Calcium Imaging of Enteric Neuron and Glia Activities in a Mouse Colonic Myenteric Plexus

Take an imaging chamber containing immobilized mouse colonic myenteric plexus tissue, which houses ganglionated plexuses composed of networks of enteric neurons and glial cells.

These cells are labeled with a calcium indicator that fluoresces upon calcium binding.

Next, place the chamber on a fluorescence microscope stage. Perfuse the chamber with a pre-warmed buffer containing inhibitors to suppress muscle contractions during imaging.

Using a microscope,  identify healthy ganglionic regions that exhibit uniform fluorescence.

Acquire fluorescent images to measure baseline activity, providing a reference for intracellular calcium levels.

Perfuse the tissue with a receptor agonist to activate neuronal receptors, which triggers intracellular calcium influx and an increase in fluorescence intensity.

Neuronal activation indirectly stimulates surrounding glial cells, leading to calcium influx and a subsequent increase in fluorescence.

Capture time-lapse fluorescent images to measure changes in fluorescence intensity, which indicate calcium dynamics and signaling activity in both neurons and glial cells relative to baseline levels.