- 作者: Jaehwa Choi; Satin G. Sawant; David B. Couch; Ing Kang Ho; Jerry M. Farley
- 作者服務機構: Department of Pharmacology and Toxicology, University of Mississippi Medical Center, Jackson, Miss., USA
- 中文摘要: --
- 英文摘要: Mitogen- and isoproterenol-induced changes of [Ca2+]i in T cells attached to a glass substrate were examined. Murine (C57BL/6) splenic T cells were attached to coverslips or 35-mm dishes (MatTek) precoated with Cell Tak? (3.5 μg/cm2). The cells were then loaded with fluorescent dye (2 μg/ml of fura2-AM or fluo3-AM) and changes in [Ca2+]i in a population of cells (using a spectrofiuorometer) or in single cells (using a confocal microscope) were measured during continuous superfusion. Population measurements of [Ca2+]i demonstrated that concanavalin A (Con A, 2 or 5 μg/ml) caused an increase in [Ca2+]i that rose to a peak and then declined to a steady state. The concentration-response relationship (0.05-5 μg/ml) had an EC50 of ~0.3 μg/ml. Isoproterenol decreased the Con A-induced elevation of steady state [Ca2+]i. In single cell studies, the increase in [Ca2+]i in response to Con A typically occurred in about 50% of the cells in a microscope field, and the delay before activation varied among cells. Taken together, these data demonstrate that Cell Tak? can be used to attach T cells to glass coverslips and will be useful for the study of signaling mechanisms in T cells.
- 中文關鍵字: --
- 英文關鍵字: T cell ; Cell Tak?; Calcium ; Superfusion ; Spectrofluorometer ; Confocal microscopy ; Concanavalin A ; Isoproterenol