Proteomics analysis of perilymph and cerebrospinal fluid in mouse

Laryngoscope. 2009 May;119(5):953-8. doi: 10.1002/lary.20209.

Abstract

Objectives: Proteins in perilymph may alter the delivery profile of implantable intracochlear drug delivery systems through biofouling. Knowledge of protein composition will help anticipate interactions with delivered agents.

Study design: Analysis of mouse perilymph.

Methods: Protein composition of perilymph and cerebrospinal fluid (CSF) was analyzed using a capillary liquid chromatography-mass spectrometry-based iTRAQ quantitative proteomics approach. We searched against a mouse subset of the Uniprot FASTA protein database. We sampled perilymph from the apex of the mouse cochlea to minimize CSF contamination.

Results: More than 50 explicit protein isoforms were identified with very high confidence. iTRAQ reporter ions allowed determination of relative molar amounts of proteins between perilymph and CSF. Protein in perilymph was almost three times more concentrated than in CSF. More than one-third of the proteins in perilymph comprised protease inhibitors, with serpins being the predominant group. Apolipoproteins constituted 16%. Fifteen percent of the proteins were enzymes. Albumin was the most abundant single protein (14%). Proteins with relatively high perilymph/CSF ratios included broad-spectrum protease inhibitors and apolipoproteins.

Discussion: Some proteins found in perilymph, such as albumin and HMW kininogen, have been implicated in biofouling through adsorption to device materials. The relatively large quantities of apolipoprotein and albumin may serve as a reservoir for acidic and lipophilic drugs. Alpha-2-glycoprotein can bind basic drugs.

Conclusions: Perilymph is similar in protein composition to CSF, though amounts are 2.8 times higher. Protease inhibitors comprise the largest category of proteins.

Publication types

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

MeSH terms

  • Animals
  • Cerebrospinal Fluid / chemistry*
  • Chromatography, Liquid
  • Male
  • Mass Spectrometry
  • Mice
  • Perilymph / chemistry*
  • Proteomics*