References of "Journal of Neuroscience Methods"
     in
Bookmark and Share    
Peer Reviewed
See detailQuantitative quadruple-label immunofluorescence of mitochondrial and cytoplasmic proteins in single neurons from human midbrain tissue.
Grünewald, Anne UL; Lax, Nichola Z.; Rocha, Mariana C. et al

in Journal of Neuroscience Methods (2014), 232

BACKGROUND: Respiratory chain (RC) deficiencies are found in primary mtDNA diseases. Focal RC defects are also associated with ageing and neurodegenerative disorders, e.g. in substantia nigra (SN) neurons ... [more ▼]

BACKGROUND: Respiratory chain (RC) deficiencies are found in primary mtDNA diseases. Focal RC defects are also associated with ageing and neurodegenerative disorders, e.g. in substantia nigra (SN) neurons from Parkinson's disease patients. In mitochondrial disease and ageing, mtDNA mutational loads vary considerably between neurons necessitating single cell-based assessment of RC deficiencies. Evaluating the full extent of RC deficiency within SN neurons is challenging because their size precludes investigations in serial sections. We developed an assay to measure RC abnormalities in individual SN neurons using quadruple immunofluorescence. NEW METHOD: Using antibodies against subunits of complex I (CI) and IV, porin and tyrosine hydroxylase together with IgG subtype-specific fluorescent labelled secondary antibodies, we quantified the expression of CI and CIV compared to mitochondrial mass in dopaminergic neurons. CI:porin and CIV:porin ratios were determined relative to a standard control. RESULTS: Quantification of expression of complex subunits in midbrain sections from patients with mtDNA disease and known RC deficiencies consistently showed reduced CI:porin and/or CIV:porin ratios. COMPARISON WITH EXISTING METHOD(S): The standard histochemical method to investigate mitochondrial dysfunction, the cytochrome c oxidase/succinate dehydrogenase assay, measures CIV and CII activities. To also study CI in a patient, immunohistology in additional sections, i.e. in different neurons, is required. Our method allows correlation of the expression of CI, CIV and mitochondrial mass at a single cell level. CONCLUSION: Quantitative quadruple-label immunofluorescence is a reliable tool to measure RC deficiencies in individual neurons that will enable new insights in the molecular mechanisms underlying inherited and acquired mitochondrial dysfunction. [less ▲]

Detailed reference viewed: 124 (6 UL)
Full Text
Peer Reviewed
See detailAn efficient method to limit microglia-dependent effects in astroglial cultures.
Losciuto, Sophie; Dorban, Gauthier; Gabel, Sébastien et al

in Journal of Neuroscience Methods (2012), 207(1), 59-71

Detailed reference viewed: 113 (6 UL)
Full Text
Peer Reviewed
See detailAutomated analysis of neuronal morphology, synapse number and synaptic recruitment.
Schmitz, Sabine UL; Hjorth, J. J. Johannes; Joemai, Raoul M. S. et al

in Journal of Neuroscience Methods (2011), 195(2), 185-93

The shape, structure and connectivity of nerve cells are important aspects of neuronal function. Genetic and epigenetic factors that alter neuronal morphology or synaptic localization of pre- and post ... [more ▼]

The shape, structure and connectivity of nerve cells are important aspects of neuronal function. Genetic and epigenetic factors that alter neuronal morphology or synaptic localization of pre- and post-synaptic proteins contribute significantly to neuronal output and may underlie clinical states. To assess the impact of individual genes and disease-causing mutations on neuronal morphology, reliable methods are needed. Unfortunately, manual analysis of immuno-fluorescence images of neurons to quantify neuronal shape and synapse number, size and distribution is labor-intensive, time-consuming and subject to human bias and error. We have developed an automated image analysis routine using steerable filters and deconvolutions to automatically analyze dendrite and synapse characteristics in immuno-fluorescence images. Our approach reports dendrite morphology, synapse size and number but also synaptic vesicle density and synaptic accumulation of proteins as a function of distance from the soma as consistent as expert observers while reducing analysis time considerably. In addition, the routine can be used to detect and quantify a wide range of neuronal organelles and is capable of batch analysis of a large number of images enabling high-throughput analysis. [less ▲]

Detailed reference viewed: 98 (3 UL)
Peer Reviewed
See detailA simple method for a specific retrograde labelling of dorsal root and sympathetic ganglion cells innervating the knee joint of the cat.
Hanesch, Ulrike UL; Heppelmann, Bernd

in Journal of Neuroscience Methods (1995), 63

Detailed reference viewed: 65 (0 UL)