Purinergic 2X 4 (P2X4), but not P2X7, receptors increase cytosolic [Ca2+] and stimulate mucin secretion in rat conjunctival goblet cells to maintain ocular surface health

Fjaervoll, Haakon; Fjaervoll, Ketil; Yang, Menglu; Bair, Jeffrey; Utheim, Tor Paaske; Dartt, Darlene A.

doi:10.1016/j.exer.2023.109614

Experimental Eye Research

2023

DOI: 10.1016/j.exer.2023.109614

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Purinergic 2X 4 (P2X4), but not P2X7, receptors increase cytosolic [Ca2+] and stimulate mucin secretion in rat conjunctival goblet cells to maintain ocular surface health

Haakon Fjaervoll

Ketil Fjaervoll

Menglu Yang

et al.

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2024

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Purinergic agonists increase [Ca²⁺]_i in rat conjunctival goblet cells through ryanodine receptor type 3

Fjaervoll,

Yang

et al. 2024

American Journal of Physiology-Cell Physiology

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ATP and BzATP increase free cytosolic Ca2+ concentration ([Ca2+]i) in conjunctival goblet cells (CGCs) resulting in mucin secretion. The purpose of this study was to investigate the source of the Ca2+i mobilized by ATP and BzATP. First passage cultured rat CGCs were incubated with Fura-2/AM and [Ca2+]i was measured under several conditions with ATP and BzATP stimulation. The following conditions were used: 1) preincubation with the Ca2+ chelator EGTA, 2) preincubation with the SERCA inhibitor thapsigargin (10-6 M) which depletes ER Ca2+ stores, 3) preincubation with phospholipase C (PLC) or protein kinase A (PKA) inhibitor, or 4) preincubation with the voltage-gated calcium channel antagonist nifedipine (10-5 M) and the ryanodine receptor (RyR) antagonist dantrolene (10-5 M). Immunofluorescence microscopy (IF) and RT-qPCR were used to investigate RyR presence in rat and human CGCs. ATP stimulated peak [Ca2+]i was significantly lower after chelating Ca2+i with 2 mM EGTA in Ca2+-free buffer. The peak [Ca2+]i increase in CGCs preincubated with thapsigargin, PKA inhibitor H89, nifedipine and dantrolene, but not the PLC inhibitor, was reduced for ATP at 10-5 M and BzATP at 10-4 M. Incubating CGCs with dantrolene alone decreased [Ca2+]i, and induced CGC cell death at a high concentration. RyR3 was detected in rat and human CGCs with IF and RT-qPCR. We conclude that ATP and BzATP-induced Ca2+i increases originate from the ER, and that RyR3 may be an essential regulator of CGC [Ca2+]i. This study contributes to the understanding of diseases arising from defective Ca2+ signaling in non-excitable cells.

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Purinergic agonists increase [Ca²⁺]_i in rat conjunctival goblet cells through ryanodine receptor type 3

Fjaervoll,

Yang

et al. 2024

American Journal of Physiology-Cell Physiology

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Purinergic 2X 4 (P2X4), but not P2X7, receptors increase cytosolic [Ca2+] and stimulate mucin secretion in rat conjunctival goblet cells to maintain ocular surface health

Cited by 1 publication

References 56 publications

Purinergic agonists increase [Ca²⁺]_i in rat conjunctival goblet cells through ryanodine receptor type 3

Purinergic agonists increase [Ca²⁺]_i in rat conjunctival goblet cells through ryanodine receptor type 3

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Purinergic 2X 4 (P2X4), but not P2X7, receptors increase cytosolic [Ca2+] and stimulate mucin secretion in rat conjunctival goblet cells to maintain ocular surface health

Cited by 1 publication

References 56 publications

Purinergic agonists increase [Ca2+]i in rat conjunctival goblet cells through ryanodine receptor type 3

Purinergic agonists increase [Ca2+]i in rat conjunctival goblet cells through ryanodine receptor type 3

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Purinergic agonists increase [Ca²⁺]_i in rat conjunctival goblet cells through ryanodine receptor type 3

Purinergic agonists increase [Ca²⁺]_i in rat conjunctival goblet cells through ryanodine receptor type 3