Imaging calcium dynamics in the nervous system by means of ballistic delivery of indicators

Petronella Kettunen, Jay Demas, Christian Lohmann, Narayanan Kasthuri, Yandao Gong, Rachel O.L. Wong, Wen Biao Gan

Research output: Contribution to journalArticlepeer-review

49 Scopus citations

Abstract

The use of fluorescence-based calcium indicators has, over the years, unraveled important calcium-dependent mechanisms underlying neuronal function and development. However, difficulties associated with the loading of calcium indicators have limited their widespread use, particularly for the study of neuronal processing in the adult nervous system. Here, we show that in the central and peripheral nervous systems, populations of neurons and their processes, including dendritic spines and filopodia, can be labeled rapidly and efficiently by delivering calcium indicator-coated particles using a 'gene gun'. Importantly, neuronal labeling occurred both in vitro and in vivo, and across a wide range of ages and preparations. The labeled cells demonstrate spontaneous and evoked calcium transients, indicating that particle-mediated delivery is not deleterious to neuronal function. Furthermore, unlike loading with patch pipettes, cytoplasmic content is preserved following ballistic loading. This enables the study of calcium-dependent second messenger pathways without loss of signaling components. The ballistic delivery of calcium indicators thus opens up many new avenues for further exploration of the structure and function of the nervous system from single spines to neuronal networks.

Original languageEnglish
Pages (from-to)37-43
Number of pages7
JournalJournal of Neuroscience Methods
Volume119
Issue number1
DOIs
StatePublished - 15 Sep 2002
Externally publishedYes

Keywords

  • Ballistic
  • Calcium imaging
  • Calcium indicators
  • Gene gun
  • In vivo
  • Optical imaging

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