Fluorescent peptide-PNA chimeras for imaging monoamine oxidase A mRNA in neuronal cells

Bioconjug Chem. 2012 Feb 15;23(2):158-63. doi: 10.1021/bc2004507. Epub 2012 Jan 25.

Abstract

Monoamine oxidases (MAO) catalyze the oxidative deamination of many biogenic amines and are integral proteins found in the mitochondrial outer membrane. Changes in MAO-A levels are associated with depression, trait aggression, and addiction. Here we report the synthesis, characterization, and in vitro evaluation of novel fluorescent peptide-peptide nucleic acid (PNA) chimeras for MAOA mRNA imaging in live neuronal cells. The probes were designed to include MAOA-specific PNA dodecamers, separated by an N-terminal spacer to a μ-opioid receptor targeting peptide (DAMGO), with a spacer and a fluorophore on the C-terminus. The probe was successfully delivered into human SH-SY5Y neuroblastoma cells through μ-opioid receptor-mediated endocytosis. The K(d) by flow cytometry was 11.6 ± 0.8 nM. Uptake studies by fluorescence microscopy showed ∼5-fold higher signal in human SH-SY5Y neuroblastoma cells than in negative control CHO-K1 cells that lack μ-opioid receptors. Moreover, a peptide-mismatch control sequence showed no significant uptake in SH-SY5Y cells. Such mRNA imaging agents with near-infrared fluorophores might enable real time imaging and quantitation of neuronal mRNAs in live animal models.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • CHO Cells
  • Cricetinae
  • Flow Cytometry
  • Fluorescent Dyes / analysis*
  • Fluorescent Dyes / chemistry
  • Humans
  • Microscopy, Fluorescence
  • Molecular Imaging*
  • Molecular Structure
  • Monoamine Oxidase / analysis
  • Monoamine Oxidase / genetics*
  • Neuroblastoma / enzymology
  • Neuroblastoma / genetics
  • Neurons / metabolism*
  • Peptide Nucleic Acids / analysis*
  • Peptide Nucleic Acids / chemistry
  • Peptides / analysis*
  • Peptides / chemistry
  • RNA, Messenger / analysis*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism

Substances

  • Fluorescent Dyes
  • Peptide Nucleic Acids
  • Peptides
  • RNA, Messenger
  • Monoamine Oxidase