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内质网蛋白对小肠Cajal间质细胞内钙的调节作用

Regulation of Intracellular Calcium by Endoplasmic Reticulum Proteins in Small Intestinal Interstitial Cells of Cajal.

作者信息

Park Chan Guk, Wu Mei Jin, Hong Chansik, Jo Ju Yeon, Jiao Han Yi, Park Hyun, Jun Jae Yeoul, Choi Seok

机构信息

Department of Internal Medicine, College of Medicine, Chosun University, Gwangju, Korea.

Department of Medicine, Graduate School, Chosun University, Gwangju, Korea.

出版信息

J Neurogastroenterol Motil. 2018 Jan 30;24(1):128-137. doi: 10.5056/jnm16212.

DOI:10.5056/jnm16212
PMID:28774158
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5753911/
Abstract

BACKGROUND/AIMS: We investigated the role of representative endoplasmic reticulum proteins, stromal interaction molecule 1 (STIM1), and store-operated calcium entry-associated regulatory factor (SARAF) in pacemaker activity in cultured interstitial cells of Cajal (ICCs) isolated from mouse small intestine.

METHODS

The whole-cell patch clamp technique applied for intracellular calcium ions ([Ca]) analysis with STIM1 or SARAF overexpressed cultured ICCs from mouse small intestine.

RESULTS

In the current-clamping mode, cultured ICCs displayed spontaneous pacemaker potentials. External carbachol exposure produced tonic membrane depolarization in the current-clamp mode, which recovered within a few seconds into normal pacemaker potentials. In STIM1-overexpressing cultured ICCs pacemaker potential frequency was increased, and in SARAF-overexpressing ICCs pacemaker potential frequency was strongly inhibited. The application of gadolinium (a non-selective cation channel inhibitor) or a Ca-free solution to understand Orai channel involvement abolished the generation of pacemaker potentials. When recording intracellular Ca concentration with Fluo 3-AM, STIM1-overexpressing ICCs showed an increased number of spontaneous intracellular Ca oscillations. However, SARAF-overexpressing ICCs showed fewer spontaneous intracellular Ca oscillations.

CONCLUSION

Endoplasmic reticulum proteins modulated the frequency of pacemaker activity in ICCs, and levels of STIM1 and SARAF may determine slow wave patterns in the gastrointestinal tract.

摘要

背景/目的：我们研究了内质网代表性蛋白——基质相互作用分子1（STIM1）和储存操纵性钙内流相关调节因子（SARAF）在从小鼠小肠分离的培养Cajal间质细胞（ICC）起搏活动中的作用。

方法

采用全细胞膜片钳技术，对过表达STIM1或SARAF的从小鼠小肠分离的培养ICC进行细胞内钙离子（[Ca]）分析。

结果

在电流钳模式下，培养的ICC呈现出自发性起搏电位。在电流钳模式下，外源性卡巴胆碱暴露导致膜出现强直性去极化，数秒内恢复为正常起搏电位。在过表达STIM1的培养ICC中，起搏电位频率增加，而过表达SARAF的ICC中，起搏电位频率受到强烈抑制。应用钆（一种非选择性阳离子通道抑制剂）或无钙溶液以了解Orai通道的参与情况，结果发现这消除了起搏电位的产生。当用Fluo 3 - AM记录细胞内钙浓度时，过表达STIM1的ICC显示出自发性细胞内钙振荡数量增加。然而，过表达SARAF的ICC显示出自发性细胞内钙振荡较少。

结论

内质网蛋白调节ICC起搏活动的频率，STIM1和SARAF的水平可能决定胃肠道的慢波模式。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f44/5753911/cffe79a0d0d9/jnm-24-128f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f44/5753911/05f622d98f37/jnm-24-128f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f44/5753911/a92e02466f1d/jnm-24-128f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f44/5753911/6f5af264b3f1/jnm-24-128f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f44/5753911/2cb1e0b0a55b/jnm-24-128f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f44/5753911/cffe79a0d0d9/jnm-24-128f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f44/5753911/05f622d98f37/jnm-24-128f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f44/5753911/a92e02466f1d/jnm-24-128f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f44/5753911/6f5af264b3f1/jnm-24-128f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f44/5753911/2cb1e0b0a55b/jnm-24-128f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f44/5753911/cffe79a0d0d9/jnm-24-128f5.jpg

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