All-optical presynaptic plasticity induction by photoactivated adenylyl cyclase targeted to axon terminals

Cell Rep Methods. 2024 Apr 22;4(4):100740. doi: 10.1016/j.crmeth.2024.100740. Epub 2024 Mar 22.

Abstract

Intracellular signaling plays essential roles in various cell types. In the central nervous system, signaling cascades are strictly regulated in a spatiotemporally specific manner to govern brain function; for example, presynaptic cyclic adenosine monophosphate (cAMP) can enhance the probability of neurotransmitter release. In the last decade, channelrhodopsin-2 has been engineered for subcellular targeting using localization tags, but optogenetic tools for intracellular signaling are not well developed. Therefore, we engineered a selective presynaptic fusion tag for photoactivated adenylyl cyclase (bPAC-Syn1a) and found its high localization at presynaptic terminals. Furthermore, an all-optical electrophysiological method revealed rapid and robust short-term potentiation by bPAC-Syn1a at brain stem-amygdala synapses in acute brain slices. Additionally, bPAC-Syn1a modulated mouse immobility behavior. These results indicate that bPAC-Syn1a can manipulate presynaptic cAMP signaling in vitro and in vivo. The all-optical manipulation technique developed in this study can help further elucidate the dynamic regulation of various cellular functions.

Keywords: CP: neuroscience; all-optical manipulation; intracellular signaling; mouse; optogenetics; presynaptic targeting.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenylyl Cyclases* / genetics
  • Adenylyl Cyclases* / metabolism
  • Animals
  • Cyclic AMP* / metabolism
  • HEK293 Cells
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Neuronal Plasticity* / physiology
  • Optogenetics / methods
  • Presynaptic Terminals* / metabolism
  • Rats

Substances

  • Adenylyl Cyclases
  • Cyclic AMP