hAChE-IN-5

This protocol assesses in vitro neurotoxicity by combining neuronal viability, mitochondrial/metabolic activity, neurite outgrowth, and optional neuronal network function readouts. Calcein-AM or resazurin/PrestoBlue readouts estimate viable or metabolically active cells; βIII-tubulin immunofluorescence detects neuronal morphology and neurite networks; TMRE detects mitochondrial membrane potential; and MEA recordings detect functional changes in neuronal network activity.

Cytotoxicity assays are usually based on the assessment of cell membrane damage, which can also be indirectly detected by measuring cell viability. Detection methods include MTT assay, CKK-8 assay, LDH assay and ATP assay, etc.

Periodic acid-Schiff staining detects tissue carbohydrates and mucosubstances by oxidizing carbohydrate glycol groups with periodic acid to generate aldehydes, which then react with Schiff reagent to produce a magenta reaction product; classic reports established the method for mucin and polysaccharide-containing structures in fixed tissue sections. PAS staining can demonstrate neutral mucins and goblet-cell mucin, but it is not specific for mucin because glycogen and other PAS-positive tissue components can also stain; diastase/PAS-D is used when glycogen removal is needed to distinguish glycogen-dependent PAS signal from non-glycogen PAS-positive mucosubstances.

Alzheimer’s Disease (AD) is a neurodegenerative disorder characterized by a progressive decline in cognitive functions and loss of specific types of neurons and synapses. Alzheimer's symptoms can be simulated in mice by injecting drugs (such as Aβ) or genetically modified.