Abstract EN:

An increase in circulating catecholamine levels is one of the mechanisms whereby organisms cope with stress. In the periphery, catecholamines mainly originate from the sympathoadrenal system. The secretion of catecholamines by adrenal chromaffin cells is a key event in response to stressors and it is chiefly controlled by trans-synaptically released acetylcholine from the splanchnic nerve endings. As supported by earlier results obtained in the laboratory, in addition to the central control through cholinergic innervation, a local gap junction-delineated route mediating intercellular electrical coupling between chromaffin cells is involved in the hormonal secretory process and represents an efficient complement to synaptic transmission able to amplify catecholamine release after synaptic stimulation. Whether these two communication pathways (i. E. Synaptic neurotransmission and gap junctional coupling) contribute to stress-evoked increased catecholamine secretion still remains unknown. We addressed this issue using acute adrenal slices from stressed rats (5 day-cold exposure). Our results show that in cold exposed rats, gap junctional communication undergoes a functional plasticity, as evidenced by an increased number of dye-coupled cells. Of a physiological interest is that this up-regulation results in the appearance of a robust electrical coupling between chromaffin cells that allows the transmission of action potentials between coupled cells. This enhancement of gap junctional communication parallels an increase in expression levels of connexin36 and connexin43 proteins. Both transcriptional and post-translational mechanisms are involved since Cx36 transcripts are increased in stressed rats and the expression of the scaffolding protein Zonula Occludens-1, known to interact with both Cx36 and Cx43, is also up-regulated. Consistent with an up-regulated coupling in stressed rats, the cytosolic Ca2+ concentration ([Ca2+]i) rise triggered in a single cell by an iontophoretic application of nicotine occurs simultaneously in several neighboring cells. We also showed that in response to stress, both chromaffin cell excitability and chemical transmission at the splanchnic nerve terminal-chromaffin cell synapses are increased. We next investigated whether vasopressin (VP), a neuropeptide known to play a key role in stress response, could be involved in increased gap junctional communication in stressed rats. Our results show that VP and d[Leu4,Lys8]VP, a V1b receptor specific agonist, increase the occurrence of gap junction-mediated synchronized [Ca2+]i transients between chromaffin cells, both in stressed and unstressed rats. This suggests that, although VP can up-regulate gap junctional coupling, it is unlikely the main factor involved in increased gap junctional communication observed in response to stress. Exposure to cold also enhances the synaptic neurotransmission between splanchnic nerve endings and chromaffin cells, as evidenced by an increase in spontaneous excitatory post-synaptic currents (sEPSCs). This correlates with an increased density of nerve fibers innervating the medulla. To go further, we examined whether the nAChR subtype formed by the combination of alpha9 and alpha10 subunits recently identified in isolated rat chromaffin cells is involved in this effect. By using a toxin (alpha-RgIa) to specifically block alpha9/alpha10 nAChRs, we showed that alpha9 /alpha10 nAChRs contribute to synaptic transmission. Interestingly, the expression level of alpha9 receptors is up-regulated in cold exposed rats. In addition, we show that in stressed rats, alpha9 /alpha10 nAChRs mainly contribute to acetylcholine-induced currents, as compared to alpha3 nAChRs that is the main nicotinic receptor activated in response to acetylcholine in control rats. This indicates that stress also induces nAChR plasticity, at least by acting on the expression level. In sum, the functional changes occurring both on gap junctional communication and on synaptic transmission converge to improve the stimulus-secretion coupling efficiency in the adrenal gland and may represent endogenous mechanisms by which the adrenal medullary tissue ensures appropriate increased catecholamine secretion in response to stressors