Different responses of the blockade of the P2Y1 receptor with BPTU in human and porcine intestinal tissues and in cell cultures

• Published in: Neurogastroenterology and motility Vol. 33; no. 7; pp. e14101 - n/a
• Main Authors: Traserra, Sara, Barber, Claudia, Maclnnes, Jane, Relea, Lucia, MacPherson, Lewis C., Cunningham, Margaret R., Vergara, Patri, Accarino, Anna, Kennedy, Charles, Jimenez, Marcel
• Format: Journal Article
• Language: English
• Published: England Wiley Subscription Services, Inc 01.07.2021
• Subjects: Allosteric properties; Animals; BPTU; Calcium (intracellular); Calcium imaging; Cell culture; Cell Culture Techniques; Colon; Gastric motility; Humans; Ileum; Intestines - drug effects; Intestines - metabolism; Mice; Microelectrodes; Motility; Muscle contraction; Muscle, Smooth - drug effects; Neurotransmission; Organ Culture Techniques; P2Y1 receptors; Purinergic P2Y Receptor Antagonists - pharmacology; purinergic response; Receptors, Purinergic P2Y1 - metabolism; Small intestine; Smooth muscle; Swine; tSA201 cells; BPTU; colon; P2Y1 receptors; purinergic response; tSA201 cells
• ISSN: 1350-1925; 1365-2982
• DOI: 10.1111/nmo.14101

Background Gastrointestinal smooth muscle relaxation is accomplished by activation of P2Y1 receptors, therefore this receptor plays an important role in regulation of gut motility. Recently, BPTU was developed as a negative allosteric modulator of the P2Y1 receptor. Accordingly, the aim of this study was to assess the effect of BPTU on purinergic neurotransmission in pig and human gastrointestinal tissues. Methods Ca2+ imaging in tSA201 cells that express the human P2Y1 receptor, organ bath and microelectrodes in tissues were used to evaluate the effects of BPTU on purinergic responses. Key results BPTU concentration dependently (0.1 and 1 µmol L−1) inhibited the rise in intracellular Ca2+ evoked by ADP in tSA201 cells. In the pig small intestine, 30 µmol L−1 BPTU reduced the fast inhibitory junction potential by 80%. Smooth muscle relaxations induced by electrical field stimulation were reduced both in pig ileum (EC50 = 6 µmol L−1) and colon (EC50 = 35 µmol L−1), but high concentrations of BPTU (up to 100 µmol L−1) had no effect on human colonic muscle. MRS2500 (1 µmol L−1) abolished all responses. Finally, 10 µmol L−1 ADPβS inhibited spontaneous motility and this was partially reversed by 30 µmol L−1 BPTU in pig, but not human colonic tissue and abolished by MRS2500 (1 µmol L−1). Conclusions & inferences BPTU blocks purinergic responses elicited via P2Y1 receptors in cell cultures and in pig gastrointestinal tissue. However, the concentrations needed are higher in pig tissue compared to cell cultures and BPTU was ineffective in human colonic tissue. In this study, pharmacological properties of different orthosteric (MRS2179, MRS2279, MRS2500) and allosteric (BPTU) blockers have been tested in human and porcine intestinal tissues and in cell cultures. The picture describes the pathway involved in smooth muscle relaxation.