Differentiation of Apical and Basal Dendrites in Pyramidal Cells and Granule Cells in Dissociated Hippocampal Cultures
Fig 1
The morphology of pyramidal and granule cells at DIV 13 in dissociated culture.
(A) Typical morphology of pyramidal cells, granule cells and interneurons. The cell type was characterized by immunostaining with Math2, Prox1 (magenta) and GAD67, respectively. Pyramidal and granule cells were visualized by GFP transfection. Interneurons were visualized by immunostaining with GAD67. Scale bar: 40 μm. (B) Histogram distribution of somal size in pyramidal and granule cells of DIV 10–13. n = 40 for each cell type. (C) Axon/dendrite specification as demarcated by Ankyrin G expression. Cells were immunostained with Ankyrin G (magenta) and Math2 or Prox1 (blue) at DIV 7. Asterisks indicate axon. Scale bar: 20 μm. (D) Expression of synaptic markers at DIV 16. Boxed regions in the left panel are shown in the right panels. PSD95 (red) was localized in protrusions in both principal (upper) and minor (lower) dendrites. Some PSD95 signals in dendritic protrusions (asterisks) were apposed to Synapsin I signals (green). Dendrites were visualized by GFP expression. Scale bar: 20 μm (left), 2 μm (right). (E-G) Quantification of total dendritic length (E), the number of primary dendrites (F) and branch terminals (G). n = 20. Student's t-test: *P<0.05, **P<0.03, ***P<0.001. (H) Ratio of length of each dendrite arbor to the longest dendrite. All dendrite arbors in a neuron were ranked from the longest to the shortest. n = 20.