Escolar G, White J G
Hospital Clinico y Provincial, Barcelona, Spain.
Blood Cells. 1991;17(3):467-85; discussion 486-95.
Channels of the surface-connected, open canalicular system (OCS) of human platelets serve as the pathway for transport of substances into the cells and as conduits for the discharge of alpha granule products secreted during the platelet release reaction. The purpose of the present study was to determine if both functions of the OCS can take place simultaneously. Suspensions of washed platelets were exposed to thrombin at 1 U/ml for 5, 60, or 180 seconds in the presence of fibrinogen molecules coupled to particles of colloidal gold (Fgn/Au). The samples were fixed in a low concentration of glutaraldehyde and embedded in L.R. White resin to preserve antigenicity. Thin sections were exposed to a rabbit polyclonal antibody to human fibrinogen followed by an anti-rabbit IgG coupled to 5-nm gold beads. Thrombin caused Fgn/Au particles to bind to platelets and enter channels of the surface-connected OCS. Endogenous fibrinogen detected by immunogold 5-nm beads were localized to alpha granules in resting platelets and 5 seconds after thrombin stimulation. At 60 seconds and 3 minutes Fgn/Au particles were present in swollen alpha granules, as well as OCS channels. Fibrinogen gold beads were evident in alpha granules and OCS channels connected to the platelet surface. The 18- to 20-nm Fgn/Au particles were in the same channels of the OCS as fibrinogen gold beads. The OCS is a final common pathway for uptake of particulates and discharge of secretory products in thrombin-activated human platelets.
人血小板表面连接的开放小管系统(OCS)的通道是物质进入细胞的运输途径,也是血小板释放反应过程中分泌的α颗粒产物排出的管道。本研究的目的是确定OCS的这两种功能是否能同时发生。在与胶体金颗粒(Fgn/Au)偶联的纤维蛋白原分子存在的情况下,将洗涤后的血小板悬浮液暴露于1 U/ml的凝血酶中5、60或180秒。样品用低浓度戊二醛固定,包埋在L.R. White树脂中以保留抗原性。将薄片暴露于抗人纤维蛋白原的兔多克隆抗体,然后是与5纳米金珠偶联的抗兔IgG。凝血酶导致Fgn/Au颗粒与血小板结合并进入表面连接的OCS通道。通过免疫金5纳米珠检测到的内源性纤维蛋白原定位于静息血小板的α颗粒以及凝血酶刺激后5秒。在60秒和3分钟时,Fgn/Au颗粒存在于肿胀的α颗粒以及OCS通道中。纤维蛋白原金珠在与血小板表面相连的α颗粒和OCS通道中很明显。18至20纳米的Fgn/Au颗粒与纤维蛋白原金珠位于OCS的相同通道中。OCS是凝血酶激活的人血小板摄取颗粒和分泌产物排出的最终共同途径。