Fig 1: Adult CNS neuron cultures(A) Neurons from different CNS regions exhibit distinct phenotypic characteristics. Labeling for neuronal marker Tuj1. FezF2 (inset) identifies layer 5b glutamatergic neurons. Cerebellar Purkinje cell neuron labels for calbindin (inset) and spinal cord neuron labels for choline acetyltransferase (ChAT, inset). Neurons isolated at age 6 weeks. Scale bars, 20 µm.(B) Motor cortex neurons form distinct dendritic compartments identified by MAP2 after 5 days in vitro. Tuj1 red, MAP2 green. Neurons isolated at age 6 weeks. Scale bar, 20 µm.(C) Tau labeling (green) identifies axons in culture. At the axon tip is a growth cone. Tuj1 red, with Tau only panel shown below. Neurons isolated at age 4 weeks and cultured for 5 days in vitro. Scale bar, 20 µm.(D) Growth cone (arrow and inset) with lamellipodia is evident in cortical cultures isolated at age 4 weeks, with 7 additional days in vitro. Labeled for Tau. Scale bar, 20 µm.(E) Vesicular glutamate-1 (VGlut1) is present in soma and at bouton-like structures (arrows) indicating excitatory synapses. Tuj1 red. Neurons isolated at age 6 weeks. Scale bars, 10 and 5 µm.(F) Synaptophysin boutons present on motor cortical cell bodies and on Tuj1-labeled neurites. Neurons were isolated at age 6 weeks. Scale bar, 10 µm.(G) Post-synaptic marker Homer (green) localizes with the presynaptic marker synaptophysin (red) at the junction of two Tuj1-labeled processes (blue), indicating a synapse (arrows). Inset with Homer and synaptophysin. Neurons isolated at age 9 weeks. Scale bars, 1 and 2.5 µm.(H) FACS analysis of enriched neuronal fraction. 82.8% of cells are neuronal, 8.6% astrocytes, 3.37% oligodendrocytes, 1.18% microglia, and 2.08% endothelial cells. Flow cytometry antibodies: neurons (Tuj1), astrocytes (ACSA), oligodendrocytes (O4), microglia (CD45, CD11b), and endothelial cells (CD31). Flow cytometry dyes include APC (allophycocyanin), PE (phycoerythrin), VioBlue, VioBright-FITC, and PE-Vio770. Cells isolated at age 8 weeks. Error bars = STDEV.