Enhanced BDNF signalling following chronic hypoxia potentiates catecholamine release from cultured rat adrenal chromaffin cells
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Key pointsWe investigated the role of the neurotrophin BDNF signalling via the TrkB receptor in rat adrenomedullary chromaffin cells (AMCs) exposed to normoxia (Nox; 21% O2) and chronic hypoxia (CHox; 2% O2) in vitro for ∼48 h.TrkB receptor expression was upregulated in primary AMCs and in immortalized chromaffin (MAH) cells exposed to CHox; this effect was absent in MAH cells deficient in the transcription factor, hypoxia inducible factor (HIF)‐2α.Relative to normoxic controls, activation of the TrkB receptor in chronically hypoxic AMCs led to a marked increase in membrane excitability, intracellular [Ca2+], and catecholamine secretion.The BDNF‐induced rise of intracellular [Ca2+] in CHox cells was sensitive to the selective T‐type Ca2+ channel blocker TTA‐P2 and tetrodotoxin (TTX), suggesting key roles of low threshold T‐type Ca2+ and voltage‐gated Na+ channels in the signalling pathway.AbstractEnvironmental stressors, including chronic hypoxia, enhance the ability of adrenomedullary chromaffin cells (AMCs) to secrete catecholamines; however, the underlying molecular mechanisms remain unclear. Here, we investigated the role of brain‐derived neurotrophic factor (BDNF) signalling in rat AMCs exposed to chronic hypoxia. In rat adrenal glands, BDNF and its tropomyosin‐related kinase B (TrkB) receptor are highly expressed in the cortex and medulla, respectively. Exposure of AMCs to chronic hypoxia (2% O2; 48 h) in vitro caused a significant increase to TrkB mRNA expression. A similar increase was observed in an immortalized chromaffin cell line (MAH cells); however, it was absent in MAH cells deficient in the transcription factor HIF‐2α. A specific TrkB agonist, 7,8‐dihydroxyflavone (7,8‐DHF), stimulated quantal catecholamine secretion from chronically hypoxic (CHox; 2% O2) AMCs to a greater extent than normoxic (Nox; 21% O2) controls. Activation of TrkB by BDNF or 7,8‐DHF increased intracellular Ca2+ ([Ca2+]i), an effect that was significantly larger in CHox cells. The 7,8‐DHF‐induced [Ca2+]i rise was sensitive to the tyrosine kinase inhibitor K252a and nickel (2 mm), but not the Ca2+ store‐depleting agent cyclopiazonic acid. Blockade of T‐type calcium channels with TTA‐P2 (1 μm) or voltage‐gated Na+ channels with TTX inhibited BDNF‐induced [Ca2+]i increases. BDNF also induced a dose‐dependent enhancement of action potential firing in CHox cells. These data demonstrate that during chronic hypoxia, enhancement of BDNF‐TrkB signalling increases voltage‐dependent Ca2+ influx and catecholamine secretion in chromaffin cells, and that T‐type Ca2+ channels play a key role in the signalling pathway.
