FGF-1 Triggers Pannexin-1 Hemichannel Opening in Spinal Astrocytes of Rodents and Promotes Inflammatory Responses in Acute Spinal Cord Slices.
We show here that the growth factor FGF-1 is proinflammatory in the spinal cord and explore the inflammatory mechanisms. FGF-1 applied to rat spinal astrocytes in culture initiates calcium signaling and induces secretion of ATP that within minutes increases membrane permeability to ethidium (Etd(+)) and Ca(2+) by activating P2X7 receptors (P2X7Rs) that open pannexin hemichannels (Px1 HCs) that release further ATP; by 7 h treatment, connexin 43 hemichannels (Cx43 HCs) are also opened. In acute mouse spinal cord slices ex vivo, we found that FGF-1 treatment for 1 h increases the percentage of GFAP-positive astrocytes that show enhanced Px1 HC-mediated Etd(+) uptake. This response to FGF-1 was not observed in astrocytes in slices of cerebral cortex. FGF-1-induced dye uptake by astrocytes is prevented by BAPTA-AM or a phospholipase C (PLC) inhibitor. Furthermore, in spinal cord slices, P2X7R antagonists (BBG and A740003) and Px1 HC blockers ((10)Panx1 and carbenoxolone) prevent the increase in Etd(+) uptake by astrocytes, whereas Gap19, a selective Cx43 HC blocker, has no effect on dye uptake at this time. Microglia are not required for the increase in Etd(+) uptake by astrocytes induced by FGF-1, although they are activated by FGF-1 treatment. The morphological signs of microglia activation are inhibited by P2X7R antagonists and (10)Panx1 and are associated with elevated levels of proinflammatory cytokines in cord slices treated with FGF-1. The FGF-1 initiated cascade may play an important role in spinal cord inflammation in vivo



We find that FGF-1 elevates [Ca(2+)]i in spinal astrocytes, which causes vesicular release of ATP and activation of P2X7Rs to trigger opening of Px1 HCs, which release further ATP. This regenerative response occurs in astrocyte cultures and in acute spinal cord slices. In the latter, FGF-1 application promotes the activation of microglia and increases the production of proinflammatory cytokines through mechanisms depending on P2X7 receptors and Px1 HCs. This proinflammatory microenvironment may favor recruitment of leukocytes into the spinal cord and impacts negatively on neuronal structure and function in vivo Any step in these processes provides a potential therapeutic target for treatment of secondary damage in various spinal cord pathologies.
Department of Anesthesiology, Perioperative Care and Pain Medicine, New York University School of Medicine, New York, New York 10016, Dominick P. Purpura, Department of Neuroscience, Albert Einstein College of Medicine, Bronx, New York 10461, and Juan.GarreCastro@nyumc.org jmgarre@gmail.com michael.bennett@einstein.yu.edu.
Juan Mauricio
JM
Garré
Department of Anesthesiology, Perioperative Care and Pain Medicine, New York University School of Medicine, New York, New York 10016.
Guang
G
Yang
Dominick P. Purpura, Department of Neuroscience, Albert Einstein College of Medicine, Bronx, New York 10461, and Institute of Cardiology, Lithuanian University of Health Sciences, Kaunas LT-50161, Lithuania.
Feliksas F
0000-0002-8172-7804
FF
Bukauskas
Dominick P. Purpura, Department of Neuroscience, Albert Einstein College of Medicine, Bronx, New York 10461, and Juan.GarreCastro@nyumc.org jmgarre@gmail.com michael.bennett@einstein.yu.edu.
Michael V L
MV
Bennett
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism*; D020013:Calcium Signaling / Q000502:physiology*; D002463:Cell Membrane Permeability; D002540:Cerebral Cortex / Q000166:cytology; D018031:Connexin 43 / Q000378:metabolism; D017630:Connexins; D005260:Female; D016220:Fibroblast Growth Factor 1 / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D008297:Male; D051379:Mice; D017628:Microglia / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000502:physiology; D051381:Rats; D013116:Spinal Cord / Q000166:cytology*
D018031:Connexin 43; D017630:Connexins; D009419:Nerve Tissue Proteins; C503631:Panx1 protein, mouse; D016220:Fibroblast Growth Factor 1; D000255:Adenosine Triphosphate
astrocyte; fibroblast growth factor; glia; inflammation; microglia; spinal cord