Standardized protocol for plasticity assessment in the aging mouse neocortex using choline-chloride perfusion.

Area of Science:

• Neuroscience
• Aging Research
• Cellular Biology

Background:

• Age-related synaptic dysfunction contributes to neurodegeneration.
• Preparing high-quality brain tissue from aged animals for analysis is difficult.
• Existing methods struggle to preserve neuronal structure and function in aging brains.

Purpose of the Study:

• To develop a standardized protocol for preparing intact acute brain slices from mice of different ages.
• To enable combined structural and functional analysis of individual neurons across the lifespan.
• To investigate age-related changes in synaptic transmission and plasticity.

Main Methods:

• Standardized acute brain slice preparation using transcardial choline-chloride perfusion.
• Analysis of cortical lamination and subcellular synaptic structure in medial prefrontal cortex (mPFC) pyramidal neurons.
• Whole-cell patch-clamp recordings to examine spontaneous excitatory synaptic transmission and forskolin-induced chemical long-term potentiation (cLTP).

Main Results:

• The protocol reliably yields intact cortical slices from young and aged mice.
• Cortical lamination and synaptic structure are preserved in layer 2/3 pyramidal neurons.
• Synaptic plasticity, measured by cLTP, shows age-related differences in expression.

Conclusions:

• The developed protocol is a reproducible framework for studying synaptic transmission and plasticity in the aging cortex.
• This method is broadly applicable to research on age-related brain disorders and neurodegeneration.
• The findings highlight age-dependent alterations in synaptic plasticity mechanisms.