Visualizing the Synaptogenesis of Cerebellar Granule Neurons at Different Developmental Stages

Begin with pre-fixed brain tissue slices collected from early and late postnatal stages of mice.

The slice contains cerebellar granule neurons, or CGNs, expressing a fluorescent protein.

Using a confocal microscope, focus on a single fluorescent CGN.

Capture z-stack images at varying depths to create a three-dimensional view of the neuron.

Using imaging software, trace the neurons' neurites or long projections, including dendrites and axons.

Measure dendrite length from the cell body to its tip.

Analyze dendritic claw formation, specialized structures at dendrite tips where signals from other neurons are received.

Also, assess the neuron's surface area and volume.

In the early postnatal stage, neurons show an increased number of dendrites, with no claw formation.

Meanwhile, in the late postnatal stage, there are fewer dendrites, but their lengths and the number of claws increase.

Thus, with age, CGNs undergo synaptogenesis and form connections with other cells for signal transmission.

To study the morphology of single electroporated CGNs from sagittal brain sections of the experimental pup, take z stack images at 0.5 micrometers per stack on a confocal microscope. Image one cell per image