The effect of phospholipase A2 on chloride transport by pancreatic secretory granules.

Secretory granules from the rat pancreas contain electrolyte transport pathways that may contribute to exocrine fluid production. The Cl- selective transport pathway was measured indirectly in isolated granules by ionophore-induced lysis after suspension in isotonic KCl. This Cl- transport was shown to respond to alterations in the granule membrane lipid environment. Exogenously added phospholipase A2 (PLA2) caused an increase of up to 193% in the Cl- specific transport by the isolated granules. In addition, the products of PLA2 hydrolysis, lysophospholipids and unesterified fatty acids, directly increased the rate of Cl- transport when incubated with granules in vitro. Lysophospholipids (2.0 micrograms/ml) increased the Cl- transport between 280-450% (depending on the lysophospholipid species). Similarly, free fatty acids (10 microM) increased the granule Cl- transport from 25% with capric acid (10:0) to 255% with arachidonic acid (20:4). The relative extent of stimulation by fatty acids was dependent on their carbon chain length and to a lesser extent, the degree of unsaturation. The inhibition (68%) of PLA2 promoted granule lysis by 4-acetamido-4'-isothiocyanatostilbene 2,2'-disulfonic acid (0.5 mM) also suggests that the effect is specific for the granule Cl- channel. Thus, the data show that zymogen granule Cl- transport is influenced by membrane lipids and supports a role for PLA2 in controlling electrolyte transport during stimulus-secretion coupling.