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Original Research ARTICLE Provisionally accepted The full-text will be published soon. Notify me

Front. Neurosci. | doi: 10.3389/fnins.2019.00766

Monitoring behaviorally induced biochemical changes using fluorescence lifetime photometry

 Bernardo Sabatini1, 2*, Suk Joon Lee1, 2,  Yao Chen1, 2 and  Bart Lodder1, 2, 3
  • 1Harvard Medical School, United States
  • 2Howard Hughes Medical Institute (HHMI), United States
  • 3University Medical Center Utrecht, Netherlands

All cells respond to extracellular signals by altering their intracellular biochemical state. In neurons, such signaling regulates many aspects of cell and synapse biology and induces changes that are thought to be important for nervous system development, its adaptation in the face of a changing environment, and ongoing homeostatic maintenance. Although great advances have been made in developing novel fluorescent reporters of intracellular signaling as well as in methods of fluorescence detection for use in freely moving animals, these approaches have generally not been combined. Thus, we know relatively little about how the intracellular biochemical state of neurons, and other cell classes, is dynamically regulated during animals’ behavior. Here we describe a single multi-mode fiber based fluorescent lifetime photometry system (FLiP) designed to monitor the state of fluorescence reporters of biochemical state in freely moving animals. We demonstrate the utility of FLiP by monitoring the lifetime of FLIM-AKAR, a genetically encoded fluorescent reporter of PKA phosphorylation, in populations of direct and indirect pathway striatal projection neurons in mice receiving food rewards. We find that the activity of PKA in each pathway is transiently regulated by reward acquisition, with PKA phosphorylation being enhanced and repressed in direct and indirect pathway neurons, respectively. This study demonstrates the power of FLiP to detect changes in biochemical state induced by naturalistic experiences in behaving animals.

Keywords: FLIM (fluorescence lifetime imaging microscopy), Fiber Photometry, PKA, Dopamine, Accumbens

Received: 06 Jun 2019; Accepted: 09 Jul 2019.

Edited by:

Keith Mathieson, University of Strathclyde, United Kingdom

Reviewed by:

Minmin Luo, Tsinghua University, China
Guohong Cui, National Institutes of Health (NIH), United States  

Copyright: © 2019 Sabatini, Lee, Chen and Lodder. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Dr. Bernardo Sabatini, Harvard Medical School, Boston, United States, bsabatini@hms.harvard.edu