Improved dynamic monitoring of transcriptional activity during longitudinal analysis in the mouse brain

Hwon Heo, Juyeong Jo, Jae In Jung, Young Min Han, Seongsoo Lee, Song Rae Kim, Seung Hae Kwon, Kil Nam Kim, Byung Joon Hwang, Yun Kee, Byoung Dae Lee, Dongmin Kang, Song Her

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Bioluminescence imaging has proven to be a highly sensitive technique for assessing in vitro transcriptional activity toward understanding gene regulation patterns; however, application of this technique is limited for brain research. In particular, the poor spatiotemporal resolution is a major hurdle for monitoring the dynamic changes of transcriptional activity in specific regions of the brain during longitudinal analysis of living animals. To overcome this limitation, in this study, we modified a lentivirus-based luciferase glucocorticoid receptor (GR) reporter by inserting destabilizing sequence genes, and then the reporter was stereotaxically injected in the mouse infralimbic prefrontal cortex (IL-PFC). Using this strategy, we could successfully pin-point and monitor the dynamic changes in GR activity in IL-PFC during normal stress adaptation. The modified reporter showed a 1.5-fold increase in temporal resolution for monitoring GR activity compared to the control, with respect to the intra-individual coefficients of variation. This novel in vivo method has broad applications, as it is readily adaptable to different types of transcription factor arrays as well spanning wide target regions of the brain to other organs and tissues.

Original languageEnglish
Article numberbio037168
JournalBiology Open
Volume8
Issue number1
DOIs
StatePublished - 2019

Bibliographical note

Funding Information:
This research was supported by the Brain Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning [NRF-2016M3C7A1905383].

Publisher Copyright:
© 2019. Published by The Company of Biologists Ltd.

Keywords

  • Brain promoter activity
  • Glucocorticoid receptor signaling
  • Longitudinal monitoring
  • Luciferase
  • Pin-point monitoring transcription activity
  • Stress

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