Visualizing and Characterizing Semaphorin Endocytic Events Using Quantum Dot-Conjugated Proteins

Methods Mol Biol. 2017;1493:277-286. doi: 10.1007/978-1-4939-6448-2_20.

Abstract

Neurons can endocytose soluble semaphorins to either initiate or interrupt signaling at the cell membrane. Depending on the cell type and even on the specific subcellular domain, the endocytic process will differ in intensity, speed, and modality, and will subsequently facilitate diverse actions of semaphorin molecules. Therefore, in order to understand the physiology of guidance cues like semaphorins it is important to visualize endocytic events with good spatial and temporal resolution. Here, we describe methods to visualize endocytosed Semaphorin3A (Sema3A) molecules and to characterize the rate and pathway of internalization in primary rat neuronal cultures using semiconductor quantum dot nanoparticles (Q-dots).

Keywords: Acid strip; Endocytosis; Microscopy; Q-dots; Semaphorin3A.

MeSH terms

  • Animals
  • Cells, Cultured
  • Endocytosis / physiology*
  • Humans
  • Microscopy, Confocal
  • Nerve Tissue Proteins / metabolism*
  • Quantum Dots*
  • Rats
  • Semaphorin-3A / physiology*

Substances

  • Nerve Tissue Proteins
  • Semaphorin-3A