Computational protocol: Inhibition of the Polyamine System Counteracts β-Amyloid Peptide-Induced Memory Impairment in Mice: Involvement of Extrasynaptic NMDA Receptors

Similar protocols

Protocol publication

[…] Images were acquired using a confocal microscope (Leica TCS-SP5-II. Leica-Microsystems, Mannheim, Germany) equipped with Plan Apo 63× oil NA 1.4 objective and a Diode (405 nm), Argon (458, 476, 488, 496, 514 nm laser lines), Diode Pumped Solid State (DPSS, 561 nm) and HeNe (633 nm) lasers. Laser intensity and signal detection settings were held constant to allow quantitative comparison between experimental groups. Cells were scanned with optical serial sections of 0.29 µm intervals and, depending on the experiment, maximum intensity or mean average projection of z-stack images from individual cell nuclei and dendritic branches were generated (ImageJ software, NIH, Bethesda, USA).For dendritic spines density and morphology experiments, one to three dendritic segments (20 µm), distal and proximal from soma of each neuron were used, and analysis was performed using NeuronStudio software 0.9.92 . Spines were defined as protrusions that could be differentiated from the dendritic shaft and restricted to those that were visible in the x- and y-axes.Quantification of nuclear accumulation of Jacob was performed as previously described . Briefly, images were open on Image J software and the nuclear region of interest (ROI) was defined using the threshold from the DAPI staining. Nuclear Jacob immunoreactivity was measured as mean grey values (arbitrary units in pixel intensity). Data plotted in the graphs were normalized relative to control group. […]

Pipeline specifications

Software tools ImageJ, NeuronStudio
Applications Laser scanning microscopy, Microscopic phenotype analysis
Organisms Mus musculus
Diseases Alzheimer Disease, Liver Diseases
Chemicals Polyamines, Spermidine, Spermine