Expanded huntingtin activates the c-Jun terminal kinase/c-Jun pathway prior to aggregate formation in striatal neurons in culture

Neuroscience. 2004;127(4):859-70. doi: 10.1016/j.neuroscience.2004.05.054.

Abstract

Huntington's disease (HD) is an autosomal neurodegenerative disorder, caused by expansion of a glutamine repeat in the Huntingtin protein. Pathogenesis in HD includes the cytoplasmic cleavage of Huntingtin and release of an amino-terminal fragment capable of nuclear localization, where expanded-Huntingtin (Exp-Htt) might lead to aberrant transcriptional regulation, neuronal dysfunction and degeneration. Recent evidence, from hippocampal cell lines, also implicates altered interaction of Exp-Htt with components of the c-Jun N-terminal kinase (JNK) cascade. However, there is yet no proven implication of the JNK/c-Jun module in degeneration of striatal neurons, the more vulnerable cell population, in HD. In the present study, we used primary striatal neurons in culture to analyze c-Jun activation by Exp-Htt. Green fluorescent protein (GFP)-tagged exon 1 of human Huntingtin either in its normal (25Q, normal-Htt) or expanded (103Q, Exp-Htt) version was transiently transfected in these cells. We first set out, in our conditions, the time course of striatal degeneration produced by Exp-Htt, and found it occurred rapidly. At 48 h post-transfection, 60% of striatal neurons expressing Exp-Htt had apoptotic characteristics including DNA fragmentation and neuritic retraction. Most of these neurons also showed nuclear aggregates of GFP-Exp Htt. Kinetics of c-Jun activation were tested in transfected cells using immunocytochemical detection of phospho-c-Jun. We found a significant activation and induction of c-Jun in Exp-Htt but not normal-Htt-transfected neurons. Of interest, these events occurred prior to nuclear translocation of Exp-Htt. Finally, overexpression of a dominant negative version of c-Jun, as well as pharmacological inhibition of JNK strongly protected against DNA fragmentation and neuritic retraction induced by Exp-Htt. Thus our data suggest that c-Jun activation and induction, is an early event in the pathogenesis of HD, occurring prior to formation of nuclear aggregates of Exp-Htt.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / physiology
  • Cells, Cultured
  • Cerebral Cortex / chemistry
  • Cerebral Cortex / enzymology
  • Cerebral Cortex / pathology
  • Corpus Striatum / chemistry
  • Corpus Striatum / cytology
  • Corpus Striatum / enzymology*
  • DNA Fragmentation / physiology
  • Embryo, Mammalian
  • Enzyme Activation / physiology
  • Female
  • Green Fluorescent Proteins
  • Huntingtin Protein
  • JNK Mitogen-Activated Protein Kinases
  • Luminescent Proteins / genetics
  • MAP Kinase Signaling System / physiology*
  • Mice
  • Mitogen-Activated Protein Kinases / analysis
  • Mitogen-Activated Protein Kinases / metabolism*
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism*
  • Nerve Tissue Proteins / physiology*
  • Neurons / chemistry
  • Neurons / enzymology*
  • Neurons / pathology
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • Nuclear Proteins / physiology*
  • Pregnancy

Substances

  • Htt protein, mouse
  • Huntingtin Protein
  • Luminescent Proteins
  • Nerve Tissue Proteins
  • Nuclear Proteins
  • Green Fluorescent Proteins
  • JNK Mitogen-Activated Protein Kinases
  • Mitogen-Activated Protein Kinases