Single mechano-gated channels activated by mechanical deformation of acutely isolated cardiac fibroblasts from rats.
OBJECTIVE
Mechanosensitive conductances were reported in cardiac fibroblasts, but the properties of single channels mediating their mechanosensitivity remain uncharacterized. The aim of this work was to investigate single mechano-gated channels (MGCs) activated by mechanical deformations of cardiac fibroblasts.


METHODS
Currents through single MGCs and mechanosensitive whole-cell currents were recorded from isolated rat atrial fibroblasts using the cell-attached and whole-cell patch-clamp configurations respectively. Defined mechanical stress was applied via the patch pipette used for the whole-cell recordings.


RESULTS
Under resting conditions occasional short openings of two types of single MGCs with conductances of 43 and 87 pS were observed. Both types of channels displayed a linear current-voltage relationship with the reversal potential around 0 mV. Small (1 microm) mechanical deformations affected neither single nor whole-cell mechano-gated currents. Cell compressions (2, 3 and 4 microm) augmented the whole-cell currents and increased the frequency and duration of single channel openings. Cell stretches (2, 3 and 4 microm) inactivated the whole-cell currents and abolished the activity of single MGCs. Gd(3+) (8 microm) blocked the whole-cell currents within 5 min. No single channel activity was observed in the cell-attached mode when Gd(3+) was added to the intrapipette solution. Cytochalasin D and colchicine (100 microm each) completely blocked both the whole-cell and single channel currents.


CONCLUSIONS
These findings show that rat atrial fibroblasts express two types of MGCs whose activity is governed by cell deformation. We conclude that fibroblasts can sense the direction of applied stress and contribute to mechano-electrical coupling in the heart.
Kamkin
A
A
Department of Fundamental and Applied Physiology, Russian State Medical University, Moscow, Russia. kamkin.a@g23.relcom.ru <kamkin.a@g23.relcom.ru>
Kirischuk
S
S
Kiseleva
I
I
D000818:Animals; D002469:Cell Separation; D048430:Cell Shape / Q000502:physiology; D003078:Colchicine / Q000494:pharmacology; D015638:Cytochalasin D / Q000494:pharmacology; D004558:Electric Stimulation; D004594:Electrophysiology; D005347:Fibroblasts / Q000502:physiology* / Q000648:ultrastructure; D006325:Heart Atria / Q000166:cytology; D015640:Ion Channel Gating / Q000502:physiology*; D007473:Ion Channels / Q000502:physiology*; D008297:Male; D032383:Myocytes, Cardiac / Q000502:physiology*; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D011312:Pressure; D051381:Rats; D012996:Solutions
D007473:Ion Channels; D019384:Nucleic Acid Synthesis Inhibitors; D012996:Solutions; D015638:Cytochalasin D; D003078:Colchicine