Protocol to visualize trans-interaction of clustered protocadherin using cIPAD, a fluorescent indicator, in cultured human cells and mouse neurons

Abstract

Summary: cIPAD is a fluorescent indicator that allows the visualization of trans-interactions of clustered protocadherin (Pcdh), a cell adhesion molecule that mediates neuronal self-recognition. We describe steps for using HEK293T cells to visualize Pcdh trans-interactions across cells as a preliminary experiment before using dissociated mouse neurons. We then detail procedures for visualizing Pcdh trans-interactions between processes originating from the same neurons, which are considered as Pcdh-mediated neuronal self-recognition.For complete details on the use and execution of this protocol, please refer to Kanadome et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Biotechnology and bioengineering
• Cell Biology
• Neuroscience