Mass spectrometric analysis of single identified neurons of an insect

Biochem Biophys Res Commun. 2005 Feb 18;327(3):640-5. doi: 10.1016/j.bbrc.2004.12.086.

Abstract

The combination of retrograde labelling with dextran-tetramethylrhodamine and MALDI-TOF mass spectrometry was used to analyse for the first time the peptidome of a series of morphologically identified single neurosecretory cells of an insect. Eight postero-lateral cells of the metathoracic ganglion of the American cockroach, Periplaneta americana, were used to demonstrate that: (1) the complete dissection procedure can be documented and (2) the mass spectrometric analysis of the dissected somata results in highly reproducible mass spectra. In total, 21 FMRFamide-related peptides were detected in each of the postero-lateral cells which release their neurosecretions via thoracic perisympathetic organs. Direct analysis of these neurohemal organs confirmed the co-storage of FMRFamide-related peptides. Two additional abundant peptides from thoracic perisympathetic organs which were not detectable in the postero-lateral cells were characterized using ESI-Q-TOF MS/MS. De novo sequencing yielded two related peptides (FERL/IEamides) without any similarity with known peptide families of insects.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cockroaches
  • FMRFamide / analysis
  • FMRFamide / chemistry
  • FMRFamide / metabolism
  • Ganglia, Invertebrate / cytology
  • Ganglia, Parasympathetic / chemistry
  • Ganglia, Parasympathetic / metabolism
  • Insecta
  • Neurons / chemistry*
  • Neurons / metabolism
  • Neuropeptides / analysis
  • Neuropeptides / chemistry
  • Neuropeptides / metabolism
  • Spectrometry, Mass, Electrospray Ionization / methods
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods

Substances

  • Neuropeptides
  • FMRFamide