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981.
982.
J Riedlberger C Amsler M Doser U Straumann P Tru?l D Bailey S Barlag U Gastaldi R Landua C Sabev KD Duch M Heel H Kalinowsky F Kayser E Klempt B May O Schreiber P Weidenauer M Ziegler W Dahme F Feld-Dahme U Schaefer WR Wodrich S Ahmad JC Bizot B Delcourt J Jeanjean H Nguyen N Prevot EG Auld DA Axen KL Erdman B Howard R Howard BL White M Comyn G Beer GM Marshall LP Robertson M Botlo C Laa H Vonach 《Canadian Metallurgical Quarterly》1989,40(6):2717-2731
983.
Plasma levels of tissue plasminogen activator (tPA) increase rapidly in response to specific vasoactive agents, trauma, and neural stimulation. This response has been attributed to acute release of tPA from stored pools within the vascular endothelium and from catecholamine storage vesicles of chromaffin cells. We have tested directly whether tPA can be sorted into the regulated secretory pathway using the murine pituitary-derived neuroendocrine cell line AtT-20 transfected with tPA cDNA. Clones of AtT-20 cells expressing tPA were isolated, and targeting of tPA into the regulated secretory pathway was demonstrated by (1) stimulation of tPA secretion with 8-bromo-cAMP, the secretagogue which promotes the release of dense granule contents; (2) colocalization with ACTH, an endogenous protein that is stored in dense core granules; and (3) retention of newly synthesized tPA in the cell for prolonged periods of time. Laser scanning confocal microscopy analysis of cells immunostained with antibodies to tPA and ACTH showed colocalization at the tips of the neuritic processes under the cytoplasmic membrane, a region where dense granules are known to migrate after maturation. Treatment of the cells with 5 mM 8-bromo-cAMP for 30 min resulted in a 2.41+/-0.36-fold increase in tPA secretion. Both the magnitude of the stimulatory effect and the fraction of the intracellular tPA released were the same regardless of the tPA expression level in the various clones. Pulse-chase experiments showed that a portion of newly synthesized tPA is retained in the cell for at least 4 h and is released into the culture medium in response to 8-bromo-cAMP. These studies indicate that tPA, under the appropriate conditions, can be targeted into the regulated secretory pathway and can be stored for later release by cellular stimuli. 相似文献
984.