cAMP activates an ATP-conductive pathway in cultured shark rectal gland cells

被引:12
作者
Cantiello, HF
Jackson, GR
Prat, AG
Gazley, JL
Forrest, JN
Ausiello, DA
机构
[1] HARVARD UNIV, SCH MED, DEPT MED, CHARLESTOWN, MA 02129 USA
[2] AMHERST COLL, AMHERST, MA 01002 USA
[3] MT DESERT ISL BIOL LAB, SALSBURY COVE, ME 04672 USA
[4] YALE UNIV, SCH MED, DEPT MED, NEW HAVEN, CT 06520 USA
来源
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY | 1997年 / 272卷 / 02期
关键词
adenosine; 3'; 5'-cyclic monophosphate; adenosine 5'-triphosphate-binding cassette transporters; adenosine 5'-triphosphate release; chloride secretion; purinergic receptors;
D O I
10.1152/ajpcell.1997.272.2.C466
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The molecular mechanisms associated with ATP transport and release into the extracellular milieu are largely unknown. To assess the presence of endogenous ATP-conductive pathway(s) in shark rectal gland (SRG) cells, patch-clamp techniques were applied to primary cultures of SRG cells. Whole cell currents were obtained with either intracellular tris(hydroxymethyl)-aminomethane (Tris) or Mg2+ salts of ATP (200 mM nominal ATP) and 280 mM NaCl bathing solution. Basal currents showed a sizable ATP permeability for outward movement of MgATP. Adenosine 3',5'-cyclic monophosphate (cAMP) stimulation significantly increased the whole cell conductance (with either intracellular Tris-ATP or MgATP). Symmetrical whole cell ATP currents were also observed after cAMP activation, thus consistent with ATP as the main charge carrier. The cAMP-inducible ATP currents were insensitive to the Cl- channel blockers 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid, diphenylamine-2-carboxylate, and anthracene-9-carboxylic acid but were readily blocked by nifedipine (400 mu M) and glibenclamide (400 mu M). The nature of the electrodiffusional ATP movement was further assessed by single-channel analysis of either MgATP or Tris-ATP currents ill excised inside-out patches, both spontaneous and after activation with protein kinase A. Single-channel ATP currents were inhibited by either nifedipine or glibenclamide. Thus SRG cells express endogenous ATP-permeable pathways both before and after cAMP stimulation. Electrodiffusional ATP movement by SRG cells may play a significant role in the transport and delivery of cellular ATP to the extracellular milieu, which may help coordinate the dynamics of the epithelial secretory response in this cell model.
引用
收藏
页码:C466 / C475
页数:10
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