A Role of c-kit tyrosinekinase of physiological development of the gastrointestinal pacemaker activity

c-kit酪氨酸激酶对胃肠起搏器活性生理发育的作用

• 批准号: 06454163
• 负责人: NISHI Katsuhide
• 金额: $ 4.54万
• 依托单位: KUMAMOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06454163/
• 关键词: interstitial cell (IC); pacemaker activity; gastrointestinal tract; c-kit tyrosinekinase; antibody for kit-protein (ACK2); GI motility ileum; cultured c-kit (+) cell; c-kit発現細胞; 抗c-kitモノクローナル抗体(ACK2); 胃平滑筋; 細胞内Ca^<2+>濃度測定; Ca^<2+>-activated K^+

As to origin of electrical and mechanical rhythmicity in the gastrointestinal tract, a class of specialized cells, often referred to as interstitial cells (IC) have been suggested to serve as pacemaker cells However, the function of ICs has been difficult to investigate in intact muscles because of the structural complexities of GI muscle and the IC network. We took a different approach to explore function of the ICs in the intestine. Results are as follows, 1) A certain type of cells in smooth muscle layrs of developing gastrointestinal tract in the mouse expresses Kit-gene which is a proto-oncogene encoding a receptor tyrosine in PDGF/CSF-I receptor family and in vitro injection of a neutralizing antibody (ACK2) for the proto-oncogene product, Kit protein, during the first few days after birth reduced the number of the Kit-expressing cells, which was accompanied by impairment of development of normal rhythmic mechanical activity in the mouse intestine, 2) ACK2 specifically labels int … More erstitial cells (ICs) in the mouse GI tracts, and ACK2-treatment induces a loss of electrical slow wave activity and depolarization of the membranes of smooth muscle cells, 3) we have succeeded in isolating ICs in primary culture of the mice small intestine and recorded spontaneously active electrical currents from these primary cultured ICs, 4) Both of the voltage-sensitive Na^+ and Ca^<2+> currents were not observed clearly in the all c-kit ^+ cells tested, but the rhythmic fluctuation of membrane voltage was observed under the current-clamp condition, where only Cl^- permeates, 5) In the ileum of neonates on the day of birth (day 0 pp) which did not show any spontaneous contractile activity, ACh (>10^<-8>M) induced a dose-related sustained contraction, but the contractile responses to ACh declined in sensitivity each day after birth in the control mice, while ileum of ACK2-treated animals did not show a decrease in the sensitivity to ACh as well as prostaglandin E1 and F2a. These observations indicate that the kit--signaling pathway is important in the development of ICs as pacemaker cells in the intestinal tract. Impairment of development of the IC network would not only affect GI motility but also affect pharmacological characteristics of the ileac smooth muscle. Less

关于胃肠道电节律和机械节律的起源，一类特化细胞，通常被称为间质细胞（IC），被认为是起起搏作用的细胞。然而，由于GI肌肉和IC网络结构的复杂性，IC的功能一直难以在完整肌肉中研究。我们采取了不同的方法来探索IC在肠道中的功能。结果如下：1）小鼠发育中的胃肠道平滑肌层中的某种类型的细胞表达Kit基因，该基因是编码PDGF/CSF-1受体家族中的受体酪氨酸的原癌基因，并且在出生后的最初几天内体外注射针对原癌基因产物Kit蛋白的中和抗体（ACK 2）减少了表达Kit的细胞的数量，伴随着小鼠肠道正常节律性机械活动的发育受损，2）ACK 2特异性标记intr ...更多信息 小鼠胃肠道中的肠上皮细胞（IC），并且ACK 2处理诱导电慢波活性的丧失和平滑肌细胞膜的去极化，3）我们已经成功地在小鼠小肠的原代培养物中分离IC并且记录来自这些原代培养的IC的自发活性电流，（4）所有c-kit ^+细胞均未观察到电压敏感性Na^+和Ca^2+电流，但在电流钳条件下，仅Cl^-渗透时，可观察到膜电压的节律性波动。（5）在新生小鼠的回肠中，ACh（&gt;10 μ M）在出生当天（第0天pp）<-8>引起了一种与剂量相关的持续收缩，但对照小鼠对ACh的收缩反应在出生后每天都在下降，而ACK 2处理动物的回肠没有显示出对ACh以及前列腺素E1和F2 a的敏感性降低。这些观察结果表明，kit-信号通路是重要的，在发展中国家的集成电路作为起搏细胞在肠道。IC网络的发育受损不仅会影响GI运动，而且会影响回肠平滑肌的药理学特性。少

项目成果

Tokutomi N: "Rhythmic Cl-current and physiological roles of the intestinal c-kit-positive cells." Pflugers Arch-Eur.J.Physiol. 431. 169-177 (1995)

Tokutomi N：“肠道 c-kit 阳性细胞的节律性 Cl 电流和生理作用。”

Shigeko Torihashi.: "c-kit-depenpent development of intestitial cells and electrical activity in the murine gastrointestinal tract." Cell Tissue Res. 280. 97-111 (1995)

Shigeko Torihashi.：“小鼠胃肠道中肠细胞的 c-kit 依赖性发育和电活动。”

Yoshiko Tokutomi: "Property the Ca^<2+>-induced Ca^<2+> releasing system in gastric smooth muscle cells anti-c-kit monoclonal antibody (ACK2)-treated mice." Japanese Journal of Pharmacology. 64(S1). 159P (1994)

Yoshiko Tokutomi：“在胃平滑肌细胞抗c-kit单克隆抗体(ACK2)处理的小鼠中确定Ca ^ 2 诱导的Ca ^ 2 释放系统的特性。”

Frank C: "Different effects of baclofen and GTPγS on voltage-activated Ca^<2+> currents in rat hippocampal neurons in vitro." Eur.J.Pharmacol. 295. 87-92 (1996)

Frank C：“巴氯芬和 GTPγS 对体外大鼠海马神经元电压激活 Ca^2+ 电流的不同影响。”Eur.J.Pharmacol。295. 87-92 (1996)

Frank, C: "Different effects of baclofen and GTPgammaS on voltage-activated Ca^<2+> currents in rat hippocampal neurons in vitro" Eur.J.Pharmacol.295. 87-92 (1996)

Frank, C：“巴氯芬和 GTPgammaS 对体外大鼠海马神经元电压激活 Ca^2 > 电流的不同影响”Eur.J.Pharmacol.295。