Vertical bias in dendritic trees of non-pyramidal neocortical neurons expressing GAD67-GFP in Vitro

The neocortical neuropil has a strong vertical (orthogonal to pia) orientation, constraining the intracortical flow of information and farming the basis for the functional parcellation of the cortex into semi-independent vertical columns or 'modules'. Apical dendrites of excitatory pyramidal neurons are a major component of this Vertical neuropil, but the extent to which inhibitory, GABAergic neurons conform to this structural and functional design is less well documented. We used a gene gun to transfect organotypic slice cultures of meese and rat neocortex with the enhanced green fluorescent protein (eGFP) gene driven by the promoter for glutamic acid decarboxytase 67 (GAD67), an enzyme expressed exclusively in GABAergic cells. Many GAD67-GFP expressing cells were highly fluorescent; and their dendritic morphologies and axonal patterns, revealed in minute detail, were characteristic of GABAergic neurons. We traced 150 GFP-expressing neurons from confocal image stacks, and estimated the degree of vertical bias in their dendritic trees using a novel computational metric. Over 70% of the neurons in our sample had dendritic trees with a highly significant vertical bias. We conclude that GABAergic neurons make an important contribution to the vertical neocortical neuropil, and are likely to integrate synaptic inputs from axons terminating within their own module.