fusion-activated ca2+ entry an “active zone” of elevated ca2+ during the postfusion stage of lamellar body exocytosis in rat type ii pneumocytesfusion-activated ca2 +进入高架ca2 +的u201c活动区域u201d在postfusion阶段ii型pneumocytes片状体胞外分泌的老鼠.pdfVIP

Fusion-Activated Ca2+ Entry: An ‘‘Active Zone’’ of Elevated Ca2+ during the Postfusion Stage of Lamellar Body Exocytosis in Rat Type II Pneumocytes 1. 1,2. 1 2 1 Pika Miklavc , Manfred Frick , Oliver H. Wittekindt , Thomas Haller , Paul Dietl * 1 Institute of General Physiology, University of Ulm, Ulm, Germany, 2 Department of Physiology and Medical Physics, Medical University of Innsbruck, Innsbruck, Austria Abstract Background: Ca2+ is essential for vesicle fusion with the plasma membrane in virtually all types of regulated exocytoses. However, in contrast to the well-known effects of a high cytoplasmic Ca2+ concentration ([Ca2+] ) in the prefusion phase, the c occurrence and significance of Ca2+ signals in the postfusion phase have not been described before. Methodology/Principal Findings: We studied isolated rat alveolar type II cells using previously developed imaging techniques. These cells release pulmonary surfactant, a complex of lipids and proteins, from secretory vesicles (lamellar bodies) in an exceptionally slow, Ca2+- and actin-dependent process. Measurements of fusion pore formation by darkfield scattered light intensity decrease or FM 1-43 fluorescence intensity increase were combined with analysis of [Ca2+]c by ratiometric Fura-2 or Fluo-4 fluorescence measurements. We found that the majority of single lamellar body fusion events were followed by a transient (t1/2 of decay = 3.2 s) rise of localized [Ca2+]c originating at the site of lamellar body fusion. [Ca2+]c increase f