Advanced Methods in Cellular Electrophysiology
Advanced Methods in Cellular Electrophysiology
Research Topics uitklapper, klik om te openen
An important observation is that cells of an organ often replicate the primary function of the organ in which they reside; neurons integrate signals from their surroundings and heart muscle cells (cardiomyocytes) contract in response to an electrical stimulus. In our department we aim to study the cellular physiology of cardiomyocytes using conventional approaches that rely on isolated, single cardiomyocytes (such as patch clamp electrophysiology), but have also started work on novel methods that will help to increase our insight in the function of cardiomyocytes within the heart. For this we use optogenetic sensors that allow us to record e.g. intracellular calcium concentration or membrane potential of cardiomyocytes within the heart.
Another important aspect of our work is the development of real-time simulations of ion channels, the so-called dynamic clamp technique. The most important application is to simulate cardiac IK1 channels that are missing from stem cell-derived cardiomyocytes (iPSC-CM). By combining the real-time simulated ion channels with human iPSC-CM, we can create a hybrid human cardiomyocyte model that is more similar to human adult cardiomyocytes. The main benefit of this approach is that it enables more specific in vitro testing of drugs that might negatively affect the human heart rhythm. Current focus is to enhance the throughput of this approach, which resulted in the establishment of a fully automated dynamic clamping system which is integrated with an automated patch clamping device (Patchliner) developed by Nanion Technologies. Nanion has obtained a licence for our technology, and has brought the system to the market under the name Dynamite8.
Key Publications uitklapper, klik om te openen
Lei CL, Fabbri A, Whittaker DG, Clerx M, Windley MJ, Hill AP, Mirams GR, de Boer TP. A nonlinear and time-dependent leak current in the presence of calcium fluoride patch-clamp seal enhancer. Wellcome Open Res. 2021 Nov 2;5:152. doi: 10.12688/wellcomeopenres.15968.2. PMID: 34805549; PMCID: PMC8591515.
Koopman CD, De Angelis J, Iyer SP, Verkerk AO, Da Silva J, Berecki G, Jeanes A, Baillie GJ, Paterson S, Uribe V, Ehrlich OV, Robinson SD, Garric L, Petrou S, Simons C, Vetter I, Hogan BM, de Boer TP, Bakkers J, Smith KA. The zebrafish grime mutant uncovers an evolutionarily conserved role for Tmem161b in the control of cardiac rhythm. Proc Natl Acad Sci U S A. 2021 Mar 2;118(9):e2018220118. doi: 10.1073/pnas.2018220118. PMID: 33597309; PMCID: PMC7936323.
Fabbri A, Goversen B, Vos MA, van Veen TAB, de Boer TP. Required GK1 to Suppress Automaticity of iPSC-CMs Depends Strongly on IK1 Model Structure. Biophys J. 2019 Dec 17;117(12):2303-2315. doi: 10.1016/j.bpj.2019.08.040. Epub 2019 Sep 13. PMID: 31623886; PMCID: PMC6990378.
Goversen B, Becker N, Stoelzle-Feix S, Obergrussberger A, Vos MA, van Veen TAB, Fertig N, de Boer TP. A Hybrid Model for Safety Pharmacology on an Automated Patch Clamp Platform: Using Dynamic Clamp to Join iPSC-Derived Cardiomyocytes and Simulations of Ik1 Ion Channels in Real-Time. Front Physiol. 2018 Jan 19;8:1094. doi: 10.3389/fphys.2017.01094. PMID: 29403387; PMCID: PMC5782795.
van Opbergen CJM, Koopman CD, Kok BJM, Knöpfel T, Renninger SL, Orger MB, Vos MA, van Veen TAB, Bakkers J, de Boer TP. Optogenetic sensors in the zebrafish heart: a novel in vivo electrophysiological tool to study cardiac arrhythmogenesis. Theranostics. 2018 Sep 9;8(17):4750-4764. doi: 10.7150/thno.26108. PMID: 30279735; PMCID: PMC6160779.
Facilities Involved uitklapper, klik om te openen
People Involved uitklapper, klik om te openen
Contact uitklapper, klik om te openen
Teun de Boer