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A and B, leptin receptor immunoreactivity on neuronal somata, illustrating punctate labelling within the cytoplasm (A) and labelling associated with the plasma membrane (B). A is a Z-projection of a series of confocal images taken at 1 μm intervals, B is a single confocal section. Primary antibody labelling was visualised with an Alexa 488-conjugated donkey anti-goat secondary antibody (green). C, D, E and F, merged, single plane confocal images comparing leptin receptor labelling with monoclonal markers (red; Cy3) for soma/dendrites (MAP2; C), axons (GAP43, D) or synaptic terminals (synapsin 1; E and F). The yellow areas correspond to points of overlap. In these images, the confocal gain and/or laser intensity was increased to facilitate the visualisation of fine processes. Leptin receptor immunoreactivity was associated with synaptic markers in older cultures (9-14 days, E and F). In the zoomed image (F), arrowheads show examples of co-localisation with <t>synapsin</t> <t>1</t> at individual puncta. Strong leptin receptor immunoreactivity was also associated with dendritic (C) and axonal (D and E) growth cones (arrows). Scale bars are 10 μm (A, B, D, E and F) and 20 μm (C).
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A and B, leptin receptor immunoreactivity on neuronal somata, illustrating punctate labelling within the cytoplasm (A) and labelling associated with the plasma membrane (B). A is a Z-projection of a series of confocal images taken at 1 μm intervals, B is a single confocal section. Primary antibody labelling was visualised with an Alexa 488-conjugated donkey anti-goat secondary antibody (green). C, D, E and F, merged, single plane confocal images comparing leptin receptor labelling with monoclonal markers (red; Cy3) for soma/dendrites (MAP2; C), axons (GAP43, D) or synaptic terminals (synapsin 1; E and F). The yellow areas correspond to points of overlap. In these images, the confocal gain and/or laser intensity was increased to facilitate the visualisation of fine processes. Leptin receptor immunoreactivity was associated with synaptic markers in older cultures (9-14 days, E and F). In the zoomed image (F), arrowheads show examples of co-localisation with synapsin 1 at individual puncta. Strong leptin receptor immunoreactivity was also associated with dendritic (C) and axonal (D and E) growth cones (arrows). Scale bars are 10 μm (A, B, D, E and F) and 20 μm (C).

Journal:

Article Title: Leptin inhibits epileptiform-like activity in rat hippocampal neurones via PI 3-kinase-driven activation of BK channels

doi: 10.1113/jphysiol.2002.029488

Figure Lengend Snippet: A and B, leptin receptor immunoreactivity on neuronal somata, illustrating punctate labelling within the cytoplasm (A) and labelling associated with the plasma membrane (B). A is a Z-projection of a series of confocal images taken at 1 μm intervals, B is a single confocal section. Primary antibody labelling was visualised with an Alexa 488-conjugated donkey anti-goat secondary antibody (green). C, D, E and F, merged, single plane confocal images comparing leptin receptor labelling with monoclonal markers (red; Cy3) for soma/dendrites (MAP2; C), axons (GAP43, D) or synaptic terminals (synapsin 1; E and F). The yellow areas correspond to points of overlap. In these images, the confocal gain and/or laser intensity was increased to facilitate the visualisation of fine processes. Leptin receptor immunoreactivity was associated with synaptic markers in older cultures (9-14 days, E and F). In the zoomed image (F), arrowheads show examples of co-localisation with synapsin 1 at individual puncta. Strong leptin receptor immunoreactivity was also associated with dendritic (C) and axonal (D and E) growth cones (arrows). Scale bars are 10 μm (A, B, D, E and F) and 20 μm (C).

Article Snippet: In dual labelling experiments, monoclonal markers for MAP2, GAP43 and synapsin 1 (all from Sigma) were visualised with a Cy3-conjugated donkey anti-mouse secondary antibody (Jackson ImmunoResearch).

Techniques: