed by limbs and places around 6–8 months. Between 6 and 15 months response patterns become more distinct, such that a classifier can decode what an infant is looking at from their brain responses. These findings have important implications for assessing typical and atypical cortical development as they not only suggest that category representations are learned, but also that representations of categories that may have innate substrates emerge at different times during infancy.

    1. Neuroscience
    2. Stem Cells and Regenerative Medicine
    Desiree Böck, Maria Wilhelm ... Gerald Schwank
    Research Article

    Parkinson’s disease (PD) is a multifactorial disease caused by irreversible progressive loss of dopaminergic neurons (DANs). Recent studies have reported the successful conversion of astrocytes into DANs by repressing polypyrimidine tract binding protein 1 (PTBP1), which led to the rescue of motor symptoms in a chemically-induced mouse model of PD. However, follow-up studies have questioned the validity of this astrocyte-to-DAN conversion model. Here, we devised an adenine base editing strategy to downregulate PTBP1 in astrocytes and neurons in a chemically-induced PD mouse model. While PTBP1 downregulation in astrocytes had no effect, PTBP1 downregulation in neurons of the striatum resulted in the expression of the DAN marker tyrosine hydroxylase (TH) in non-dividing neurons, which was associated with an increase in striatal dopamine concentrations and a rescue of forelimb akinesia and spontaneous rotations. Phenotypic analysis using multiplexed iterative immunofluorescence imaging further revealed that most of these TH-positive cells co-expressed the dopaminergic marker DAT and the pan-neuronal marker NEUN, with the majority of these triple-positive cells being classified as mature GABAergic neurons. Additional research is needed to fully elucidate the molecular mechanisms underlying the expression of the observed markers and understand how the formation of these cells contributes to the rescue of spontaneous motor behaviors. Nevertheless, our findings support a model where downregulation of neuronal, but not astrocytic, PTBP1 can mitigate symptoms in PD mice.