一种用于测量器官培养的胚胎鸡十二指肠黏膜表面单向钙内流的简单技术。

A simple technique for the measurement of unidirectional calcium influx at the mucosal surface of organ-cultured embryonic chick duodenum.

作者信息

Corradino R A

机构信息

Department of Physical Biology/Section of Physiology, New York State College of Veterinary Medicine, Cornell University, Ithaca 14853, USA.

出版信息

Anal Biochem. 1978 Nov;91(1):60-9. doi: 10.1016/0003-2697(78)90815-1.

DOI:10.1016/0003-2697(78)90815-1

PMID:9762083

Abstract

Unidirectional calcium influx can be measured by application of a filter paper blotter saturated with a 45Ca-labeled buffer solution to the mucosal surfaces of slit-open embryonic chick duodena which had been cultured mucosal side up on stainless-steel grids. The results obtained with this simple technique support the contention of a direct involvement of a vitamin D-induced calcium-binding protein in the transport of calcium across the mucosal surface. Permeability and inulin space measurements validate the method for measurement of calcium influx at initial velocity and obviate concern for "unstirred layer" effects. Kinetic analyses indicate a two step influx process, one saturable at about 5 mM calcium and the other essentially unsaturable.

摘要

单向钙内流可以通过将用45Ca标记的缓冲溶液浸湿的滤纸吸墨纸应用于 slit-open胚胎鸡十二指肠的粘膜表面来测量，这些十二指肠已在不锈钢网格上粘膜面朝上培养。用这种简单技术获得的结果支持了维生素D诱导的钙结合蛋白直接参与钙跨粘膜表面运输的观点。通透性和菊粉空间测量验证了以初始速度测量钙内流的方法，并消除了对 “未搅动层” 效应的担忧。动力学分析表明存在两步内流过程，一个在约5 mM钙时饱和，另一个基本不饱和。