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Little is known about the relationship between stimulation of lacrimal gland LG tear protein secretion by parasympathetic versus sympathetic nerves particularly whether the spectrum of tear proteins evoked through each innervation pathway varies We have previously shown that activity and abundance of cathepsin S CTSS a cysteine protease is greatly increased in tears of Sj gren s syndrome SS patients and in tears from the male NOD mouse of autoimmune dacryoadenitis that recapitulates SS-associated dry eye disease Beyond the increased synthesis of CTSS detected in the diseased NOD mouse LG increased tear CTSS secretion in NOD mouse tears was recently linked to increased exocytosis from a novel endolysosomal secretory pathway Here we have compared secretion and trafficking of CTSS in healthy mouse LG acinar cells stimulated with either the parasympathetic acetylcholine receptor agonist carbachol CCh or the sympathetic 1-adrenergic agonist phenylephrine PE In situ secretion studies show that PE significantly increases CTSS activity and protein in tears relative to CCh stimulation by 1 2-fold p 0 0009 and 5-fold p-0 0319 respectively A similar significant increase in CTSS activity with PE relative to CCh is observed when cultured LGAC are stimulated in vitro CCh stimulation significantly elevates intracellular Ca2 an effect associated with increases in the size of Rab3D-enriched vesicles consistent with compound fusion and subsequently decreases in their intensity of labeling consistent with their exocytosis PE stimulation induces a lower Ca2 response and has minimal effects on Rab3D-enriched SV diameter or the intensity of Rab3D-enriched SV labeling LG deficient in Rab3D exhibit a higher sensitivity to PE stimulation and secrete more CTSS activity Significant increases in the colocalization of endolysosomal vesicle markers Lamp1 Lamp2 Rab7 with the subapical actin suggestive of fusion of endolysosomal vesicles at the apical membrane occur both with CCh and PE stimulation but PE demonstrates increased colocalization In conclusion the 1-adrenergic agonist PE increases CTSS secretion into tears through a pathway independent of the exocytosis of Rab3D-enriched mature SV possibly representing an alternative endolysosomal secretory pathway Copyright 2021 Elsevier Ltd All rights reserved

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Phenylephrine increases tear cathepsin S secretion in healthy murine lacrimal gland acinar cells through an alternative ...

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with Lacrimal gland acinar cells as keyword