The backstage pass to study
your favorite TRP channel
Access All Areas using
Port-A-Patch, Patchliner or SyncroPatch 384PE
Tom A. Goetze, Mohamed Kreir, Markus Rapedius, Alison Obergrussberger, Nadine Becker, Ilka Rinke, Andrea Brüggemann,
Michael George, Niels Fertig
Nanion Technologies GmbH, Ganghoferstraße 70a, Munich, Germany, info@nanion.deAbstract
TRP channels exhibit a large variety of functional
properties and play diverse cellular and physiologicalroles. Due to their presence in the plasma membrane ofcells from various tissues and even organelles, newbiophysical properties and mechanisms of regulation
are constantly discovered by academic laboratories
over the years. Also, because of their relevance linked
to disease and pain, TRP channels are important targets
in the pharmaceutical industry.
Extensive functional studies of ion channels are often
compromised by technical limitations of theelectrophysiological assay. Nanion’s Port-A-Patch,
Patchliner and SyncroPatch 384PE are specificallydesigned to marry the required experimental flexibility
with high-throughput and Giga-seal data quality.
In order to demonstrate reliable pharmacology
combined with high-throughput electrophysiology weshow data of TRPA1 activated by the agonist
Supercinnamaldehyde and blocked by A-967079 in aconcentration-dependent
manneracrossaTRPV1 and TRPV3 – Patchliner

SyncroPatch 384 chip in a single experiment.
With the aim to highlight experimental flexibility we
present data using the internal perfusion of theSyncroPatch 384PE applying NMM (biotinylated NMethyl Maleimide) and AITC (allyl-iso-thiocyanat) to theintracellular side of the channel.
Intracellular organelles have been accessed using thePort-A-Patch in a report from Chen et al. (NatureComm., 2014). Here, they measured whole lysosomes
and demonstrated how synthetic small-molecule
activators can restore mutant TRPML1 channel function
to rescue disease associated abnormalities inmucolipidosis type IV. Furthermore, single channel
traces of TRPA1 reconstituted into liposomes wererecorded at various cold temperatures and the open
probability was analysed.
Finally, we show data featuring the temperature controlof the Patchliner (available for the SyncroPatch 384PE
shortly) by exposing TRPV1 and -V3 to a range of welldefinedtemperaturesandgenerating
thecorresponding activation curves.

Planar Patch Clamp - Chip replaces pipette

Features:• Parallel 4-8 channel amplifier• Internal perfusion• Temperature control

TRPV1

EC50 = 172 nM ± 54 (n = 4)

T50 = 52°C ± 1.6 (n=4)

Reproducibility

Capsaicin

TRPV3

EC50 = 61 µM ± 8 (n = 11)

T50 = 46°C ± 1.6 (n=6)

52°C

S. Stoelzle, A. Obergrussberger, A. Brüggemann, C.
Haarmann, M. George, R. Kettenhofen , N. Fertig:
Front Pharmacol. 2011; Vol 2: Article 76

J. Behrends, N. Fertig: Neuromethods
2007, Patch-Clamp Analysis, Vol. 38,411-433

TRPA1 – SyncroPatch 384PE
Features:• Modular approach• Parallel 384 amplifier and pipetting head
• Internal perfusion• Temperature control (cell hotel, wholedevice)• Nanion Software (for recording, analysis,batch export)

TRPA1 and TRPML1 – Port-a-Patch
Features:• Experimental
flexibility• Single channel• Whole organelle• Whole cell

Add-ons:• Internal perfusion• External perfusion• Temperature control• Fluorescence slide

TRPA1TRPA1•

whole cell HEK 293
recording• single channel recording (at +60 mV)
purified and reconstituted into GUVs

Recording in PatchControl Software•••

activated by SCMA
blocked by A-967079
concentration range
across the 384 chip

+ 5 µM SCMA+ 15 nM A96707+ 5 µM A96707

L. Moparthia, S. Surverya, M. Kreir, C. Simonsend, P. Kjellboma, E.D.
Högestättd, U. Johansona, P.M. Zygmuntd: PNAS, 2014, Nov
111;47, 16901–16906

Analysis in DataControl Software

Example: Internal Perfusion
Kv1.3 in Jurkat cells
K+(internal)

K+K+

Cs+(internal)

K+

access to ion
TRPML1 • Patch-clamp
channels in organelles•

Cs+„Whole-lysosome“recordingNMM

efficacy in mutants:
SF-22 > PI(3,5)P2lysosomes (HEK293 cells)
SF-22: 10 µM
PI(3,5)P2: 10 µM (diC8)••••
C.-C. Chen, M. Keller, M. Hess, R. Schiffmann, N. Urban,A. Wolfgardt, M. Schaefer, F. Bracher, M. Biel, C. Wahl-Schott,
C. Grimm: Nat Commun. 2014 Aug 14;5-4681•Define QC parameters duringexperiment
Include or reject wells
Manual or automatic analysis
Configurable export of raw andanalysed dataBatch processing

100 s

AITC

Cs+

100 s