Secretory proteins enter the Golgi apparatus when transport vesicles fuse with the cis-side and exit in transport vesicles budding from the trans-side. Resident Golgi enzymes that have been transported in the cis-to-trans direction with the secretory flow must be recycled constantly by retrograde transport in the opposite direction. In this study, we describe the functional characterization of Golgi-derived transport vesicles that were isolated from tissue culture cells. We found that under the steady-state conditions of a living cell, a fraction of resident Golgi enzymes was found in vesicles that could be separated from cisternal membranes. These vesicles appeared to be depleted of secretory cargo. They were capable of binding to and fusion with isolated Golgi membranes, and after fusion their enzymatic contents most efficiently processed cargo that had just entered the Golgi apparatus. Those results indicate a possible role for these structures in recycling of Golgi enzymes in the Golgi stack.
Oculartissues have the capacity to metabolize arachidonate to prostaglandins and related materials, such as hydroxy arachidonate derivatives (HETEs) which are potent mediators of the inflammatory response. Reactive oxygen species are also present during the inflammatory response, resulting in an altered oxidative environment within the eye.This study was designed to evaluate the possible impact of the oxidant hydrogen peroxide and anti-oxidants, ascorbic acid and glutathione, upon arachidonate metabolism. It was found that hydrogen peroxide was observed to potently inhibit arachidonate metabolism in the cornea, but not in the iris-ciliary body. This might be related to a more efficient detoxification of hydrogen peroxide by iris-ciliary body. Ascorbate appeared to have a general stimulatory influence upon arachidonate metabolism in the iris-ciliary body. In the cornea, ascorbate selectively reduced metabolism to HETE while enhancing the products generated by the cyclo-oxygenase pathway. In both cornea and iris-ciliary body reduced glutathione suppressed conversion of arachidonate to its active metabolites.These observations indicate that arachidonate metabolism in ocular tissues is sensitive to the oxidative environment.
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