Related Experiment Video

Updated: Jun 9, 2026

Isolation of Peritoneum-derived Mast Cells and Their Functional Characterization with Ca2+-imaging and Degranulation Assays
11:31

Isolation of Peritoneum-derived Mast Cells and Their Functional Characterization with Ca2+-imaging and Degranulation Assays

Published on: July 4, 2018

P2Y(13) receptor is responsible for ADP-mediated degranulation in RBL-2H3 rat mast cells

Zhan-Guo Gao1, Yi Ding, Kenneth A Jacobson

  • 1Molecular Recognition Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0810, USA. zg21o@nih.gov

Pharmacological Research
|September 4, 2010
PubMed

Insights

Extracellular adenosine diphosphate (ADP) triggers calcium release via the P2Y(1) receptor and hexosaminidase release via the P2Y(13) receptor in rat mast cells. P2Y(13) antagonists may treat allergies.

Area of Science:

  • Immunology
  • Cell Biology
  • Pharmacology

Background:

  • Extracellular adenosine diphosphate (ADP) is a key signaling molecule with diverse physiological functions.
  • Mast cells play a critical role in allergic responses and inflammation.

Purpose of the Study:

  • To identify and characterize the P2Y(13) receptor in rat mast cells (RBL-2H3).
  • To investigate the functional roles of ADP in mast cell activation, specifically calcium mobilization and mediator release.
  • To elucidate the specific purinergic receptors (P2Y(1) and P2Y(13)) involved in ADP-mediated responses.

Main Methods:

  • Gene expression analysis to confirm receptor presence in RBL-2H3 cells.
  • Intracellular calcium mobilization assays using selective agonists and antagonists.
  • Hexosaminidase (Hex) release assays to measure mast cell degranulation.
  • Pharmacological inhibition using phospholipase C inhibitor (U73122) and pertussis toxin (PTX).
  • RNA interference (siRNA) to silence P2Y(1) receptor expression.

Main Results:

  • ADP induced both intracellular calcium mobilization and hexosaminidase (Hex) release in RBL-2H3 cells.
  • Calcium mobilization was mediated by the G(q)-coupled P2Y(1) receptor, as evidenced by sensitivity to U73122 and MRS2500, and insensitivity to PTX.
  • Hex release was primarily mediated by the G(i)-coupled P2Y(13) receptor, indicated by PTX sensitivity and blockade by P2Y(13) antagonist MRS2211.
  • ADP demonstrated higher potency and efficacy in inducing Hex release compared to the P2Y(1) selective agonist MRS2365.
  • RBL-2H3 cells expressed high levels of both P2Y(1) and P2Y(13) receptors.

Conclusions:

  • ADP activates distinct signaling pathways through P2Y(1) and P2Y(13) receptors in rat mast cells.
  • P2Y(1) receptor mediates ADP-induced calcium influx.
  • P2Y(13) receptor mediates ADP-induced hexosaminidase release and enhances antigen-triggered degranulation.
  • Selective P2Y(13) receptor antagonists represent a potential therapeutic strategy for allergic diseases.

More Related Videos

Investigating Mast Cell Secretory Granules; from Biosynthesis to Exocytosis16:01

Investigating Mast Cell Secretory Granules; from Biosynthesis to Exocytosis

Published on: January 26, 2015

Related Concept Videos

IP3/DAG Signaling Pathway01:11

IP3/DAG Signaling Pathway

Membrane lipids such as phosphatidylinositol (PI) are precursors for several membrane-bound and soluble second messengers. Specific kinases phosphorylate PI and produce phosphorylated inositol phospholipids. One such inositol phospholipids are the  phosphatidylinositol-4,5 bisphosphate [PI(4,5)P2], present in the inner half of the lipid bilayer. Upon ligand binding, GPCR stimulates Gq proteins to turn on phospholipase Cꞵ. Activated phospholipase Cꞵ cleaves PI(4,5)P2 and produces two-second...
GPCRs Regulate Adenylyl Cylase Activity01:09

GPCRs Regulate Adenylyl Cylase Activity

Some GPCRs transmit signals through adenylyl cyclase (AC), a transmembrane enzyme. AC helps synthesize second messenger cyclic adenosine monophosphate (cAMP). AC catalyzes cyclization reaction and converts ATP to cAMP by releasing a pyrophosphate. The pyrophosphate is further hydrolyzed to phosphate by the enzyme pyrophosphatase, which drives cAMP synthesis to completion. However, cAMP is rapidly degraded to 5′ AMP by the enzymes phosphodiesterase (PDE), preventing overstimulation of cells.
Two...
Antiplatelet Drugs: Prostaglandin Synthesis, P2Y12 and Glycoprotein IIb/IIIa Inhibitors01:20

Antiplatelet Drugs: Prostaglandin Synthesis, P2Y12 and Glycoprotein IIb/IIIa Inhibitors

Antiplatelet drugs emerge as frontline defenders against the insidious threat of thromboembolic diseases, where abnormal clots obstruct vital blood vessels. These drugs stand as bulwarks, inhibiting platelet aggregation and clot formation, thereby mitigating the risk of life-threatening conditions like myocardial infarction, coronary artery disease, and thrombotic strokes.
Prostaglandin synthesis inhibitors, exemplified by the widely known aspirin, wield their power by irreversibly acetylating...
GPCR Desensitization01:12

GPCR Desensitization

G protein-coupled receptor (GPCR) signaling plays a crucial role in cell functioning. GPCR desensitization is an equally essential process. It allows cells to respond to changing environments and regain sensitivity to new stimuli while preventing unnecessary stimulation when no longer needed. Prolonged exposure to stimuli leads to GPCR desensitization. It involves blocking the receptors from binding and activating additional G proteins. This inhibits activation of downstream effectors, thereby...
Intracellular Signaling Affects Focal Adhesions01:17

Intracellular Signaling Affects Focal Adhesions

Integrins act both as extracellular input receivers and as intracellular processing activators. As their name suggests, integrins are entirely integrated into the membrane structure. Their hydrophobic membrane-spanning regions interact with the phospholipid bilayer's hydrophobic region. These membrane receptors provide extracellular attachment sites for effectors like hormones and growth factors. They activate intracellular response cascades when their effectors are bound and active.
Some...
Adrenergic Receptors: ɑ Subtype01:31

Adrenergic Receptors: ɑ Subtype

Adrenoceptors are classified into α and ꞵ classes based on their potencies to catecholamine agonists. α-adrenoceptors show the following order of catecholamine potency:
Adrenaline ≥ Noradrenaline >> Isoprenaline
α-adrenoceptors are further divided into α1 and α2-adrenoceptors.
α1-Adrenoceptors: These receptors are located postsynaptically on the effector organs and cause constriction of smooth muscle mediated by activation of phospholipase C—inositol-1,4,5-trisphosphate...