Mechanosensitive release of adenosine 5′-triphosphate through pannexin channels and mechanosensitive upregulation of pannexin channels in optic nerve head astrocytes: A mechanism for purinergic involvement in chronic strain

• Published in: Glia 2014-09, Vol.62 (9), p.1486-1501
• Main Authors: Beckel, Jonathan M., Argall, Arthur J., Lim, Jason C., Xia, Jingsheng, Lu, Wennan, Coffey, Erin E., Macarak, Edward J., Shahidullah, Mohammed, Delamere, Nicholas A., Zode, Gulab S., Sheffield, Val C., Shestopalov, Valery I., Laties, Alan M., Mitchell, Claire H.
• Format: Article
• Language: English
• Subjects: Adenosine triphosphatase; Adenosine Triphosphate - metabolism; Animals; Astrocytes - drug effects; Astrocytes - physiology; ATP release; cell swelling; Cells, Cultured; Connexins - genetics; Connexins - metabolism; Cytoskeletal Proteins - genetics; Cytoskeletal Proteins - metabolism; Disease Models, Animal; Eye Proteins - genetics; Eye Proteins - metabolism; Female; Glaucoma; Glaucoma - physiopathology; Glycoproteins - genetics; Glycoproteins - metabolism; Male; mechanosensitive signaling; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; Nucleotide Transport Proteins - metabolism; Optic Disk - drug effects; Optic Disk - physiology; Optic nerve; Osmotic Pressure - physiology; P2X7 receptor; pannexin; Purinergic P2X Receptor Antagonists - pharmacology; Rats, Long-Evans; Stress, Mechanical
• ISSN: 0894-1491; 1098-1136
• DOI: 10.1002/glia.22695

As adenosine 5′‐triphosphate (ATP) released from astrocytes can modulate many neural signaling systems, the triggers and pathways for this ATP release are important. Here, the ability of mechanical strain to trigger ATP release through pannexin channels and the effects of sustained strain on pannexin expression were examined in rat optic nerve head astrocytes. Astrocytes released ATP when subjected to 5% of equibiaxial strain or to hypotonic swelling. Although astrocytes expressed mRNA for pannexins 1–3, connexin 43, and VNUT, pharmacological analysis suggested a predominant role for pannexins in mechanosensitive ATP release, with Rho kinase contribution. Astrocytes from panx1−/− mice had reduced baseline and stimulated levels of extracellular ATP, confirming the role for pannexins. Swelling astrocytes triggered a regulatory volume decrease that was inhibited by apyrase or probenecid. The swelling‐induced rise in calcium was inhibited by P2X7 receptor antagonists A438079 and AZ10606120, in addition to apyrase and carbenoxolone. Extended stretch of astrocytes in vitro upregulated expression of panx1 and panx2 mRNA. A similar upregulation was observed in vivo in optic nerve head tissue from the Tg‐MYOCY437H mouse model of chronic glaucoma; genes for panx1, panx2, and panx3 were increased, whereas immunohistochemistry confirmed increased expression of pannexin 1 protein. In summary, astrocytes released ATP in response to mechanical strain, with pannexin 1 the predominant efflux pathway. Sustained strain upregulated pannexins in vitro and in vivo. Together, these findings provide a mechanism by which extracellular ATP remains elevated under chronic mechanical strain, as found in the optic nerve head of patients with glaucoma. GLIA 2014;62:1486–1501 Main Points: Mechanical strain triggers ATP release from astrocytes via pannexin 1. Released ATP autostimulates P2X7 receptors. Sustained stretch increased pannexin expression. Mechanism for sustained elevation of extracellular ATP under chronic stretch.