ABSTRACT
A mechanosensitive Na(+) current carried by Na(v)1.5 is present in human intestinal circular smooth muscle and contributes to regulation of intestinal motor function. Expression of this channel in different species is unknown. Our aim was to determine if Na(+) currents and message for the alpha subunit of the Na(+) channel (SCN5A) are found in circular smooth muscle cells of human, dog, pig, mouse and guinea pig jejunum. Currents were recorded using patch clamp techniques. Message for SCN5A was investigated using laser capture microdissection and reverse transcription polymerase chain reaction (RT-PCR). Na(+) currents were identified consistently in human and dog smooth muscle cells; however, Na(+) current was not found in pig (0/20) or guinea pig smooth muscle cells (0/21) and found only one mouse cell (1/21). SCN5A mRNA was found in circular muscle of human, dog, and mouse, but not in pig or guinea pig, and not in mouse longitudinal or mucosal layers. In summary, SCN5A message is expressed in, and Na(+) current recorded from, circular muscle layer of human and dog but not from pig and guinea pig. These data show that there are species differences in expression of the SCN5A-encoded Na(v)1.5 channel, suggesting species-specific differences in the electrophysiological response to mechanical and depolarizing stimuli.
Subject(s)
Jejunum/physiology , Mechanoreceptors/physiology , Muscle, Smooth/physiology , Sodium Channels/physiology , Animals , Dogs , Electric Capacitance , Guinea Pigs , Humans , Lasers , Mice , Microdissection , NAV1.5 Voltage-Gated Sodium Channel , NAV1.8 Voltage-Gated Sodium Channel , Patch-Clamp Techniques , RNA, Messenger/analysis , Reverse Transcriptase Polymerase Chain Reaction , Sodium/metabolism , Sodium Channels/genetics , Species Specificity , SwineABSTRACT
Mechanotransduction is required for a wide variety of biological functions. The aim of this study was to determine the effect of activation of a mechanosensitive Ca(2+) channel, present in human jejunal circular smooth muscle cells, on whole cell currents and on membrane potential. Currents were recorded using patch-clamp techniques, and perfusion of the bath (10 ml/min, 30 s) was used to mechanoactivate the L-type Ca(2+) channel. Perfusion resulted in activation of L-type Ca(2+) channels and an increase in outward current from 664 +/- 57 to 773 +/- 72 pA at +60 mV. Membrane potential hyperpolarized from -42 +/- 4 to -50 +/- 5 mV. In the presence of nifedipine (10 microM), there was no increase in outward current or change in membrane potential with perfusion. In the presence of charybdotoxin or iberiotoxin, perfusion of the bath did not increase outward current or change membrane potential. A model is proposed in which mechanoactivation of an L-type Ca(2+) channel current in human jejunal circular smooth muscle cells results in increased Ca(2+) entry and cell contraction. Ca(2+) entry activates large-conductance Ca(2+)-activated K(+) channels, resulting in membrane hyperpolarization and relaxation.
Subject(s)
Calcium Channels, L-Type/physiology , Muscle, Smooth/physiology , Calcium Channel Blockers/pharmacology , Calcium Channels, L-Type/drug effects , Cells, Cultured , Charybdotoxin/pharmacology , Humans , Jejunum/physiology , Membrane Potentials , Muscle Contraction/drug effects , Muscle Contraction/physiology , Nifedipine/pharmacology , Patch-Clamp Techniques , Physical Stimulation , Potassium/metabolism , Potassium Channels/drug effects , Potassium Channels/metabolismABSTRACT
BACKGROUND & AIMS: Gastrointestinal smooth muscle strips devoid of enteric nerve cells can contract in response to stretch, suggesting that mechanosensitivity and mechanotransduction can occur at the level of the smooth muscle cell. The aim of this study was to determine whether stretch-activated calcium channels are present in gastrointestinal smooth muscle cells. METHODS: Whole-cell and single-channel calcium currents were measured from human jejunal circular smooth muscle cells in response to increased intracellular pressure, bath perfusion, and membrane stretch. RESULTS: At 10 mm Hg positive pressure, peak calcium current increased from -36 +/- 10 pA to -53 +/- 13 pA. Bath perfusion at 10 mL/min increased calcium current from -97.7 +/- 14 pA to -122 +/- 16 pA. Single-channel open probability increased in response to negative pipette pressure. All increases were blocked by nifedipine. CONCLUSIONS: A stretch-activated, nifedipine-sensitive calcium channel is present in human jejunal circular smooth muscle cells. The channel is activated by both an increase in intracellular pressure and by external shear forces. The presence of a stretch-activated calcium channel in gastrointestinal smooth muscle cells may allow the smooth muscle cells to act directly as mechanotransducers and to participate in the regulation of smooth muscle tone and intestinal motility.