A brain-sparing diphtheria toxin for chemical genetic ablation of peripheral cell lineages

Nat Commun. 2017 Apr 3;8:14967. doi: 10.1038/ncomms14967.

Abstract

Conditional expression of diphtheria toxin receptor (DTR) is widely used for tissue-specific ablation of cells. However, diphtheria toxin (DT) crosses the blood-brain barrier, which limits its utility for ablating peripheral cells using Cre drivers that are also expressed in the central nervous system (CNS). Here we report the development of a brain-sparing DT, termed BRAINSPAReDT, for tissue-specific genetic ablation of cells outside the CNS. We prevent blood-brain barrier passage of DT through PEGylation, which polarizes the molecule and increases its size. We validate BRAINSPAReDT with regional genetic sympathectomy: BRAINSPAReDT ablates peripheral but not central catecholaminergic neurons, thus avoiding the Parkinson-like phenotype associated with full dopaminergic depletion. Regional sympathectomy compromises adipose tissue thermogenesis, and renders mice susceptible to obesity. We provide a proof of principle that BRAINSPAReDT can be used for Cre/DTR tissue-specific ablation outside the brain using CNS drivers, while consolidating the link between adiposity and the sympathetic nervous system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / cytology*
  • Cell Lineage*
  • Diphtheria Toxin / chemistry
  • Diphtheria Toxin / metabolism*
  • Disease Susceptibility
  • Energy Metabolism
  • Feeding Behavior
  • Female
  • Gene Deletion*
  • Gene Expression Regulation
  • Glucose Intolerance / complications
  • Glucose Intolerance / pathology
  • HeLa Cells
  • Humans
  • Hydrodynamics
  • Male
  • Mice
  • Movement
  • Neurons / metabolism
  • Norepinephrine / metabolism
  • Obesity / complications
  • Obesity / pathology
  • Parkinson Disease / pathology
  • Parkinson Disease / physiopathology
  • Polyethylene Glycols / chemistry
  • Sympathectomy
  • Thermogenesis / genetics

Substances

  • Diphtheria Toxin
  • Polyethylene Glycols
  • Norepinephrine