Effect of Houpu Sanwu Tang on Small Intestine Interstitial Cells of Cajal and iNOS in Postoperative Ileus Rats / 中国实验方剂学杂志

Objective: To evaluate the effect of Houpu Sanwu Tang on the postoperative ileus (POI), and observe its underlying mechanisms of action on interstitial cells of cajal (ICC) and inducible nitric oxide synthase(iNOS) regulation of POI. Method: Totally 87 healthy adult male SD rats were randomly divided into sham operation group, saline control group and Houpu Sanwu Tang group at low, medium and high doses. Houpu Sanwu Tang low, middle and high dose groups received orally Houpu Sanwu Tang(2.25,4.5,9 g·kg-1); Sham operation group (Sham operation) and saline control group (Saline control) received orally normal saline. Surgical procedure was used to induce the postoperative ileus. Changes in intestinal propulsion rate, intestinal mucosal injury and small intestine expression of c-kit and iNOS among these groups were detected. Result: Intestinal propulsion rate was significantly higher in Houpo Sanwu Tang group than that in Saline control group (PPPPConclusion: Houpu Sanwu Tang can improve the intestinal propulsion rate and the recovery in POI rats. The mechanisms are related to the inhibition of the generation of iNOS, the alleviation of inflammatory response, and increase of the number of ICC, so as to ensure its normal function, and improve the intestinal dynamic disorder.

Effects of Electroacupuncture on Ultrastructure of Interstitial Cells of Cajal and Stem Cell Factor-kit Signal Pathway of Gastric Antrum in Diabetic Gastroparesis Rats / 针刺研究

OBJECTIVE: To observe the effects of electroacupuncture (EA) on gastrointestinal motility and the ultrastructure of interstitial cells of Cajal (ICC) and the expressions of c-kit receptor protein and stem cell factor (SCF) mRNA in diabetic gastroparesis (DGP) rats, so as to explore its mechanism. METHODS: Fifty SD rats were randomly divided into normal, model, acupoint, non-acupoint and metoclopramide groups (n＝10 rats/group). DGP model was established by intraperitoneal injection of streptozotocin (STZ, 2%), and raised with high-sugar high-fat diet irregularly. EA (sparse-dense, 10 Hz/50 Hz, 2 mA, 20 min) was applied at "Zusanli" (ST 36), "Liangmen" (ST 21) and "Sanyinjiao" (SP 6), and the corresponding non-acupoints of the 3 acupoints, daily for 15 days. The rats in metoclopramide group received intragastric administration of metoclopramide (1.7%, 1 mL/100 g) for 15 days, once a day. Blood sugar was determined with One Touch blood glucose test paper. The gastric emptying rate (GER) and the intestinal propulsion rate (IPR) were measured by intragastric phenol red. The ultrastructure of ICC was detected by transmission electron microscopy. The expression levels of c-kit receptor protein and SCF mRNA of gastric antrum were examined respectively by Western blot and RT-PCR. RESULTS: Compared with the normal group, the blood glucose significantly increased in the model group (P<0.01), while the GER, IRP and the expression level of SCF mRNA in the gastric antrum significantly decreased (P<0.01), and the ultrastructure of ICC appeared apoptosis-like changes. The blood glucose of the EA group was obviously decreased compared with that of the model group (P<0.05); the GER and IRP significantly increased(P<0.05, P<0.01); the expression level of SCF mRNA increased (P<0.01), the number of ICC increased and its ultrastructure was repaired. There was some relief on ICC ultrastructure in the acupoint group compared with that in the non-acupoint group; and SCF mRNA increased (P<0.05). There was no significant difference on c-kit receptor expression among all the modeling groups (P＞0.05). CONCLUSIONS: EA at ST 36, etc. can regulate the blood glucose and improve gastrointestinal emptying in DGP rats. The mechanism may be related to up-regulating SCF mRNA, repairing ICC ultrastructure, restoring the pacing function, and improving gastrointestinal motility.

Pituitary Adenylate Cyclase-activating Polypeptide Inhibits Pacemaker Activity of Colonic Interstitial Cells of Cajal

This study aimed to investigate the effect of pituitary adenylate cyclase-activating peptide (PACAP) on the pacemaker activity of interstitial cells of Cajal (ICC) in mouse colon and to identify the underlying mechanisms of PACAP action. Spontaneous pacemaker activity of colonic ICC and the effects of PACAP were studied using electrophysiological recordings. Exogenously applied PACAP induced hyperpolarization of the cell membrane and inhibited pacemaker frequency in a dose-dependent manner (from 0.1 nM to 100 nM). To investigate cyclic AMP (cAMP) involvement in the effects of PACAP on ICC, SQ-22536 (an inhibitor of adenylate cyclase) and cell-permeable 8-bromo-cAMP were used. SQ-22536 decreased the frequency of pacemaker potentials, and cell-permeable 8-bromo-cAMP increased the frequency of pacemaker potentials. The effects of SQ-22536 on pacemaker potential frequency and membrane hyperpolarization were rescued by co-treatment with glibenclamide (an ATP-sensitive K+ channel blocker). However, neither N(G)-nitro-L-arginine methyl ester (L-NAME, a competitive inhibitor of NO synthase) nor 1H-[1,2,4]oxadiazolo[4,3-alpha]quinoxalin-1-one (ODQ, an inhibitor of guanylate cyclase) had any effect on PACAP-induced activity. In conclusion, this study describes the effects of PACAP on ICC in the mouse colon. PACAP inhibited the pacemaker activity of ICC by acting through ATP-sensitive K+ channels. These results provide evidence of a physiological role for PACAP in regulating gastrointestinal (GI) motility through the modulation of ICC activity.

Interplay of Hydrogen Sulfide and Nitric Oxide on the Pacemaker Activity of Interstitial Cells of Cajal from Mouse Small Intestine / 전남의대학술지

We studied whether nitric oxide (NO) and hydrogen sulfide (H2S) have an interaction on the pacemaker activities of interstitial cells of Cajal (ICC) from the mouse small intestine. The actions of NO and H2S on pacemaker activities were investigated by using the whole-cell patch-clamp technique and intracellular Ca2+ analysis at 30degrees C in cultured mouse ICC. Exogenously applied (+/-)-S-nitroso-N-acetylpenicillamine (SNAP), an NO donor, or sodium hydrogen sulfide (NaHS), a donor of H2S, showed no influence on pacemaker activity (potentials and currents) in ICC at low concentrations (10 microM SNAP and 100 microM NaHS), but SNAP or NaHS completely inhibited pacemaker amplitude and pacemaker frequency with increases in the resting currents in the outward direction at high concentrations (SNAP 100 microM and NaHS 1 mM). Co-treatment with 10 microM SNAP plus 100 microM NaHS also inhibited pacemaker amplitude and pacemaker frequency with increases in the resting currents in the outward direction. ODQ, a guanylate cyclase inhibitor, or glibenclamide, an ATP-sensitive K+ channel inhibitor, blocked the SNAP+NaHS-induced inhibition of pacemaker currents in ICC. Also, we found that SNAP+NaHS inhibited the spontaneous intracellular Ca2+ ([Ca2+]i) oscillations in cultured ICC. In conclusion, this study describes the enhanced inhibitory effects of NO plus H2S on ICC in the mouse small intestine. NO+H2S inhibited the pacemaker activity of ICC by modulating intracellular Ca2+. These results may be evidence of a physiological interaction of NO and H2S in ICC for modulating gastrointestinal motility.

5-Hydroxytryptamine Generates Tonic Inward Currents on Pacemaker Activity of Interstitial Cells of Cajal from Mouse Small Intestine

In this study we determined whether or not 5-hydroxytryptamine (5-HT) has an effect on the pacemaker activities of interstitial cells of Cajal (ICC) from the mouse small intestine. The actions of 5-HT on pacemaker activities were investigated using a whole-cell patch-clamp technique, intracellular Ca2+ ([Ca2+]i) analysis, and RT-PCR in ICC. Exogenously-treated 5-HT showed tonic inward currents on pacemaker currents in ICC under the voltage-clamp mode in a dose-dependent manner. Based on RT-PCR results, we found the existence of 5-HT2B, 3, 4, and 7 receptors in ICC. However, SDZ 205557 (a 5-HT4 receptor antagonist), SB 269970 (a 5-HT7 receptor antagonist), 3-tropanylindole - 3 - carboxylate methiodide (3-TCM; a 5-HT3 antagonist) blocked the 5-HT-induced action on pacemaker activity, but not SB 204741 (a 5-HT2B receptor antagonist). Based on [Ca2+]i analysis, we found that 5-HT increased the intensity of [Ca2+]i. The treatment of PD 98059 or JNK II inhibitor blocked the 5-HT-induced action on pacemaker activity of ICC, but not SB 203580. In summary, these results suggest that 5-HT can modulate pacemaker activity through 5-HT3, 4, and 7 receptors via [Ca2+]i mobilization and regulation of mitogen-activated protein kinases.

Interplay of Hydrogen Sulfide and Nitric Oxide on the Pacemaker Activity of Interstitial Cells of Cajal from Mouse Small Intestine / 전남의대학술지

We studied whether nitric oxide (NO) and hydrogen sulfide (H2S) have an interaction on the pacemaker activities of interstitial cells of Cajal (ICC) from the mouse small intestine. The actions of NO and H2S on pacemaker activities were investigated by using the whole-cell patch-clamp technique and intracellular Ca2+ analysis at 30degrees C in cultured mouse ICC. Exogenously applied (+/-)-S-nitroso-N-acetylpenicillamine (SNAP), an NO donor, or sodium hydrogen sulfide (NaHS), a donor of H2S, showed no influence on pacemaker activity (potentials and currents) in ICC at low concentrations (10 microM SNAP and 100 microM NaHS), but SNAP or NaHS completely inhibited pacemaker amplitude and pacemaker frequency with increases in the resting currents in the outward direction at high concentrations (SNAP 100 microM and NaHS 1 mM). Co-treatment with 10 microM SNAP plus 100 microM NaHS also inhibited pacemaker amplitude and pacemaker frequency with increases in the resting currents in the outward direction. ODQ, a guanylate cyclase inhibitor, or glibenclamide, an ATP-sensitive K+ channel inhibitor, blocked the SNAP+NaHS-induced inhibition of pacemaker currents in ICC. Also, we found that SNAP+NaHS inhibited the spontaneous intracellular Ca2+ ([Ca2+]i) oscillations in cultured ICC. In conclusion, this study describes the enhanced inhibitory effects of NO plus H2S on ICC in the mouse small intestine. NO+H2S inhibited the pacemaker activity of ICC by modulating intracellular Ca2+. These results may be evidence of a physiological interaction of NO and H2S in ICC for modulating gastrointestinal motility.

The Inhibitory Effects of Hydrogen Sulfide on Pacemaker Activity of Interstitial Cells of Cajal from Mouse Small Intestine

In this study, we studied whether hydrogen sulfide (H2S) has an effect on the pacemaker activity of interstitial cells of Cajal (ICC), in the small intestine of mice. The actions of H2S on pacemaker activity were investigated using whole-cell patch-clamp technique, intracellular Ca2+ analysis at 30degrees C and RT-PCR in cultured mouse intestinal ICC. Exogenously applied sodium hydrogen sulfide (NaHS), a donor of hydrogen sulfide, caused a slight tonic inward current on pacemaker activity in ICC at low concentrations (50 and 100 micrometer), but at high concentration (500 micrometer and 1 mM) it seemed to cause light tonic inward currents and then inhibited pacemaker amplitude and pacemaker frequency, and also an increase in the resting currents in the outward direction. Glibenclamide or other potassium channel blockers (TEA, BaCl2, apamin or 4-aminopydirine) did not have an effect on NaHS-induced action in ICC. The exogenous application of carbonilcyanide p-triflouromethoxyphenylhydrazone (FCCP) and thapsigargin also inhibited the pacemaker activity of ICC as NaHS. Also, we found NaHS inhibited the spontaneous intracellular Ca2+ ([Ca2+]i) oscillations in cultured ICC. In doing an RT-PCR experiment, we found that ICC enriched population lacked mRNA for both CSE and CBS, but was prominently detected in unsorted muscle. In conclusion, H2S inhibited the pacemaker activity of ICC by modulating intracellular Ca2+. These results can serve as evidence of the physiological action of H2S as acting on the ICC in gastrointestinal (GI) motility.

Regional Distribution of Interstitial Cells of Cajal, (ICC) in Human Stomach

We elucidated the distribution of interstitial cells of Cajal (ICC) in human stomach, using cryosection and c-Kit immunohistochemistry to identify c-Kit positive ICC. Before c-Kit staining, we routinely used hematoxylin and eosin (HE) staining to identify every structure of human stomach, from mucosa to longitudinal muscle. HE staining revealed that the fundus greater curvature (GC) had prominent oblique muscle layer, and c-Kit immunostaining c-Kit positive ICC cells were found to have typical morphology of dense fusiform cell body with multiple processes protruding from the central cell body. In particular, we could observe dense processes and ramifications of ICC in myenteric area and longitudinal muscle layer of corpus GC. Interestingly, c-Kit positive ICC-like cells which had morphology very similar to ICC were found in gastric mucosa. We could not find any significant difference in the distribution of ICC between fundus and corpus, except for submucosa where the density of ICC was much higher in gastric fundus than corpus. Furthermore, there was no significant difference in the density of ICC between each area of fundus and corpus, except for muscularis mucosa. Finally, we also found similar distribution of ICC in normal and cancerous tissue obtained from a patient who underwent pancreotomy and gastrectomy. In conclusion, ICC was found ubiquitously in human stomach and the density of ICC was significantly lower in the muscularis mucosa of both fundus/corpus and higher in the submucosa of gastric fundus than corpus.

Voltage-dependent Ca2+ Current Identified in Freshly Isolated Interstitial Cells of Cajal (ICC) of Guinea-pig Stomach

The properties of voltage dependent Ca2+ current (VDCC) were investigated in interstitial cells of Cajal (ICC) distributed in the myenteric layer (ICC-MY) of guinea-pig antrum. In tissue, ICC-MY showed c-Kit positive reactions and produced driving potentials with the amplitude and frequency of about 62 mV and 2 times min(-1), respectively, in the presence of 1micrometer nifedipine. Single ICC-MY isolated by enzyme treatment also showed c-Kit immunohistochemical reactivity. These cells were also identified by generation of spontaneous inward current under K+-rich pipette solution. The voltage clamp experiments revealed the amplitude of - 329 pA inward current at irregular frequency. With Cs+-rich pipette solution at Vh=?80 mV, ICC-MY produced voltage-dependent inward currents (VDIC), and nifedipine (1micrometer) blocked VDIC. Therefore, we successfully isolated c-Kit positive single ICC from guinea-pig stomach, and found that ICC-MY potently produced dihydropiridine sensitive L-type voltage-dependent Ca2+ currents (VDCCL).

(-)-Epigallocatechin Gallate Inhibits the Pacemaker Activity of Interstitial Cells of Cajal of Mouse Small Intestine

The effects of (-)-epigallocatechin gallate (EGCG) on pacemaker activities of cultured interstitial cells of Cajal (ICC) from murine small intestine were investigated using whole-cell patch-clamp technique at 30degrees C and Ca2+ image analysis. ICC generated spontaneous pacemaker currents at a holding potential of -70 mV. The treatment of ICC with EGCG resulted in a dose-dependent decrease in the frequency and amplitude of pacemaker currents. SQ-22536, an adenylate cyclase inhibitor, and ODQ, a guanylate cyclase inhibitor, did not inhibit the effects of EGCG. EGCG-induced effects on pacemaker currents were not inhibited by glibenclamide, an ATP-sensitive K+ channel blocker and TEA, a Ca2+-activated K+ channel blocker. Also, we found that EGCG inhibited the spontaneous [Ca2+]i oscillations in cultured ICC. In conclusion, EGCG inhibited the pacemaker activity of ICC and reduced [Ca2+]i oscillations by cAMP-, cGMP-, ATP-sensitive K+channel-independent manner.

Involvement of Thromboxane A2 in the Modulation of Pacemaker Activity of Interstitial Cells of Cajal of Mouse Intestine

Although many studies show that thromboxane A2 (TXA2) has the action of gastrointestinal (GI) motility using GI muscle cells and tissue, there are no reports on the effects of TXA2 on interstitial cells of Cajal (ICC) that function as pacemaker cells in GI tract. So, we studied the modulation of pacemaker activities by TXA2 in ICC with whole cell patch-clamp technique. Externally applied TXA2 (5 micrometer) produced membrane depolarization in current-clamp mode and increased tonic inward pacemaker currents in voltage-clamp mode. The tonic inward currents by TXA2 were inhibited by intracellular application of GDP-beta-S. The pretreatment of ICC with Ca2+ free solution and thapsigargin, a Ca2+-ATPase inhibitor in endoplasmic reticulum, abolished the generation of pacemaker currents and suppressed the TXA2-induced tonic inward currents. However, chelerythrine or calphostin C, protein kinase C inhibitors, did not block the TXA2-induced effects on pacemaker currents. These results suggest that TXA2 can regulate intestinal motility through the modulation of ICC pacemaker activities. This modulation of pacemaker activities by TXA2 may occur by the activation of G protein and PKC independent pathway via extra and intracellular Ca2+ modulation.

The Morphometric Changes of the Enteric Nervous System in the Intestinal Transection-reanastomosis Model / 체질인류학회지

This study was performed to investigate the morphometric changes of myenteric plexus and type 1 interstitial cells of Cajal (ICC-I) in regeneration process of small intestine transection. Sprague Dawley rats (200~250 g) were anesthetized with ether; then the full thickness of ileal wall were semitransected; and subsequent end-to-end anastomosis were performed by using 6-0 silk suture thread. Sham-operated rats, which only underwent the laparotomy, were used for control group. Experimental animals were sacrificed at 3 days, 7 days, 15 days, and 30 days after the operation. In each group myenteric plexus and ICC-Is were prepared by histochemical method (NADH-TR stain for myenteric plexus, ZIO stain for ICC-Is) and cell numbers were counted by image analyzer (Image plus pro-5.0, Media Cybermedics, USA). Degeneration of myenteric neurons and ICC-Is occurred simultaneously and it was similar in oral and anal to the site of transection. Degeneration effects were most prominent at 3 days and 7 days after intestinal transection. In myenteric plexus, many neurons had degenerated appearances and about 40% of them were lost. Most of ICC-Is had cytoplasmic vacuoles and 20~37% of the cells were lost. At 15 days after transection, there were no more degeneration in myenteric neurons (20~25% cell loss) and ICC-Is (20~13% cell loss). At 30 days after transection, numbers of myenteric neurons were not recovered as that of the control group. However numbers of ICC-Is were as similar as that of the control group. In conclusion, we confirmed that degeneration effects of intestinal transection are more severe in myenteric plexus than in ICC-Is, and recovery of cell loss occurs more slowly in myenteric plexus.

The Effect of External Divalent Cations on Intestinal Pacemaking Activity

Electrical rhythmicity in the gastrointestinal (GI) muscles is generated by pacemaker cells, known as interstitial cells of Cajal (ICC). In the present study, we investigated the effect of external divalent cations on pacemaking activity in cultured ICC from murine small intestine by using whole-cell patch clamp techniques. ICC generated pacemaker currents under a voltage clamp or electrical pacemaker potentials under a current clamp, and showed a mean amplitude of -500+/-50 pA or 30+/-1 mV and the frequency of 18+/-2 cycles/min. Treatments of the cells with external 0 mM Ca2+ stopped pacemaking activity of ICC. In the presence of 2 mM Ca2+, 0 mM external Mg2+ depolarized the resting membrane potential, and there was no change in the frequency of pacemaking activity. However, 10 mM external Mg2+ decreased the frequency of pacemaking activity (6.75+/-1 cycles/min, n=5). We replaced external 2 mM Ca2+ with equimolar Ba2+, Mn2+ and Sr2+, and they all developed inward current in the sequence of Ba2+> Mn2+> Sr2+. Also the frequency of the pacemaking activity was stopped or irregulated. We investigated the effect of 10 mM Ba2+, Mn2+ and Sr2+ on pacemaking activity of ICC in the presence of external 0 mM Mg2+, and found that 10 mM Ba2+ and Mn2+ induced large inward current and stopped the pacemaking activity of ICC (n=5). Interestingly, 10 mM Sr2+ induced small inward current and potentiated the amplitude of pacemaking activity of ICC (n=5). These results indicate that extracellular Ca2+ and Mg2+ are requisite for the pacemaking activity of ICC.

Vanilloid Receptor Type-1 Immunoreactivities in the Mouse Myenteric Plexus: Immunohistochemical and Electrophysiological Study / 체질인류학회지

The vanilloid receptor type-1 (VR1) is a nonselective cation channel activated by capsaicin and can be act as mediator of chemical and physical stimuli that elicit pain. The presence of VR1 in the dorsal root, trigeminal and nodose ganglia has been firmly established, but it unclear in the mouse intestinal wall. The distribution of VR1 receptors in mouse afferent neurons innervating the intestinal tract was investigated by immunohistochemistry. Also small and large intestines were dual-labelled with antibody for VR1 and marker for interstitial cells of Cajal (c-kit). VR1-immunopositive cells were localized on fine fibers in myenteric plexus and expressed weakly myenteric ganglia. The majority of VR1-immunopositive fibers are not colocalized with or apposed to c-kit positive interstitial cells of Cajal. Also electrophysiologically capsaicin had no effect on cultured interstitial cells of Cajal. It is concluded that VR1-immunoreactive intestinal nerves are mainly distributed in myenteric plexus of murine intestinal wall, and vanillod may be not directly related to interstitial cells of Cajal in regulation of intestinal motility.

Vanilloid Receptor Type-1 Immunoreactivities in the Mouse Myenteric Plexus: Immunohistochemical and Electrophysiological Study / 체질인류학회지

The vanilloid receptor type-1 (VR1) is a nonselective cation channel activated by capsaicin and can be act as mediator of chemical and physical stimuli that elicit pain. The presence of VR1 in the dorsal root, trigeminal and nodose ganglia has been firmly established, but it unclear in the mouse intestinal wall. The distribution of VR1 receptors in mouse afferent neurons innervating the intestinal tract was investigated by immunohistochemistry. Also small and large intestines were dual-labelled with antibody for VR1 and marker for interstitial cells of Cajal (c-kit). VR1-immunopositive cells were localized on fine fibers in myenteric plexus and expressed weakly myenteric ganglia. The majority of VR1-immunopositive fibers are not colocalized with or apposed to c-kit positive interstitial cells of Cajal. Also electrophysiologically capsaicin had no effect on cultured interstitial cells of Cajal. It is concluded that VR1-immunoreactive intestinal nerves are mainly distributed in myenteric plexus of murine intestinal wall, and vanillod may be not directly related to interstitial cells of Cajal in regulation of intestinal motility.

Plasticity of Interstitial Cells of Cajal (ICC) and Intestinal Motility in Murine Small Bowel Obstruction / 체질인류학회지

Interstitial cells of Cajal (ICC) are the pacemakers in gastrointestinal slow wave, and also transduce signal inputs from the enteric nervous system to smooth muscle. The abnormal motility corresponded to a lack or decreasing of ICC and a disruption of electrical slow waves. So we developed partial obstruction model in murine small intestine, and found that ICC and electrical slow wave were absent or decreased oral to the occlusion site in previous study. In an additional series of experiments, we examined the ability of tissue regenerate the ICC phenotype and normal electrical slow waves after surgical treatment to relieve the mechanical obstruction, and the animals were allowed to recover for 1~2 months. Removal of the obstruction led to the normal gross appearance and the redevelopment of ICC and recovery of slow wave activity within 30 days. These data demonstrate the plasticity of ICC networks in response to partial obstruction, and suggest that adult tissue retain the ability to regenerate functional ICC. This model may be useful for estimating molecular factors responsible for the regulation of the ICC phenotype. More work is needed to find out the factors in ICC for the therapy of intestinal motility disorders.

The Morphological Study of the Protein Gene Product (PGP) 9.5-like Immunoreactive Enteric Nervous System in Murine Ileum: Close Relation to Interstitial Cells of Cajal (ICC) in Confocal Laser Scanning Microscopy / 대한해부학회지

This study was performed to investigate the morphology of the enteric nervous system and interstitial cells of Cajal (ICC) in the murine ileum. The PGP9.5-like immunoreactive (PGP9.5-LI) neurons and the c-Kit-like immunoreactive (c-Kit-LI) ICCs were stained by indirect immunofluorescence method and were observed under the confocal laser scanning microscopy. According to three dimensional reconstruction study, it was found that the PGP9.5-LI neurons and the c-Kit-LI ICCs were widely distributed in the intestinal wall : (1) In circular muscle layer, PGP9.5-LI nerve fibers were paralell to circular muscle layer. (2) In the myenteric plexus, the PGP9.5-LI nerve were closely apposed to the adjacent PGP9.5-LI nerve, constituting the networks. (3) In double-labeling immunohistochemistry using anti-PGP9.5 and anti-c-Kit antibodies, the c-Kit-LI networks encircled around the neural strands. The characteristic arrangement of the PGP9.5-LI enteric nervous system and the ICC containing c-Kit-positive cells provide a morphological basis upon the mechanism regulating gastro-intestinal motility and the pathogenesis of gatro-intestinal disorders.

Change of Interstitial Cells of Cajal (ICC) and Intestinal Motility in Murine Small Bowel Obstruction / 대한해부학회지

Interstitial cells of Cajal (ICC) are the pacemakers in gastrointestinal slow wave, and also transduce signal inputs from the enteric nervous system to smooth muscle. The abnormal motility corresponded to a lack or decreasing of ICC and a disruption of electrical slow waves. So we developed partial obstruction model in murine small intestine and investigated changes in the ICC networks and electrical activity in the obstructed bowel using c-kit immunohistochemistry and intracelluar electrophysiological techniques. Two weeks following the onset of a partial obstruction, the small intestine increased in diameter and muscular hypertrophy was developed oral to the obstruction site. ICC were absent or only weak at 1 ~25 mm oral to the occlusion site, and this disruption was accompanied by the loss of electrical slow wave. ICC networks and slow waves were normal appearance aboral to the clip. In conclusion, The present results showed that partial intestinal obstruction induced the loss of ICC networks and slow waves. These result will provide a valuable aid for understanding pathogenesis of intestinal motility disorder, and this model may be an important tool for evaluating genetic or molecular factor for the therapeutic opportunities of motility disorder in human.

Morphology of the c-Kit-Immunoreactive Interstitial Cells of Cajal (ICC) in the Mouse Intestine / 대한해부학회지

Interstitial cells of Cajal (ICC) are the pacemalkers in gastrointestinal muscles, and these cells also mediate or transduce inputs from the enteric nervoius system. Immunolabelling of interstitial cells of ICC in intestinal wall is recently developed by using specific marker, anti-c-kit antibody. Immunohistochemistry was done for c-Kit-positive ICC network in attempt to provide a morphological basis for the mechanism regulating gastro-intestinal movement. Cryosection and whole-mount preparations of mouse ileum and colon were immunolabelled using the anti-c-Kit. Immunolabelled specimens were observed under a confocal laser scanning microscopy. According to three dimensional reconstruction study, it was found that the c-Kit-positive cells were widely distributed in the intestinal wall: (1) circular muscle layer, (2) myenteric plexus, (3) deep muscular plexus in ileum, (4) submucosal plexus and longitudinal muscle layer in colon. The characteristic profiles of ICC containing c-Kit-positive cells provide a morphological basis upon the mechanism regulating gastro-intestinal motility.