Effective expression of the green fluorescent fusion proteins in cultured Aplysia neurons

I. Sahly, H. Erez, A. Khoutorsky, E. Shapira, Micha E. Spira*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

25 Scopus citations

Abstract

The green fluorescent fusion protein and its isoforms are extensively used to monitor gene expression, protein localisation and their dynamics in relations to fundamental cellular processes. However, it has not yet been effectively applied to Aplysia neurons that serve as a powerful model to study the mechanisms underlying neuroplasticity. We report here the development of a procedure combining in vitro transcription of mRNA encoding fluorescent-tagged proteins and its subsequent injection into the cytoplasm to image, in real-time, protein dynamics in cultured Aplysia neurones. To illustrate the efficiency of the procedure we report here the visualisation of actin, microtubules and vesicle trafficking. The results presented here introduce a reliable and effective method to express green fluorescent protein (GFP) fusion proteins in cultured Aplysia neurons.

Original languageAmerican English
Pages (from-to)111-117
Number of pages7
JournalJournal of Neuroscience Methods
Volume126
Issue number2
DOIs
StatePublished - 30 Jun 2003

Bibliographical note

Funding Information:
This study was supported by grants from the Israel Science Foundation (No. 620/98), the US-IsraelBi-National Research Foundation (No. 97-00297-1) and the Charles E. Smith Family Laboratory for Collaborative Research in Psychobiology, the Hebrew University of Jerusalem. M.E. Spira is the Levi DeViali Professor in Neurobiology.

Keywords

  • Actin
  • Aplysia neurons
  • GFP
  • Green fluorescent protein
  • Tubulin
  • mRNA

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