Abstract
Dynamic clamp is a powerful method that allows the introduction of artificial electrical components into target cells to simulate ionic conductances and synaptic inputs. This method is based on a fast cycle of measuring the membrane potential of a cell, calculating the current of a desired simulated component using an appropriate model and injecting this current into the cell. Here we present a dynamic clamp protocol using free, fully integrated, open-source software (StdpC, for spike timing-dependent plasticity clamp). Use of this protocol does not require specialist hardware, costly commercial software, experience in real-time operating systems or a strong programming background. The software enables the configuration and operation of a wide range of complex and fully automated dynamic clamp experiments through an intuitive and powerful interface with a minimal initial lead time of a few hours. After initial configuration, experimental results can be generated within minutes of establishing cell recording.
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Acknowledgements
T.N. is indebted to R.D. Pinto for the initial development work of Dynclamp 2/4, the early predecessor of StdpC, and to A. Szücs for years of testing StdpC. This work was financed partially by an RCUK fellowship to T.N. The work also received financial support from an MRC research grant held by G.K. and from BBSRC and Wellcome Trust research grants held by K.S.
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Contributions
I.K. set up preparations and designed and conducted the electrophysiological experiments using invertebrate neurons, V.M. set up preparations, designed and conducted electrophysiological experiments using vertebrate neurons and designed figures, M.C. contributed to the electrophysiological experiments using invertebrate neurons, D.S. worked on the software and drafted parts of the manuscript, K.S. designed and supervised vertebrate experimentation and wrote the manuscript, G.K. designed and supervised the experiments using invertebrate neurons and T.N. developed the method and software, wrote the manuscript, produced the supplementary movies, established dynamic clamp at Sussex together with G.K. and coordinated the whole effort. All authors were involved in revising the manuscript.
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Supplementary information
Supplementary Video 1: Introduction to the dynamic clamp hardware and setup.
This short movie introduces the main hardware components used in a dynamic clamp experiment with StdpC. Thomas Nowotny and Michael Crossley introduce the hardware (electrophysiology rig, amplifier, data acquisition, dynamic clamp data acquisition board), the Lymnaea preparation, and an example of a pattern clamp experiment. (MOV 44750 kb)
Supplementary Video 2: StdpC user interface and configuration
This video gives a brief introduction into how to use the StdpC graphical user interface and how to configure StdpC. We cover general configuration, configuring input and output channels and how to configure a typical simulated chemical synapse. (MOV 23826 kb)
Supplementary Equations: Equations describing the K+ conductance introduced in the Anticipated Results (see also Fig. 3 B).
The supplementary equations described in this supplement define a delayed rectifier potassium current following the model of a hippocampal cell given in ref 48. (PDF 17 kb)
Supplementary Methods: Reagents and solutions used for the Anticipated Results.
In this supplement we provide the details on the reagents and the exact solutions used in the Anticipated Results. Solutions are described for Lymnaea saline, hippocampal bath solution, sharp electrode internal solution and patch electrode internal solution. (PDF 79 kb)
Supplementary Fig. 1: Details of EPSPs and EPSCs of the simulated synaptic conductance shown in Fig. 3A.
A) EPSPs for simulated maximal synaptic conductance of 10, 20, 30, 50, 80, 120, 170, 230, 300, 390, 490, and 600 nS aligned to the occurrence of the pre-synaptic spike. For each maximal conductance, 5 individual EPSPs (grey lines) and their average (colored lines) are shown. The other parameters were as in Fig. 3A, i.e. Vrev= 0 mV, tausyn= 10 ms, and Vslope= 25 mV. (PDF 724 kb)
B) Corresponding EPSCs. The colors of A and B are matched to mark the corresponding data sets.
Animal care and use protocols complied with Home Office (UK) guidelines.
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Kemenes, I., Marra, V., Crossley, M. et al. Dynamic clamp with StdpC software. Nat Protoc 6, 405–417 (2011). https://doi.org/10.1038/nprot.2010.200
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DOI: https://doi.org/10.1038/nprot.2010.200
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