Inhibitory neurons from fetal rat cerebral cortex exert delayed axon formation and active migration in vitro

Abstract

Inhibitory and excitatory neurons exhibit distinct patterns of development in the mammalian cerebral cortex. The morphological development of inhibitory and excitatory neurons derived from fetal rat cerebral cortex has now been compared in vitro. Inhibitory neurons were identified by immunofluorescence staining with antibodies to γ-aminobutyric acid, and axon formation was detected by staining with antibodies to phosphorylated neurofilaments. In chemically defined, glia-free and low-density cultures, excitatory neurons formed axons within three days of plating. By contrast, inhibitory neurons required more than six days to form axons. Time-lapse analysis over six days revealed that most inhibitory neurons were bipolar and that their two processes exhibited alternate growth and retraction without giving rise to axons. Movement of the cell body towards the growing process was apparent in about one-half of inhibitory neurons, whereas such movement was never seen in excitatory neurons. The migratory behavior of neurons was further investigated by culture on a glial cell monolayer. Inhibitory neurons migrated over substantially larger distances than did excitatory neurons. The centrosome of inhibitory neurons translocated to the base of the newly emerging leading process, suggesting the existence of a force that pulls intracellular organelles towards the leading process. Centrosome translocation was not detected in excitatory neurons. These observations suggest that the developmental programs of excitatory and inhibitory neurons differ. Inhibitory neurons thus possess a more effective cytoskeletal machinery for migration than excitatory neurons and they form axons later.