A comparative study of Lrrk2 function in primary neuronal cultures

Justus C. Dächsel, Bahareh Behrouz, Mei Yue, Joel E. Beevers, Heather L Melrose, Matthew J. Farrer

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

Objective: To assess the contribution of wild-type, mutant and loss of leucine-rich repeat kinase-2 (LRRK2; Lrrk2) on dendritic neuronal arborization. Background: LRRK2 mutations are recognized as the major genetic determinant of susceptibility to Parkinson's disease for which a cellular assay of Lrrk2 mutant function would facilitate the development of targeted molecular therapeutics. Methods: Dendritic neuronal arborization (neurite length, branching and the number of processes per cell) was quantified in primary hippocampal and midbrain cultures derived from five lines of recombinant LRRK2 mice, including human BAC wild-type and mutant overexpressors (Y1699C and G2019S), murine knock-out and G2019S knock-in animals. Results: Neuronal arborization in cultures from BAC Lrrk2 wild-type animals is comparable to non-transgenic littermate controls, despite high levels of human transgene expression. In contrast, primary neurons from both BAC mutant overexpressors presented with significantly reduced neuritic outgrowth and branching, although the total number of processes per cell remained comparable. The mutant-specific toxic gain-of-function observed in cultures from BAC mutant mice may be partially rescued by staurosporine treatment, a non-specific kinase inhibitor. In contrast, neuronal arborization is far more extensive in neuronal cultures derived from murine knock-out mice that lack endogenous Lrrk2 expression. In Lrrk2 G2019S knock-in mice, arguably the most physiologically relevant system, neuritic arborization is not impaired. Conclusions: Impairment of neuritic arborization is an exaggerated, albeit mutant specific, consequence of Lrrk2 over-expression in primary cultures. The phenotype and assay described provides a means to develop therapeutic agents that modulate the toxic gain-of-function conferred by mutant Lrrk2.

Original languageEnglish (US)
Pages (from-to)650-655
Number of pages6
JournalParkinsonism and Related Disorders
Volume16
Issue number10
DOIs
StatePublished - Dec 2010

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Neuronal Plasticity
Poisons
Phosphotransferases
Wild Animals
Staurosporine
Neurites
Genetic Predisposition to Disease
Mesencephalon
Transgenes
Knockout Mice
Leucine
Parkinson Disease
Phenotype
Neurons
Mutation
Therapeutics

Keywords

  • Drug screening
  • LRRK2
  • Neuritic outgrowth
  • Parkinson's disease
  • PD

ASJC Scopus subject areas

  • Geriatrics and Gerontology
  • Clinical Neurology
  • Neurology

Cite this

Dächsel, J. C., Behrouz, B., Yue, M., Beevers, J. E., Melrose, H. L., & Farrer, M. J. (2010). A comparative study of Lrrk2 function in primary neuronal cultures. Parkinsonism and Related Disorders, 16(10), 650-655. https://doi.org/10.1016/j.parkreldis.2010.08.018

A comparative study of Lrrk2 function in primary neuronal cultures. / Dächsel, Justus C.; Behrouz, Bahareh; Yue, Mei; Beevers, Joel E.; Melrose, Heather L; Farrer, Matthew J.

In: Parkinsonism and Related Disorders, Vol. 16, No. 10, 12.2010, p. 650-655.

Research output: Contribution to journalArticle

Dächsel, JC, Behrouz, B, Yue, M, Beevers, JE, Melrose, HL & Farrer, MJ 2010, 'A comparative study of Lrrk2 function in primary neuronal cultures', Parkinsonism and Related Disorders, vol. 16, no. 10, pp. 650-655. https://doi.org/10.1016/j.parkreldis.2010.08.018
Dächsel, Justus C. ; Behrouz, Bahareh ; Yue, Mei ; Beevers, Joel E. ; Melrose, Heather L ; Farrer, Matthew J. / A comparative study of Lrrk2 function in primary neuronal cultures. In: Parkinsonism and Related Disorders. 2010 ; Vol. 16, No. 10. pp. 650-655.
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