P2Y2 and P2X4 Receptors Mediate Ca2+ Mobilization in DH82 Canine Macrophage Cells

Purinergic receptors of the P2 subclass are commonly found in human and rodent macrophages where they can be activated by adenosine 5'-triphosphate (ATP) or uridine 5'-triphosphate (UTP) to mediate Ca2+ mobilization, resulting in downstream signalling to promote inflammation and pain. However, little is understood regarding these receptors in canine macrophages. To establish a macrophage model of canine P2 receptor signalling, the expression of these receptors in the DH82 canine macrophage cell line was determined by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry. P2 receptor function in DH82 cells was pharmacologically characterised using nucleotide-induced measurements of Fura-2 AM-bound intracellular Ca2+. RT-PCR revealed predominant expression of P2X4 receptors, while immunocytochemistry confirmed predominant expression of P2Y2 receptors, with low levels of P2X4 receptor expression. ATP and UTP induced robust Ca2+ responses in the absence or presence of extracellular Ca2+. ATP-induced responses were only partially inhibited by the P2X4 receptor antagonists, 2',3'-O-(2,4,6-trinitrophenyl)-ATP (TNP-ATP), paroxetine and 5-BDBD, but were strongly potentiated by ivermectin. UTP-induced responses were near completely inhibited by the P2Y2 receptor antagonists, suramin and AR-C118925. P2Y2 receptor-mediated Ca2+ mobilization was inhibited by U-73122 and 2-aminoethoxydiphenyl borate (2-APB), indicating P2Y2 receptor coupling to the phospholipase C and inositol triphosphate signal transduction pathway. Together this data demonstrates, for the first time, the expression of functional P2 receptors in DH82 canine macrophage cells and identifies a potential cell model for studying macrophage-mediated purinergic signalling in inflammation and pain in dogs.

Medienart:

E-Artikel

Erscheinungsjahr:

2020

Erschienen:

2020

Enthalten in:

Zur Gesamtaufnahme - volume:21

Enthalten in:

International journal of molecular sciences - 21(2020), 22 vom: 13. Nov.

Sprache:

Englisch

Beteiligte Personen:

Sophocleous, Reece Andrew [VerfasserIn]
Miles, Nicole Ashleigh [VerfasserIn]
Ooi, Lezanne [VerfasserIn]
Sluyter, Ronald [VerfasserIn]

Links:

Volltext

Themen:

Calcium
Canine
DH82
Dog
Journal Article
Macrophage
Neuroinflammation
P2X4 receptor
P2Y2 receptor
Pain
Purinergic P2X Receptor Antagonists
Purinergic P2Y Receptor Antagonists
Purinergic signalling
Receptors, Purinergic P2X4
Receptors, Purinergic P2Y2
SY7Q814VUP

Anmerkungen:

Date Completed 03.03.2021

Date Revised 03.03.2021

published: Electronic

Citation Status MEDLINE

doi:

10.3390/ijms21228572

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM317702335