The neuronal properties of separate dissociated cell cultures of dorsal and ventral halves of the embryonic mouse spinal cord (E 13.5) were investigated. Ventral-half cultures grew on a variety of substrates and in a variety of media; dorsal-half cultures required a non-neuronal feeder layer and supplemented medium for survival. The two types of cultures differed in their morphological and biochemical properties. Ventral-half neurons remained well separated on the culture plate, whereas dorsal-half neurons tended to aggregate. Lucifer yellow fills showed that ventral-half neurons were substantially larger and had more processes than dorsal-half neurons. Because of the large size and good separation of the neurons, ventral-half cultures provide an especially attractive system for electrophysiologic and morphologic studies. Ventral-half cultures were highly enriched for choline acetyltransferase (ChAT) activity and had more neurons that stained for intracellular acetylcholinesterase (AChE); dorsal-half cultures were enriched for glutamic acid decarboxylase (GAD) activity, and high-affinity gamma-aminobutyric acid (GABA) uptake. The clear differences between the two cultures indicate that many morphological and biochemical properties are already specified on embryonic day 13.5.
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