Related Experiment Videos
P2-purinoceptors utilize multiple signalling pathways in MDCK-D1 cells
P A Insel1, B L Firestein, M Xing
1Dept of Pharmacology, University of California, San Diego, La Jolla 92093, USA.
Journal of Autonomic Pharmacology
|December 1, 1996
Summary
Madin-Darby canine kidney (MDCK)-D1 cells express both P2a and P2y purinoceptors, activating phospholipase A2 and leading to arachidonic acid release. These receptors also influence other signaling pathways, impacting cell function.
Area of Science:
- Cell Biology
- Molecular Pharmacology
- Epithelial Cell Biology
Background:
- Madin-Darby canine kidney (MDCK) cells are a standard model for epithelial cell research.
- MDCK-D1 cells, a clonal variant, were used to investigate purinergic signaling.
Purpose of the Study:
- To determine the subtypes of P2-purinoceptors expressed in MDCK-D1 cells.
- To elucidate the downstream signaling pathways activated by these purinoceptors.
Main Methods:
- Analysis of arachidonic acid release in response to various nucleotide analogues.
- Use of specific antagonists (suramin) and agonists (2-methylthio ATP, UTP) to differentiate receptor activity.
- Measurement of inositol trisphosphate formation, phosphatidylcholine hydrolysis, and phospholipase D activity.
- Assessment of cyclic AMP (cAMP) levels and the role of cyclooxygenase.
Main Results:
- MDCK-D1 cells co-express P2a and P2y-purinoceptors.
- Both receptor subtypes are linked to arachidonic acid release via phospholipase A2 activation.
- ATP and UTP differentially affect inositol trisphosphate formation and phosphatidylcholine hydrolysis.
- Purine nucleotides stimulate cAMP production, partly through cyclooxygenase activation and prostaglandin E2 release.
Conclusions:
- MDCK-D1 cells possess co-expressed P2a and P2y-purinoceptors.
- These receptors regulate cell function through multiple mechanisms, including phospholipase activation.
- Nucleotide signaling involves autocrine/paracrine actions of signaling molecules like PGE2.