• Neonatal Medicine
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• 제15권2호
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• pp.196-199
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• 2008

저자들은 Hirschsprung 병을 의심한 환아에서 Cajal세포 감소에 의한 미숙아 가성 장 폐쇄를 경험하였기에 보고하는 바이다. 심한 복부 팽만과 장 폐쇄 증상으로 결장 절개술이 필요한 미숙아에서 ganglion cell의 존재 유무 뿐만 아니라 Cajal 세포의 결핍도 확인할 필요가 있을 것으로 생각된다.

• 대한약침학회지
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• 제16권1호
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• pp.43-49
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• 2013

Objective: Pyungwi-san (PWS) plays a role in a number of physiologic and pharmacologic functions in many organs. Interstitial cells of Cajal (ICCs) are pacemaker cells that generate slow waves in the gastrointestinal (GI) tract. We aimed to investigate the beneficial effects of PWS in mouse small-intestinal ICCs. Methods: Enzymatic digestion was used to dissociate ICCs from the small intestine of a mouse. The whole-cell patch-clamp configuration was used to record membrane potentials from the cultured ICCs. Results: ICCs generated pacemaker potentials in the GI tract. PWS produced membrane depolarization in the current clamp mode. Pretreatment with a $Ca^{2+}$-free solution and a thapsigargin, a $Ca^{2+}$-ATPase, inhibitor in the endoplasmic reticulum, eliminated the generation of pacemaker potentials. However, only when the thapsigargin was applied in a bath solution, the membrane depolarization was not produced by PWS. Furthermore, the membrane depolarizations due to PWS were inhibited not by U-73122, an active phospholipase C inhibitor, but by chelerythrine and calphostin C, protein kinase C inhibitors. Conclusions: These results suggest that PWS might affect GI motility by modulating the pacemaker activity in the ICCs.

• 대한약침학회지
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• 제16권4호
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• pp.36-42
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• 2013

Objectives: Shengmaisan (SMS) is a traditional Chinese medicine prescription widely used for the treatment of diverse organs in Korea. The interstitial cells of Cajal (ICCs) are pacemaker cells that play an important role in the generation of coordinated gastrointestinal (GI) motility. We have aimed to investigate the effects of SMS in the ICCs in the mouse small intestine. Methods: To dissociate the ICCs, we used enzymatic digestions from the small intestine in a mouse. After that, the ICCs were identified immunologically by using the anti-c-kit antibody. In the ICCs, the electrophysiological whole-cell patch-clamp configuration was used to record pacemaker potentials in the cultured ICCs. Results: The ICCs generated pacemaker potentials in the mouse small intestine. SMS produced membrane depolarization with concentration-dependent manners in the current clamp mode. Pretreatment with a $Ca^{2+}$ free solution and thapsigargin, a $Ca^{2+}$-ATPase inhibitor in the endoplasmic reticulum, stopped the generation of the pacemaker potentials. In the case of $Ca^{2+}$-free solutions, SMS induced membrane depolarizations. However, when thapsigargin in a bath solution was applied, the membrane depolarization was not produced by SMS. The membrane depolarizations produced by SMS were inhibited by U-73122, an active phospholipase C (PLC) inhibitors. Furthermore, chelerythrine and calphostin C, a protein kinase C (PKC) inhibitors had no effects on SMS-induced membrane depolarizations. Conclusions: These results suggest that SMS might affect GI motility by modulating the pacemaker activity through an internal $Ca^{2+}$- and PLC-dependent and PKC-independent pathway in the ICCs.

• The Korean Journal of Physiology and Pharmacology
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• 제10권1호
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• pp.7-11
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• 2006

Interstitial cells of Cajal (ICCs) are pacemakers in gastrointestinal tracts, regulating rhythmicity by activating nonselective cation channels (NSCCs). In the present study, we investigated the general characteristics and pH-mediated regulation of pacemaker activity in cultured interstitial cells of Cajal. Under voltage clamp mode and at the holding potential of -60 mV, the I-V relationships and difference current showed that there was no reversal potential and voltage-independent inward current. Also, when the holding potentials were changed from +20 mV to -80 mV with intervals of 20 mV, there was little difference in inward current. In pacemaker activity, the resting membrane potential (RMP) was depolarized (In pH 5.5, $23{\pm}1.5$ mV depolarized) and the amplitude was decreased by a decrease of the extracellular pH. However, in case of increase of extracellular pH, the RMP was slightly hyperpolarized and the amplitude was decreased a little. The melastatin type transient receptor potential (TRPM) channel 7 has been suggested to be required for intestinal pacemaking activity. TRPM7 produced large outward currents and small inward currents by voltage ramps, ranging from +100 to -100 mV from a holding potential of -60 mV. The inward current of TRPM7 was dramatically increased by a decrease in the extracellular pH. At pH 4.0, the average inward current amplitude measured at -100 mV was increased by about 7 fold, compared with the current amplitude at pH 7.4. Changes in the outward current (measured at +100 mV) were much smaller than those of the inward current. These results indicate that the resting membrane potential of pacemaking activity might be depolarized by external acidic pH through TRPM7 that is required for intestinal pacemaking activity.

• 동의생리병리학회지
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• 제25권4호
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• pp.603-607
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• 2011

The purpose of this study is to investigate the effects of Carthami Flos on interstitial cells of Cajal in the gastrointestinal tract. Many regions of the tunica muscularis of the gastrointestinal (GI) tract display spontaneous contraction. These spontaneous contractions are mediated by periodic generation of electrical slow waves. Recent studies have shown that the interstitial cells of Cajal (ICCs) act as pacemakers and conductors of electrical slow waves in gastrointestinal smooth muscles. We investigated the cytotoxicity activity, antioxidant activity, and pacemaking activity. The cytotoxicity activity was measured by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. Antioxidant activities were determined by DPPH (1.1-diphenyl-2-picrylhydrazyl) radical scavenging capacity assay and DCFH-DA (2,7-dichlorofluorescein diacetate) method. The effects of Carthami Flos on the pacemaker potentials in cultured ICCs from murine small intestine were investigated by using whole-cell patch-clamp techniques at $30^{\circ}C$. The addition of Carthami Flos (5, 10, $30{\mu}g$/ml) depolarized the resting membrane potentials in a concentration dependent manner. These results suggest that the GI tract can be targets for Carthami Flos, and their interaction can affect intestinal motility.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제14권2호
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• pp.171-180
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• 2011

목 적: 고지방식이를 섭취한 흰쥐에서 식이 유발인자인 NPY와 장내 운동 조절인자인 Cajal 간질세포의 변화를 관찰하고자 하였다. 대상 및 방법: 동일 연령의 Sprague-Dawley계 수컷 흰쥐를 임의로 대조군(7마리)과 고지방식이군(7마리)으로 분리한 후, 대조군에는 일반 흰쥐 사료를 충분히 섭취시켰으며, 고지방식이군에게는 60% kcal 지방이 포함된 사료(중앙실험동물센터)를 6주 동안 물과 함께 충분히 섭취하도록 하였다. 6주 후 시상하부 조직 절편에 rabbit polyclonal NPY (1 : 1000, Abcam)항체를 반응시키고, 소장의 조직 절편에 rabbit polyclonal c-kit (1 : 400, Neuromics)항체를 반응시켜서 면역조직화학 검사를 시행하였다. 결 과: 고지방식이군의 시상하부에서 NPY의 면역 반응성이 강하게 나타났으며, 특히 PVN에서 NPY의 면역 반응 세포수의 증가를 관찰할 수 있었다. 고지방식이군에서 소장의 Auerbach's plexus 부위의 Cajal 간질 세포의 면역 반응성이 떨어졌으며, 환상근과 종주근 내 Cajal 간질 세포 수도 현저히 감소하였다. 결 론: 고지방식이를 섭취한 흰쥐에서 시상하부 NPY 면역반응 신경세포수와 발현의 증가는 NPY의 저항성과 관련되며, 소장에서 Cajal 간질 세포의 감소는 장운동의 이상을 초래할 수 있음을 나타낸다.

• 대한의생명과학회지
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• 제18권3호
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• pp.218-226
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• 2012

Dopamine is an enteric neurotransmitter that regulates gastrointestinal motility. This study was done to investigate whether dopamine modulates spontaneous pacemaker activity in cultured interstitial cells of Cajal (ICCs) from mouse using whole cell patch clamp technique, RT-PCR and live $Ca^{2+}$ imaging analysis. ICCs generate pacemaker inward currents at a holding potential of -70 mV and generate pacemaker potentials in current-clamp mode. Dopamine did not change the frequency and amplitude of pacemaker activity in small intestinal ICCs. On the contrary dopamine reduced the frequency and amplitude of pacemaker activity in large intestinal ICCs. RT-PCR analysis revealed that Dopamine2 and 4-receptors are expressed in c-Kit positive ICCs. Dopamine2 and 4 receptor agonists inhibited pacemaker activity in large intestinal ICCs mimicked those of dopamine. Domperidone, dopamine2 receptor antagonist, increased the frequency of pacemaker activity of large intestinal ICCs. In $Ca^{2+}$-imaging, dopamine inhibited spontaneous intracellular $Ca^{2+}$ oscillations of ICCs. These results suggest that dopamine can regulate gastrointestinal motility through modulating pacemaker activity of large intestinal ICCs and dopamine effects on ICCs are mediated by dopamine2 receptor and intracellular $Ca^{2+}$ modulation.

• 동의생리병리학회지
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• 제27권1호
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• pp.112-117
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• 2013

We studied the modulation of pacemaker activities by Samchulkunbi-tang (SCKB) in cultured interstitial cells of Cajal (ICC) from murine small intestine with the whole-cell patch-clamp technique. Externally applied SCKB produced membrane depolarization in the current-clamp mode. The pretreatment with $Ca^{2+}$-free solution and thapsigargin, a $Ca^{2+}$-ATPase inhibitor in endoplasmic reticulum, abolished the generation of pacemaker potentials and suppressed the SCKB-induced action. The application of flufenamic acid (a nonselective cation channel blocker) abolished the generation of pacemaker potentials by SCKB. However, the application of niflumic acid (a chloride channel blocker) did not inhibit the generation of pacemaker potentials by SCKB. In addition, the membrane depolarizations were inhibited by not only GDP-${\beta}$-S, which permanently binds G-binding proteins, but also U-73122, an active phospholipase C inhibitor. These results suggest that SCKB modulates the pacemaker activities by nonselective cation channels and external $Ca^{2+}$ influx and internal $Ca^{2+}$ release via G-protein and phospholipase C-dependent mechanism. Therefore, the ICC are targets for SCKB and their interaction can affect intestinal motility.

• The Korean Journal of Physiology and Pharmacology
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• 제10권4호
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• pp.193-198
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• 2006

Many gastrointestinal muscles show electrical oscillation, so-called 'slow wave', originated from interstitial cells of Cajal (ICCs). Thus, a technique to freshly isolate the cells is indispensable to explore the electrophysiological properties of the ICCs. To apply an enzyme solution on the serosal surface for cell isolation, the intestine was inverted and 0.02% trypsin solution and 0.04% collagenase solution were applied to serosal cavity. After the enzyme treatment, mucosal layer was removed and longitudinal muscle layer was gently separated from the rest of tissue. The thin layer was stretched in the recording chamber and mounted on an inverted microscope. Using ${\beta}-escine$, perforated whole cell patch clamp technique was used. Under a microscope, the tissue showed smooth muscle cells and interstitial cells around the myenteric plexus. Under voltage clamp condition, three types of membrane potential were recorded. One group of interstitial cells, which were positive to methylene blue and CD34, showed spontaneous outward current. These cells had bipolar shape and were considered as fibroblast-like cells because of their peculiar shape and arrangement. Another group, positive to c-kit and methylene blue, showed spontaneous inward current. These cells had more rounded shape and processes and were considered as ICCs. The third, positive to c-kit and had granules containing methylene blue, showed quiet membrane potentials under the voltage-clamp mode. These cells appeared to be resident macrophages. Therefore, in the freshly isolated thin tissue preparation, methylene blue could easily identify three types of cells rather than morphological properties. Using this method, we were able to study electrical properties of fibroblast and residential macrophage as well as myenteric ICCs.

• The Korean Journal of Physiology and Pharmacology
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• 제12권1호
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• pp.25-30
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• 2008

Although many studies show that thromboxane $A_2\;(TXA_2)$ has the action of gastrointestinal (GI) motility using GI muscle cells and tissue, there are no reports on the effects of $TXA_2$ on interstitial cells of Cajal (ICC) that function as pacemaker cells in GI tract. So, we studied the modulation of pacemaker activities by $TXA_2$ in ICC with whole cell patch-clamp technique. Externally applied $TXA_2\;(5{\mu}M)$ produced membrane depolarization in current-clamp mode and increased tonic inward pacemaker currents in voltage-clamp mode. The tonic inward currents by $TXA_2$ were inhibited by intracellular application of GDP-${\beta}$-S. The pretreatment of ICC with $Ca^{2+}$ free solution and thapsigargin, a $Ca^{2+}$-ATPase inhibitor in endoplasmic reticulum, abolished the generation of pacemaker currents and suppressed the $TXA_2$-induced tonic inward currents. However, chelerythrine or calphostin C, protein kinase C inhibitors, did not block the $TXA_2$-induced effects on pacemaker currents. These results suggest that $TXA_2$ can regulate intestinal motility through the modulation of ICC pacemaker activities. This modulation of pacemaker activities by $TXA_2$ may occur by the activation of G protein and PKC independent pathway via extra and intracellular $Ca^{2+}$ modulation.