Biomedical subjects

Jan D Huizinga

Publications and source records attributed to Jan D Huizinga.

5 recordsLinked to original sources

PubMed2003

ERG K+ currents regulate pacemaker activity in ICC.

Ether-à-go-go-related gene (ERG) K channels have been implicated in the generation of pacemaker activities in the heart. To study the presence and function of ERG K channels in the pacemaker cells of the small intestine [the interstitial cells of Cajal (ICC)], a combination of patch-clamp techniques, tissue and live cell immunohistochemistry, RT-PCR, and in vitro functional studies were performed. Nonenzymatically isolated ICC in culture were identified by vital staining and presence of rhythmic inward currents. RT-PCR showed the presence of ERG mRNA in the intestinal musculature, and immunohistochemistry on tissue and cultured cells demonstrated that protein similar to human ERG was concentrated on ICC in the Auerbach's plexus region. Whole cell ERG K+ currents were evoked on hyperpolarization from 0 mV (but not from -70 mV) up to -120 mV and showed strong inward rectification. The currents were inhibited by E-4031, cisapride, La3+, and Gd3+ but not by 50 microM Ba2+. The ERG K+ inward current had a typical transient component with fast activation and inactivation kinetics followed by significant steady-state current. E-4031 also inhibited tetraethylammonium (TEA)-insensitive outward current indicating that the ERG K+ current is operating at depolarizing potentials. In contrast to TEA, blockers of the ERG K+ currents caused marked increase in tissue excitability as reflected by an increase in slow-wave duration and an increase in superimposed action potential activity. In summary, ERG K channels in ICC contribute to the membrane potential and play a role in regulation of pacemaker activity of the small intestine.

Animals↗

PubMed2002

Heterogeneous expression of transient outward currents in smooth muscle cells of the mouse small intestine.

The objective for this paper was to characterize the transient outward current (I(to)) present in smooth muscle cells of the intestinal external muscularis. Two populations of cells were identified, one with a fast rate of I(to) inactivation (tau < 100 ms) and another with a slow rate of I(to) inactivating (tau > 150 ms). The chord conductance for the fast I(to) was smaller than the chord conductance of the slow I(to) (0.5 +/- 0.1 vs. 1.3 +/- 0.1 nS pF(-1), respectively). The inactivation was fitted by mono-exponentials to give a tau for the fast and slow I(to) of 44 and 229 ms, respectively. Combined plots of voltage dependent activation and inactivation processes revealed voltage ranges where window currents were possible; a 20 mV range for the fast I(to) from -56 to -36 mV and a 47 mV range for the slow I(to) from -42 to +5 mV. The fast I(to) recovered more quickly from inactivation than the slow I(to); tau(fast I(to)) = 11 +/- 4 ms compared to tau(slow I(to)) = 42 +/- 16 ms. The effect of different rates of depolarization on I(to) activation was examined. The plots of peak currents evoked by different rates of depolarization were well fitted by inverse exponential functions. The fast I(to) had a larger response to fast rates of depolarization by having a tau of 2 +/- 1 mV ms(-1) with maximal activation (95 % complete) at 5 mV ms(-1). The slow I(to) had a tau of 14 +/- 9 mV ms(-1) with maximal activation (95 % complete) at 42 mV ms(-1). The properties of these currents suggest that the two transient outward currents may contribute differently to slow waves and action potentials generated by the smooth muscle cells.

Animals↗

PubMed2002

Pathology of interstitial cells of Cajal in relation to inflammation revealed by ultrastructure but not immunohistochemistry.

The role of interstitial cells of Cajal associated with Auerbach's plexus (ICC-AP) in the pathophysiology of inflammation-induced abnormalities in gut motor activity is poorly understood. Therefore we applied a well-described model of inflammation (infection by Trichinella spiralis) to the mouse small intestine where the structure and function of ICC-AP are best known. Electron microscopic evaluation revealed that 1 to 3 days after infection, selective and patchy damage to the ICC processes occurred, thereby disrupting contacts between these ICC and smooth muscle cells as well as ICC and nerves, which was associated with disordered electrical activity and abnormal peristalsis. Ten to 15 days after infection, damage to ICC-AP was maximal and now involving the cell body and major processes. Marked synthetic activity and regrowth of their processes occurred from day 3 onward and recovery was completed at day 40 after infection. No changes to the network of ICC-AP were seen with c-Kit immunohistochemistry. From day 1 after infection, macrophages infiltrated the AP area, making close contact including peg-and-socket-like junctions with smooth muscle cells and ICC-AP but up to day 6 after infection without any sign of phagocytosis. By day 6 after infection, lymphocytes entered the musculature forming close contacts with ICC-AP. This was not associated with damage to ICC-AP but with proliferation of rough endoplasmic reticulum. From day 23 onward, immune cells withdrew from the musculature except macrophages, resulting in a markedly increased population of macrophages in the AP area at day 60 after infection.

Animals↗

PubMed2002

High-conductance chloride channels generate pacemaker currents in interstitial cells of Cajal.

BACKGROUND & AIMS: Interstitial cells of Cajal (ICCs) are responsible for slow, wave-driven, rhythmic, peristaltic motor patterns in the gastrointestinal tract. The aim was to identify and characterize the ion channels that generate the underlying pacemaker activity. METHODS: Single ion channel recordings were obtained from nonenzymatically isolated ICCs and studied by using the cell attached and inside-out configurations of the patch clamp technique. RESULTS: A high-conductance chloride channel was observed in ICCs that was spontaneously and rhythmically active at the same frequency as the rhythmic inward currents defining ICC pacemaker activity, 20-30 cycles/min at room temperature. Main conductance levels occurred between 122-144 pS and between 185-216 pS. Periodicity in the channel opening coincided with periodicity in membrane potential change, hence, at the single channel level, chloride channels were seen to be associated with the generation of rhythmic changes in membrane potential. CONCLUSIONS: ICCs harbor high-conductance chloride channels that participate in the generation of pacemaker activity and may become a target for pharmacologic treatment of gut motor disorders.

Animals↗

PubMed2002

Gastric electrical stimulation in intractable symptomatic gastroparesis.

BACKGROUND: The treatment of gastroparesis remains unsatisfactory despite prokinetic and anti-emetic drugs. Gastric electrical stimulation has been proposed as a therapeutic option. We have assessed the effect of gastric electrical stimulation on symptoms, medical treatment, body weight and gastric emptying in patients with intractable symptomatic gastroparesis in a non-placebo-controlled study. METHODS: In this multicenter study, 38 highly symptomatic patients with drug-refractory gastroparesis were enrolled. Patients first received temporary electrical stimulation using percutaneous electrodes. The 33 responders to temporary stimulation then underwent surgical implantation of a permanent stimulator. Severity of vomiting and nausea was assessed before and after stimulation. Patients were reassessed 3, 6, and 12 months after permanent implantation. RESULTS: With stimulation, 35/38 patients (97%) experienced >80% reduction in vomiting and nausea. This effect persisted throughout the observation period (2.9-15.6 months, 341 patient-months). Gastric emptying did not initially change, but improved in most patients at 12 months. At 1 year, the average weight gain was 5.5% and 9/14 patients initially receiving enteral or parenteral nutrition were able to discontinue it. CONCLUSION: Electrical stimulation of the stomach has an immediate and potent anti-emetic effect. It offers a safe and effective alternative for patients with intractable symptomatic gastroparesis.

Adolescent↗