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Int J Mol Sci. 2017 Jul 7;18(7). pii: E1461. doi: 10.3390/ijms18071461.

Transgenic Strategies for Sparse but Strong Expression of Genetically Encoded Voltage and Calcium Indicators.

Author information

1
Laboratory for Neuronal Circuit Dynamics, Imperial College London, London W12 0NN, UK. song@knopfel-lab.net.
2
Laboratory for Neuronal Circuit Dynamics, Imperial College London, London W12 0NN, UK. quyen.do-bao14@imperial.ac.uk.
3
Institute for Systems Genomics, Stem Cell Institute, UConn Health, Farmington, CT 06030-3401, USA. antic@uchc.edu.
4
Laboratory for Neuronal Circuit Dynamics, Imperial College London, London W12 0NN, UK. tknopfel@knopfel-lab.net.
5
Centre for Neurotechnology, Institute of Biomedical Engineering, Imperial College London, London SW7 2AZ, UK. tknopfel@knopfel-lab.net.

Abstract

Rapidly progressing development of optogenetic tools, particularly genetically encoded optical indicators, enables monitoring activities of neuronal circuits of identified cell populations in longitudinal in vivo studies. Recently developed advanced transgenic approaches achieve high levels of indicator expression. However, targeting non-sparse cell populations leads to dense expression patterns such that optical signals from neuronal processes cannot be allocated to individual neurons. This issue is particularly pertinent for the use of genetically encoded voltage indicators whose membrane-delimited signals arise largely from the neuropil where dendritic and axonal membranes of many cells intermingle. Here we address this need for sparse but strong expression of genetically encoded optical indicators using a titratable recombination-activated transgene transcription to achieve a Golgi staining-type indicator expression pattern in vivo. Using different transgenic strategies, we also illustrate that co-expression of genetically encoded voltage and calcium indicators can be achieved in vivo for studying neuronal circuit input-output relationships.

KEYWORDS:

controlled recombination; genetically encoded; inducible expression; intersectional; transgenic; voltage indicator

PMID:
28686207
PMCID:
PMC5535952
DOI:
10.3390/ijms18071461
[Indexed for MEDLINE]
Free PMC Article

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