Mass Spectrometry Analysis of Wild-Type and Knock-in Q140/Q140 Huntington's Disease Mouse Brains Reveals Changes in Glycerophospholipids Including Alterations in Phosphatidic Acid and Lyso-Phosphatidic Acid

J Huntingtons Dis. 2015;4(2):187-201. doi: 10.3233/JHD-150149.

Abstract

Background: Huntington's disease (HD) is a neurodegenerative disease caused by a CAG expansion in the HD gene, which encodes the protein Huntingtin. Huntingtin associates with membranes and can interact directly with glycerophospholipids in membranes.

Objective: We analyzed glycerophospholipid profiles from brains of 11 month old wild-type (WT) and Q140/Q140 HD knock-in mice to assess potential changes in glycerophospholipid metabolism.

Methods: Polar lipids from cerebellum, cortex, and striatum were extracted and analyzed by liquid chromatography and negative ion electrospray tandem mass spectrometry analysis (LC-MS/MS). Gene products involved in polar lipid metabolism were studied using western blotting, immuno-electron microscopy and qPCR.

Results: Significant changes in numerous species of glycerophosphate (phosphatidic acid, PA) were found in striatum, cerebellum and cortex from Q140/Q140 HD mice compared to WT mice at 11 months. Changes in specific species could also be detected for other glycerophospholipids. Increases in species of lyso-PA (LPA) were measured in striatum of Q140/Q140 HD mice compared to WT. Protein levels for c-terminal binding protein 1 (CtBP1), a regulator of PA biosynthesis, were reduced in striatal synaptosomes from HD mice compared to wild-type at 6 and 12 months. Immunoreactivity for CtBP1 was detected on membranes of synaptic vesicles in striatal axon terminals in the globus pallidus.

Conclusions: These novel results identify a potential site of molecular pathology caused by mutant Huntingtin that may impart early changes in HD.

Keywords: CtBP; Huntingtin; Huntington’s disease; LPA; LPAAT; acyl transferase; glycerophospholipid; lipidomics; mass spectrometry; phosphatidic acid; phospholipase.

Publication types

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

MeSH terms

  • Alcohol Oxidoreductases / metabolism
  • Animals
  • Brain / metabolism*
  • Brain / ultrastructure
  • Cerebellum / metabolism
  • Cerebral Cortex / metabolism
  • Corpus Striatum / metabolism
  • Corpus Striatum / ultrastructure
  • DNA-Binding Proteins / metabolism
  • Disease Models, Animal
  • Gene Knock-In Techniques
  • Glycerophospholipids / metabolism*
  • Huntingtin Protein
  • Huntington Disease / genetics
  • Huntington Disease / metabolism*
  • Lysophospholipids / metabolism*
  • Male
  • Mass Spectrometry
  • Mice
  • Mice, Inbred C57BL
  • Nerve Tissue Proteins / genetics
  • Neurons / metabolism
  • Neurons / ultrastructure
  • Phosphatidic Acids / metabolism*
  • Synaptic Vesicles / metabolism
  • Synaptic Vesicles / ultrastructure
  • Synaptosomes / metabolism

Substances

  • DNA-Binding Proteins
  • Glycerophospholipids
  • HTT protein, human
  • Huntingtin Protein
  • Lysophospholipids
  • Nerve Tissue Proteins
  • Phosphatidic Acids
  • Alcohol Oxidoreductases
  • C-terminal binding protein
  • lysophosphatidic acid