127
Original Paper
Cell Physiol Biochem 2005;16:127-132 Accepted: April 18, 2005 Cellular Physiology Cellular Physiology Cellular Physiology Cellular Physiology Cellular Physiology
and Biochemistr and Biochemistr and Biochemistr and Biochemistr and Biochemistry
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Establishment and Characterization of a Mouse
Embryonic Heart Slice Preparation
Frank Pillekamp, M.D.
1,2,a
, Michael Reppel, M.D.
2,a
, Vera Dinkelacker,
M.D.
3,4
, Yaqi Duan
2
, Nathalie Jazmati
2
, Wilhelm Bloch, M.D.
5
, Konrad
Brockmeier, M.D.
1
, Juergen Hescheler, M.D.
2
, Bernd K.
Fleischmann, M.D.
3
and Ruediger Koehling, M.D.
4
1
Pediatric Cardiology and
2
Institute of Neurophysiology, University of Cologne,
3
Institute of Physiology I
and
4
Department of Epileptology, University of Bonn,
5
Department of Molecular and Cellular Sport Medicine,
German Sport University, Cologne,
a
authors contributed equally to the manuscript
Bernd K. Fleischmann, M.D.
Institute of Physiology I, Live & Brain Center, University of Bonn
Sigmund-Freud-Str. 25, D-53105 Bonn (Germany)
Tel. +49-228-6885-200, Fax +49-228-6885-201
E-Mail bernd.fleischmann@uni-bonn.de
Key Words
Myocytes, cardiac/*physiology Action potentials
Electrophysiology/*methods Electrophysiology/
*embryology Heart/*physiology Animal models
Abstract
Background: In contrast to isolated cells, the anatomic
and functional integrity of tissue slices remains
preserved. Aim of the study was to establish the slice
technique in embryonic mouse hearts in order to
perform physiological and pharmacological
investigations of wild-type mice and genetically
engineered mouse models of heart disease. Methods:
Ventricular slices (thickness: 300 µm) were cut from
agar-embedded embryonic mouse hearts (ED 16.5-
18.5) with a vibratome. Histology, immunostaining with
markers for apoptosis induction, intracellular
recordings with sharp electrodes and field potential
recordings using microelectrode arrays were
performed to assess viability. Results: Slices exhibited
normal histology without prominent signs of apoptosis
for at least 24 hours. Intracellular recordings revealed
the typical electrophysiological fingerprint of
ventricular cardiomyocytes. Field potential recordings
proved that adrenergic and muscarinic signaling was
preserved. Conclusion: Functionally intact heart slices
can be generated from murine embryos.
Introduction
The advent of the brain slice preparation led to a
dramatic improvement of the understanding of cerebral
activity because it has enabled studies of cellular function
in the context of an intact tissue [1]. This is particularly
important for structures like the brain and the heart in
which intercellular cross-talk is critical for complex organ
function. Furthermore, aggressive dissociation methods
required to prepare isolated cells, often have adverse
effects on the function of ion channels and receptors.
Thus, we sought to establish an intact heart slice
preparation so that we might better understand
mechanisms of cardiac physiology in the future. To date,
relatively little progress has been made in establishing a
viable heart slice preparation [2, 3] and in recording
physiological activity of cardiac slices [4]. This is in part
due to the fact that the heart contains a significant amount