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. 2012 Feb 23;18(7):526–536. doi: 10.1089/ten.tec.2011.0587

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Copyright 2012, Mary Ann Liebert, Inc.

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FIG. 4. — Assessment of neuronal growth in interface gels. (A) Neuronal growth was investigated using immunostaining and confocal microscopy, for GFP (green), βIII tubulin (red), and Hoechst (blue) at days 5, 10, and 15 in culture. Representative confocal projections are shown of control gels (an interface gel of dissociated GFP DRG cells with dissociated WT DRG cells) and test gels (an interface gel of GFP astrocytes with dissociated WT DRG cells). Arrowheads indicate neurites growing parallel to the interface. Scale bar=150 μm. Neuronal growth was assessed in control gels and test gels at day 5 in culture. Color images available online at www.liebertonline.com/tec

(B) The number of neurites was quantified in confocal projections. While there was a trend for a reduced number of neurites in the DRG:astrocyte interface gels, this did not reach statistical significance (p=0.0547 at the interface vs. control DRG:DRG interfaces). (C) Mean neurite length did not significantly differ between the different interfaces, at or away from the interface. (D) Analysis of the angle of neurite growth revealed differences between the control and test interfaces. Neurites were present at all angles in control interfaces, whereas in test interfaces, neurites were predominantly orientated parallel to the interface with astrocytes. (E) Significantly fewer neurites in the test gels crossed the interface compared with control gels in which more neurites (GFP positive) crossed the interface. ***p<0.001.

FIG. 4. — Assessment of neuronal growth in interface gels. (A) Neuronal growth was investigated using immunostaining and confocal microscopy, for GFP (green), βIII tubulin (red), and Hoechst (blue) at days 5, 10, and 15 in culture. Representative confocal projections are shown of control gels (an interface gel of dissociated GFP DRG cells with dissociated WT DRG cells) and test gels (an interface gel of GFP astrocytes with dissociated WT DRG cells). Arrowheads indicate neurites growing parallel to the interface. Scale bar=150 μm. Neuronal growth was assessed in control gels and test gels at day 5 in culture. Color images available online at www.liebertonline.com/tec

(B) The number of neurites was quantified in confocal projections. While there was a trend for a reduced number of neurites in the DRG:astrocyte interface gels, this did not reach statistical significance (p=0.0547 at the interface vs. control DRG:DRG interfaces). (C) Mean neurite length did not significantly differ between the different interfaces, at or away from the interface. (D) Analysis of the angle of neurite growth revealed differences between the control and test interfaces. Neurites were present at all angles in control interfaces, whereas in test interfaces, neurites were predominantly orientated parallel to the interface with astrocytes. (E) Significantly fewer neurites in the test gels crossed the interface compared with control gels in which more neurites (GFP positive) crossed the interface. ***p<0.001.