Treatment of Rat Coronal Slices with a Neurotoxic Peptide

Take a rat coronal brain section and cut it along the midline to obtain hemislices, ensuring both originate from the same anatomical region for control and test experiments.

Transfer the hemislices to oxygenated aCSF for recovery.

Next, place them in vials with aCSF for the control and aCSF with a neurotoxic peptide for the test.

Seal the vials with oxygenating needles to supply oxygen to the tissue and incubate.

In the test vial, the neurotoxic peptide binds to its neuronal receptor, disrupting normal neuronal signaling.

This disrupted signaling leads to neurotoxic protein accumulation and alters microtubule-associated proteins, promoting neurodegeneration.

After incubation, transfer the hemislices to tubes containing chilled lysis buffer.

Homogenize the tissue to release cellular contents, while inhibitors in the buffer prevent protein degradation.

Centrifuge and collect the supernatant containing soluble proteins and store them for further analysis.

Carefully divide each section with small scissors along the midline to obtain two specular hemi-slices. Use each pair of hemi-slices for the control and treated condition. After being divided, let the hemi-sections rest in the vibrat