Transgenic silencing of neurons in the mammalian brain by expression of the allatostatin receptor (AlstR)

J Neurophysiol. 2009 Oct;102(4):2554-62. doi: 10.1152/jn.00480.2009. Epub 2009 Aug 19.

Abstract

The mammalian brain is an enormously complex set of circuits composed of interconnected neuronal cell types. The analysis of central neural circuits will be greatly served by the ability to turn off specific neuronal cell types while recording from others in intact brains. Because drug delivery cannot be restricted to specific cell types, this can only be achieved by putting "silencer" transgenes under the control of neuron-specific promoters. Towards this end we have created a line of transgenic mice putting the Drosophila allatostatin (AL) neuropeptide receptor (AlstR) under the control of the tetO element, thus enabling its inducible expression when crossed to tet-transactivator lines. Mammals have no endogenous AL or AlstR, but activation of exogenously expressed AlstR in mammalian neurons leads to membrane hyperpolarization via endogenous G-protein-coupled inward rectifier K(+) channels, making the neurons much less likely to fire action potentials. Here we show that this tetO/AlstR line is capable of broadly expressing AlstR mRNA in principal neurons throughout the forebrain when crossed to a commercially-available transactivator line. We electrophysiologically characterize this cross in hippocampal slices, demonstrating that bath application of AL leads to hyperpolarization of CA1 pyramidal neurons, making them refractory to the induction of action potentials by injected current. Finally, we demonstrate the ability of AL application to silence the sound-evoked spiking responses of auditory cortical neurons in intact brains of AlstR/tetO transgenic mice. When crossed to other transactivator lines expressing in defined neuronal cell types, this AlstR/tetO line should prove a very useful tool for the analysis of intact central neural circuits.

Publication types

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

MeSH terms

  • Acoustic Stimulation
  • Action Potentials / physiology
  • Animals
  • Auditory Perception / physiology
  • CA1 Region, Hippocampal / physiology
  • Drosophila
  • Drosophila Proteins / genetics*
  • Drosophila Proteins / metabolism*
  • Evoked Potentials, Auditory / physiology
  • Gene Silencing*
  • In Vitro Techniques
  • Membrane Potentials / physiology
  • Mice
  • Mice, Transgenic
  • Neurons / physiology*
  • Neuropeptides / metabolism
  • Potassium Channels, Inwardly Rectifying / metabolism
  • Prosencephalon / physiology*
  • Pyramidal Cells / physiology
  • RNA, Messenger / metabolism
  • Receptors, G-Protein-Coupled / genetics*
  • Receptors, G-Protein-Coupled / metabolism*
  • Receptors, Neuropeptide / genetics*
  • Receptors, Neuropeptide / metabolism*

Substances

  • AstA-R1 protein, Drosophila
  • Drosophila Proteins
  • Neuropeptides
  • Potassium Channels, Inwardly Rectifying
  • RNA, Messenger
  • Receptors, G-Protein-Coupled
  • Receptors, Neuropeptide
  • allatostatin