Morphometric Analysis in Neurodegenerative Disorders

Abstract

The study of dendritic length and spine density has become a standard in the analysis of neuronal abnormalities since a considerable number of neurological diseases have their foundation in alterations in these structures. One of the best ways to study possible alterations in neuronal morphometry is the use of Golgi impregnation. Introduced more than a century ago, it is still the standard and state-of-the-art technique for visualization of neuronal architecture. We successfully applied the Golgi method to mouse, rat, monkey and human brain tissues for studying both the normal and abnormal morphology of neurons. We were able to discover subtle morphological alterations in neuronal dendrites and dendritic spines in different brain areas. Although Golgi preparations can be examined by electronic microscopy, we used light microscopy and Neurolucida reconstruction to quantitatively explore the relationship between total dendritic length and spine density in different types of neurons. This review summarizes the methodology used to quantify neuronal abnormalities and discusses the utility of these techniques in different models of neurodegeneration.

Dietary Intake of Sulforaphane-Rich Broccoli Sprout Extracts during Juvenile and Adolescence Can Prevent Phencyclidine-Induced Cognitive Deficits at Adulthood

“…Briefly, images of dendrites within medial prefrontal cortex (mPFC), hippocampal CA1, CA3, and dentate gyrus (DG), nucleus accumbens (NAc)-core, NAc-shell, striatum and ventral tegmental area (VTA) were captured using a 100× objective with a Keyence BZ-9000 GenerationⅡmicroscope (Osaka, Japan). Spine density in these regions was counted as previously reported [ 29 – 31 ]. For spine density measurements, all clearly evaluable areas containing 50–100 μm of secondary dendrites from each imaged neuron were used.…”

Antidepressant Effects of TrkB Ligands on Depression-Like Behavior and Dendritic Changes in Mice After Inflammation

“…Briefly, images of dendrites within CA1, CA3, and DG of the hippocampus, PFC and NAc were captured using a 100× objective with a Keyence BZ-9000 GenerationⅡmicroscope (Osaka, Japan). Spines were counted along CA1, CA3, DG, PFC and NAc dendrites starting from their point of origin from the primary dendrite, as previously reported ( Milatovic et al, 2010 ). For spine density measurements, all clearly evaluable areas containing 50-100 µm of secondary dendrites from each imaged neuron were used.…”

Effects of prenatal binge‐like ethanol exposure and maternal stress on postnatal morphological development of hippocampal neurons in rats

“…The rapid Golgi staining was performed by using the FD Rapid GolgiStain™ kit. After impregnating and fixing the tissues for approximately 2 weeks, the brain specimens were cut into 100 μm sections (Lawrence et al, 2012;Milatovic et al, 2010;Min et al, 2016) on Leica cryostat at −22°C. Following drying, the sections were stained with appropriate solutions of the kit and counterstained with cresyl violet.…”