Purinergic receptors have different effects in rat exocrine pancreas. calcium signals monitored by Fura-2 using confocal microscopy

被引:27
作者
Novak, I [1 ]
Nitschke, R [1 ]
Amstrup, J [1 ]
机构
[1] Univ Copenhagen, August Krogh Inst, DK-2100 Copenhagen O, Denmark
关键词
P2; receptors; ATP; CLSM; luciferin; pancreatic acini; pancreatic ducts;
D O I
10.1159/000063784
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Pancreatic ducts have several types of purinergic P2 receptors, however, nothing is known about P2 receptors in acini. The aim was to establish whether acini express functional P2 receptors coupled to intracellular Ca2+ signals and to measure the signals ratiometrically in a confocal laser scanning microscope. Rat acini and ducts were loaded with various Ca2+ sensing fluorophores (Fluo-4, Fura-Red, Calcium Green-1, Indo-1 and Fura-2). Only Fura-2 loaded evenly into acinar clusters and ducts and UV laser excitation at 351 and 364 nm gave signals showing opposite sensitivity to Ca2+ concentration changes. About 15% of acini or single cells within acini responded to ATP or UTP, yet all responded to carbachol. In contrast, all ducts responded to ATP/UTP. ATP stimulated uptake of a fluorescent anion, luciferin, but again only in a few acinar cells. RT-PCR analysis on acini samples collected from whole pancreas suspensions revealed transcripts for P2Y(2), P2Y(4) and P2X(1), P2X(4) receptors. The low number of functional P2 receptors in acini might be related to the fact that they release ATP. Thereby acini would avoid autocrine stimulation and initiation of autodigestive processes, such as occurs in pancreatitis. Copyright (C) 2002 S. Karger AG, Basel.
引用
收藏
页码:83 / 92
页数:10
相关论文
共 24 条
[1]   An antagonist-insensitive P-2X receptor expressed in epithelia and brain [J].
Buell, G ;
Lewis, C ;
Collo, G ;
North, RA ;
Surprenant, A .
EMBO JOURNAL, 1996, 15 (01) :55-62
[2]   Different purinergic receptors lead to intracellular calcium increases in pancreatic ducts [J].
Christoffersen, BC ;
Hug, MJ ;
Novak, I .
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 1998, 436 (01) :33-39
[3]   Purinoceptors evoke different electrophysiological responses in pancreatic ducts -: P2Y inhibits K+ conductance, and P2X stimulates cation conductance [J].
Hede, SE ;
Amstrup, J ;
Christoffersen, BC ;
Novak, I .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (45) :31784-31791
[4]   Luminal ATP stimulates fluid and HCO3- secretion in guinea-pig pancreatic duct [J].
Ishiguro, H ;
Naruse, X ;
Kitagawa, M ;
Hayakawa, T ;
Case, RM ;
Steward, MC .
JOURNAL OF PHYSIOLOGY-LONDON, 1999, 519 (02) :551-558
[5]   ACTIVATION OF P-2Z PURINOCEPTORS DIMINISHES THE MUSCARINIC CHOLINERGIC-INDUCED RELEASE OF INOSITOL 1,4,5-TRISPHOSPHATE AND STORED CALCIUM IN RAT PAROTID ACINI - ATP AS A COTRANSMITTER IN THE STIMULUS-SECRETION COUPLING [J].
JORGENSEN, TD ;
GROMADA, J ;
TRITSARIS, K ;
NAUNTOFTE, B ;
DISSING, S .
BIOCHEMICAL JOURNAL, 1995, 312 :457-464
[6]   Characterization and localization of P2 receptors in rat submandibular gland acinar and duct cells [J].
Lee, MG ;
Zeng, WZ ;
Muallem, S .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (52) :32951-32955
[7]   Photolysis of caged compounds characterized by ratiometric confocal microscopy: A new approach to homogeneously control and measure the calcium concentration in cardiac myocytes [J].
Lipp, P ;
Luscher, C ;
Niggli, E .
CELL CALCIUM, 1996, 19 (03) :255-266
[8]   A modified confocal laser scanning microscope allows fast ultraviolet ratio imaging of intracellular Ca2+ activity using Fura-2 [J].
Nitschke, R ;
Wilhelm, S ;
Borlinghaus, R ;
Leipziger, J ;
Bindels, R ;
Greger, R .
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 1997, 433 (05) :653-663
[9]   ELECTROPHYSIOLOGICAL STUDY OF TRANSPORT-SYSTEMS IN ISOLATED PERFUSED PANCREATIC DUCTS - PROPERTIES OF THE BASOLATERAL MEMBRANE [J].
NOVAK, I ;
GREGER, R .
PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 1988, 411 (01) :58-68
[10]   A NEW PREPARATION OF PANCREATIC DUCTS FOR PATCH-CLAMP STUDIES [J].
NOVAK, I ;
HUG, MJ .
CELLULAR PHYSIOLOGY AND BIOCHEMISTRY, 1995, 5 (05) :344-352