Effect of muscarinic agents on sclera fibroblast and their role in myopia

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Effect of muscarinic agents on sclera fibroblast and their role in myopia

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Fig 3. Sclera from myopic eye Sclera from non myopic control eye Fig3A. Chick sclera from experimentally myopic and non myopic control eye. The outer fibrous layer thinned while the inner cartilaginous layer thickened. Atropine receiving myopic eye Atropine receiving non myopic control eye Fig3B. Chick sclera of atropine treated experimentally myopic and control eye. Atropine increased thickness of fibrous layer and decreased cartilaginous layer. The change was towards to a structure of sclera from non myopic eye. Fig 8A. Effect of atropine inhibiting SF cell proliferation in culture. Chick SF were incubated with atropine. Phase contrast photomicrographs were taken after 24 hrs (magnification x100). A. Control culture without atropine. B. Atropine 100 µM caused a significant growth inhibition. The cell morphology appeared to be undisturbed. Fig8B. Effect of atropine on DNA integrity (TUNNEL assay). Chick SF were grown on chamber slides and incubated with atropine (0.1-100 µM). TUNNEL assay was performed after 24 hrs and photomicrographs are taken (magnification x100). Positive control (control culture stimulated to undergo apotosis produced dark brown stain in apoptic nuclei) B. Atropine 0.1 µM C. Atropine µM D. Atropine 10 µM E. Atropine 100 µM Fig10A. Immumocytochemistry of mAchR subtypes in human SF in culture. Subtype selective antibodies bound to cultured SF demonstrates the presence of m1, m2, m4 receptors (shown in green). When secondary FITC labeled antibody was used without the primary antibody no significant binding was observed (negative control). m1 m4 m2 negative control Fig10B. Protein corresponding to mAchR subtypes m1,m2,m4, and m5 identified in mouse sclera by immunoblotting. Subtype selective antibodies bound to specific protein bands in mouse sclera extract. Sepcific bands did not appear in the control experiments, in which the primary antibody is omitted. Numbers on left of panels are molecular mass markers (Kd). M1 Negative control M2 M4 M5 Fig 21A. Chick sclera protein extract and SF cell lysates were analysed by gelatin zymography. Lane and 2; chick sclera fibroblast cell lysate, Lane and 4; chick sclera protein extract Fig19B. Westernblot ananlysis of chick sclera protein extract and SF cell lysate using antihuman MMP-2 antibody. Lane ;MMP-2 standard, Lane ; chick sclera fibroblast cell lysate Lane ; chick sclera protein extract Fig19C. Western blot analysis of chick sclera protein extract and SF cell lysate using anti human MMP-9 antibody. Lane 1: chick sclera fibroblast cell lysate Lane 2: chick sclera protein extract Land 3: MMP-9 standard Diagram 1. Mechanism of growth factor action and related events Diagram 2. Interaction between muscarinic and EGF receptors in growth regulation. Diagram 3. Mechanism of G protein coupled m1 receptor. . Atropine increased thickness of fibrous layer and decreased cartilaginous layer. The change was towards to a structure of sclera from non myopic eye. Fig 8A. Effect of atropine inhibiting. cartilaginous layer thickened. Atropine receiving myopic eye Atropine receiving non myopic control eye Fig3B. Chick sclera of atropine treated experimentally myopic and control. growth inhibition. The cell morphology appeared to be undisturbed. Fig8B. Effect of atropine on DNA integrity (TUNNEL assay). Chick SF were grown on chamber slides and incubated with atropine

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