PACα--an optogenetic tool for in vivo manipulation of cellular cAMP levels, neurotransmitter release, and behavior in Caenorhabditis elegans

J Neurochem. 2011 Feb;116(4):616-25. doi: 10.1111/j.1471-4159.2010.07148.x. Epub 2011 Jan 20.

Abstract

Photoactivated adenylyl cyclase α (PACα) was originally isolated from the flagellate Euglena gracilis. Following stimulation by blue light it causes a rapid increase in cAMP levels. In the present study, we expressed PACα in cholinergic neurons of Caenorhabditis elegans. Photoactivation led to a rise in swimming frequency, speed of locomotion, and a decrease in the number of backward locomotion episodes. The extent of the light-induced behavioral effects was dependent on the amount of PACα that was expressed. Furthermore, electrophysiological recordings from body wall muscle cells revealed an increase in miniature post-synaptic currents during light stimulation. We conclude that the observed effects were caused by cAMP synthesis because of photoactivation of pre-synaptic PACα which subsequently triggered acetylcholine release at the neuromuscular junction. Our results demonstrate that PACα can be used as an optogenetic tool in C. elegans for straightforward in vivo manipulation of intracellular cAMP levels by light, with good temporal control and high cell specificity. Thus, using PACα allows manipulation of neurotransmitter release and behavior by directly affecting intracellular signaling.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcholine / physiology
  • Adenylyl Cyclases / biosynthesis
  • Adenylyl Cyclases / metabolism
  • Adenylyl Cyclases / physiology*
  • Animals
  • Animals, Genetically Modified
  • Caenorhabditis elegans / enzymology*
  • Caenorhabditis elegans Proteins / biosynthesis
  • Caenorhabditis elegans Proteins / metabolism
  • Caenorhabditis elegans Proteins / physiology*
  • Cyclic AMP / biosynthesis
  • Cyclic AMP / metabolism*
  • Euglena gracilis / enzymology
  • Motor Activity / physiology*
  • Neurons / enzymology
  • Neurons / metabolism*
  • Neurotransmitter Agents / metabolism*
  • Photic Stimulation / methods

Substances

  • Caenorhabditis elegans Proteins
  • Neurotransmitter Agents
  • Cyclic AMP
  • Adenylyl Cyclases
  • Acetylcholine