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Elife. 2016 Sep 23;5. pii: e13918. doi: 10.7554/eLife.13918.

A robust activity marking system for exploring active neuronal ensembles.

Author information

  • 1McGovern Institute for Brain Research, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, United States.
  • 2Institute for Behavioral Genetics, University of Colorado Boulder, Boulder, United States.
  • 3Department of Genetics, University of Wisconsin-Madison, Madison, United States.
  • 4Department of Neurology, University of Wisconsin-Madison, Madison, United States.
  • 5Neuroscience Training Program, University of Wisconsin-Madison, Madison, United States.
  • 6Wellcome Trust Sanger Institute, Hinxton, United Kingdom.
  • 7Department of Psychology and Neuroscience, University of Colorado Boulder, Boulder, United States.


Understanding how the brain captures transient experience and converts it into long lasting changes in neural circuits requires the identification and investigation of the specific ensembles of neurons that are responsible for the encoding of each experience. We have developed a Robust Activity Marking (RAM) system that allows for the identification and interrogation of ensembles of neurons. The RAM system provides unprecedented high sensitivity and selectivity through the use of an optimized synthetic activity-regulated promoter that is strongly induced by neuronal activity and a modified Tet-Off system that achieves improved temporal control. Due to its compact design, RAM can be packaged into a single adeno-associated virus (AAV), providing great versatility and ease of use, including application to mice, rats, flies, and potentially many other species. Cre-dependent RAM, CRAM, allows for the study of active ensembles of a specific cell type and anatomical connectivity, further expanding the RAM system's versatility.


D. melanogaster; Npas4; RAM; Tet-Off system; activity mapping; cFos; mouse; neural ensemble; neuroscience; rat

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